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亀岡 正典大学院保健学研究科 保健学専攻教授
研究者基本情報
■ 学位■ 研究ニュース
- 2021年04月28日, 「ドライフォグを用いた次亜塩素酸水溶液・過酸化水素水の噴霧による新型コロナウイルス不活性化実験」の産学連携プロジェクト
- 2020年09月10日, 「新型コロナウイルス感染症（COVID-19）に対する経口ワクチンの開発」がAMEDに採択されました
- 2020年09月04日, 新型コロナウイルスに対する安全な創薬スクリーニング系の構築
- 2019年09月27日, インドネシアに流行するHIV-1(エイズウイルス）サブタイプB亜種の侵入経路を推定
- 2018年05月18日, エイズウイルスの調節遺伝子を標的とするゲノム編集法でウイルス遺伝子を不活化
■ 研究分野
研究活動情報
■ 論文- Human immunodeficiency virus type 1 (HIV-1) remains a serious health threat in Indonesia. In particular, the CRF01_AE viruses were the predominant HIV-1 strains in various cities in Indonesia. However, information on the dynamic transmission characteristics and spatial-temporal transmission of HIV-1 CRF01_AE in Indonesia is limited. Therefore, the present study examined the spatial-temporal transmission networks and evolutionary characteristics of HIV-1 CRF01_AE in Indonesia. To clarify the epidemiological connection between CRF01_AE outbreaks in Indonesia and the rest of the world, we performed phylogenetic studies on nearly full genomes of CRF01_AE viruses isolated in Indonesia. Our results showed that five epidemic clades, namely, IDN clades 1-5, of CRF01_AE were found in Indonesia. To determine the potential source and mode of transmission of CRF01_AE, we performed Bayesian analysis and built maximum clade credibility trees for each clade. Our study revealed that CRF01_AE viruses were commonly introduced into Indonesia from Southeast Asia, particularly Thailand. The CRF01_AE viruses might have spread through major pandemics in Asian countries, such as China, Vietnam, and Laos, rather than being introduced directly from Africa in the early 1980s. This study has major implications for public health practice and policy development in Indonesia. The contributions of this study include understanding the dynamics of HIV-1 transmission that is important for the implementation of HIV disease control and prevention strategies in Indonesia.2024年05月, Scientific reports, 14(1) (1), 9917 - 9917, 英語, 国際誌研究論文（学術雑誌）
- 2024年04月, Cells, 英語[査読有り]研究論文（学術雑誌）
- The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), has necessitated the global development of countermeasures since its outbreak. However, current therapeutics and vaccines to stop the pandemic are insufficient and this is mainly because of the emergence of resistant variants, which requires the urgent development of new countermeasures, such as antiviral drugs. Replicons, self-replicating RNAs that do not produce virions, are a promising system for this purpose because they safely recreate viral replication, enabling antiviral screening in biosafety level (BSL)-2 facilities. We herein constructed three pCC2Fos-based RNA replicons lacking some open reading frames (ORF) of SARS-CoV-2: the Δorf2-8, Δorf2.4, and Δorf2 replicons, and validated their replication in Huh-7 cells. The functionalities of the Δorf2-8 and Δorf2.4 replicons for antiviral drug screening were also confirmed. We conducted puromycin selection following the construction of the Δorf2.4-puro replicon by inserting a puromycin-resistant gene into the Δorf2.4 replicon. We observed the more sustained replication of the Δorf2.4-puro replicon by puromycin pressure. The present results will contribute to the establishment of a safe and useful replicon system for analyzing SARS-CoV-2 replication mechanisms as well as the development of novel antiviral drugs in BSL-2 facilities.2023年07月, Virus research, 334, 199176 - 199176, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Hepatitis B virus (HBV) is a leading cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. Current therapeutic drugs for chronic HBV infection use IFN and nucleos(t)ide analogs; however, their efficacy is limited. Thus, there is an urgent need to develop new antivirals for HBV therapy. In this study, we identified a plant-derived polyphenolic bioflavonoid, amentoflavone, as a new anti-HBV compound. Amentoflavone treatment dose-dependently inhibited HBV infection in HBV-susceptible cells with HepG2-hNTCP-C4 and primary human hepatocyte PXB-cells. A mode-of-action study showed that amentoflavone inhibits the viral entry step, but not the viral internalization and early replication processes. Attachment of HBV particles as well as HBV preS1 peptide to HepG2-hNTCP-C4 cells was inhibited by amentoflavone. The transporter assay revealed that amentoflavone partly inhibits uptake of sodium taurocholate cotransporting polypeptide (NTCP)-mediated bile acid. Furthermore, effect of various amentoflavone analogs on HBs and HBe production from HBV-infected HepG2-hNTCP-C4 cells was examined. Robustaflavone exhibited comparable anti-HBV activity to that of amentoflavone and an amentoflavone-7,4', 4‴-trimethyl ether derivative (sciadopitysin) with moderate anti-HBV activity. Cupressuflavone or the monomeric flavonoid apigenin did not exhibit the antiviral activity. Amentoflavone and its structurally related biflavonoids may provide a potential drug scaffold in the design of a new anti-HBV drug inhibitor targeting NTCP.2023年06月, Microbiology and immunology, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- BACKGROUND: Biomarkers that are cost-effective and accurate for predicting severe coronavirus disease 2019 (COVID-19) are urgently needed. We would like to assess the role of various inflammatory biomarkers on admission as disease severity predictors and determine the optimal cut-off of the neutrophile-to-lymphocyte ratio (NLR) for predicting severe COVID-19. METHODS: We conducted a cross-sectional study in six hospitals in Bali and recruited real-time PCR-confirmed COVID-19 patients aged >18 y from June to August 2020. Data collection included each patient's demographic, clinical, disease severity and hematological data. Multivariate and receiver operating characteristic curve analyses were performed. RESULTS: A total of 95 Indonesian COVID-19 patients were included. The highest NLR among severe patients was 11.5±6.2, followed by the non-severe group at 3.3±2.8. The lowest NLR was found in the asymptomatic group (1.9±1.1). The CD4+ and CD8+ values were lowest in the critical and severe disease groups. The area under the curve of NLR was 0.959. Therefore, the optimal NLR cut-off value for predicting severe COVID-19 was ≥3.55, with sensitivity at 90.9% and a specificity of 16.7%. CONCLUSIONS: Lower CD4+ and CD8+ and higher NLR values on admission are reliable predictors of severe COVID-19 among Indonesian people. NLR cut-off ≥3.55 is the optimal value for predicting severe COVID-19.2023年04月, Transactions of the Royal Society of Tropical Medicine and Hygiene, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- 2023年03月, Bulletin of Health Sciences Kobe, 38, 29 - 43, 英語Viability and acceptability of self-sampling as a primary screening method for sexually transmitted infections in female Kobe University students.[査読有り]研究論文（学術雑誌）
- Human immunodeficiency virus type 1 (HIV-1) is characterized by a large degree of genetic variability because of high rates of recombination and mutation, sizable population sizes, and rapid replication. Therefore, the present study investigated HIV-1 subtype distribution and the appearance of drug resistance mutations (DRMs) in viruses that are prevalent in Makassar, South Sulawesi, Indonesia. The HIV-1 pol, env, and gag genes were amplified from 63 infected individuals and sequenced for a subtyping analysis. CRF01_AE was identified as the predominant HIV-1 circulating recombinant form (CRF) in Makassar, South Sulawesi, Indonesia. Subtype B and recombinant viruses containing CRF01_AE, CRF02_AG, and/or subtype B gene fragments were also detected. Several major DRMs against non-nucleoside reverse transcriptase inhibitors were found among antiretroviral therapy (ART)-experienced subjects, while ART-naive subjects did not possess any transmitted drug resistance. The prevalence of DRMs was very high among ART-experienced subjects; therefore, further surveillance is required in this region.2023年01月, AIDS research and human retroviruses, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Chronic hepatitis C virus (HCV) infection can lead to liver cirrhosis and hepatocellular carcinoma. Although current medications using direct-acting antivirals (DAAs) are highly effective and well-tolerated for treating patients with chronic HCV, high prices and the existence of DAA-resistant variants hamper treatment. There is thus a need for easily accessible antivirals with different mechanisms of action. During the screening of Indonesian medicinal plants for anti-HCV activity, we found that a crude extract of Dryobalanops aromatica leaves possessed strong antiviral activity against HCV. Bioassay-guided purification identified an oligostilbene, vaticanol B, as an active compound responsible for the anti-HCV activity. Vaticanol B inhibited HCV infection in a dose-dependent manner with 50% effective and cytotoxic concentrations of 3.6 and 559.5 µg/mL, respectively (Selectivity Index: 155.4). A time-of-addition study revealed that the infectivity of HCV virions was largely lost upon vaticanol B pretreatment. Also, the addition of vaticanol B following viral entry slightly but significantly suppressed HCV replication and HCV protein expression in HCV-infected and a subgenomic HCV replicon cells. Thus, the results clearly demonstrated that vaticanol B acted mainly on the viral entry step, while acting weakly on the post-entry step as well. Furthermore, co-treatment of the HCV NS5A inhibitor daclatasvir with vaticanol B increased the anti-HCV effect. Collectively, the present study has identified a plant-derived oligostilbene, vaticanol B, as a novel anti-HCV compound.2023年, Biological & pharmaceutical bulletin, 46(8) (8), 1079 - 1087, 英語, 国内誌[査読有り]研究論文（学術雑誌）
- Infection with viruses belonging to the genus Flavivirus, such as Japanese encephalitis virus (JEV) and dengue virus (DENV), is a worldwide health problem. Vaccines against JEV and DENV are currently available. However, the dengue vaccine possibly increases the risk of severe dengue due to antibody-dependent enhancement (ADE). Moreover, the Japanese encephalitis (JE) vaccine reportedly induces cross-reactive ADE-prone antibodies against DENV, potentially leading to symptomatic dengue. Therefore, it is necessary to eliminate the risk of ADE through vaccination. In this study, we attempted to develop a JE vaccine that does not induce ADE of DENV infection using an epitope modification strategy. We found that an ADE-prone monoclonal antibody cross-reactive to DENV and JEV recognizes the 106th amino acid residue of the E protein of JEV (E-106). The JE DNA vaccine with a mutation at E-106 (E-106 vaccine) induced comparable neutralizing antibody titers against JEV to those induced by the wild-type JE DNA vaccine. Meanwhile, the E-106 vaccine induced 64-fold less cross-reactive ADE-prone antibodies against DENV. The mutation did not compromise the protective efficacy of the vaccine in the lethal JEV challenge experiment. Altogether, the modification of a single amino acid residue identified in this study helped in the development of an ADE-free JE vaccine.2022年08月, Vaccines, 10(9) (9), 英語, 国際誌[査読有り]研究論文（学術雑誌）
- The increase in antibiotic resistance in non-typhoidal Salmonella enterica (NTS) has been confirmed in Indonesia by this study. We confirmed the virulence genes and antimicrobial susceptibilities of clinical NTS (n = 50) isolated from chicken meat in Indonesia and also detected antimicrobial resistance genes. Of 50 strains, 30 (60%) were non-susceptible to nalidixic acid (NA) and all of them had amino acid mutations in gyrA. Among 27 tetracycline (TC) non-susceptible strains, 22 (81.5%) had tetA and/or tetB. The non-susceptibility rates to ampicillin, gentamicin or kanamycin were lower than that of NA or TC, but the prevalence of blaTEM or aadA was high. Non-susceptible strains showed a high prevalence of virulence genes compared with the susceptible strains (tcfA, p = 0.014; cdtB, p < 0.001; sfbA, p < 0.001; fimA, p = 0.002). S. Schwarzengrund was the most prevalent serotype (23 strains, 46%) and the most frequently detected as multi-antimicrobial resistant. The prevalence of virulence genes in S. Schwarzengrund was significantly higher than other serotypes in hlyE (p = 0.011) and phoP/Q (p = 0.011) in addition to the genes above. In conclusion, NTS strains isolated from Indonesian chicken had a high resistance to antibiotics and many virulence factors. In particular, S. Schwarzengrund strains were most frequently detected as multi-antimicrobial resistant and had a high prevalence of virulence genes.2022年05月, Pathogens (Basel, Switzerland), 11(5) (5), 英語, 国際誌[査読有り]研究論文（学術雑誌）
- The host transmembrane protein MARCH8 is a RING finger E3 ubiquitin ligase that downregulates various host transmembrane proteins, such as MHC-II. We have recently reported that MARCH8 expression in virus-producing cells impairs viral infectivity by reducing virion incorporation of not only HIV-1 envelope glycoprotein but also vesicular stomatitis virus G-glycoprotein through two different pathways. However, the MARCH8 inhibition spectrum remains largely unknown. Here, we show the antiviral spectrum of MARCH8 using viruses pseudotyped with a variety of viral envelope glycoproteins. Infection experiments revealed that viral envelope glycoproteins derived from the rhabdovirus, arenavirus, coronavirus, and togavirus (alphavirus) families were sensitive to MARCH8-mediated inhibition. Lysine mutations at the cytoplasmic tails of rabies virus-G, lymphocytic choriomeningitis virus glycoproteins, SARS-CoV and SARS-CoV-2 spike proteins, and Chikungunya virus and Ross River virus E2 proteins conferred resistance to MARCH8. Immunofluorescence showed impaired downregulation of the mutants of these viral envelope glycoproteins by MARCH8, followed by lysosomal degradation, suggesting that MARCH8-mediated ubiquitination leads to intracellular degradation of these envelopes. Indeed, rabies virus-G and Chikungunya virus E2 proteins proved to be clearly ubiquitinated. We conclude that MARCH8 has inhibitory activity on a variety of viral envelope glycoproteins whose cytoplasmic lysine residues are targeted by this antiviral factor. IMPORTANCE A member of the MARCH E3 ubiquitin ligase family, MARCH8, downregulates many different kinds of host transmembrane proteins, resulting in the regulation of cellular homeostasis. On the other hands, MARCH8 acts as an antiviral factor when it binds to and downregulates HIV-1 envelope glycoprotein and vesicular stomatitis virus G-glycoprotein that are viral transmembrane proteins. This study reveals that, as in the case of cellular membrane proteins, MARCH8 shows broad-spectrum inhibition against various viral envelope glycoproteins by recognizing their cytoplasmic lysine residues, resulting in lysosomal degradation.2022年02月, Microbiology spectrum, 10(1) (1), e0061821, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is a global threat. To forestall the pandemic, developing safe and effective vaccines is necessary. Because of the rapid production and little effect on the host genome, mRNA vaccines are attractive, but they have a relatively low immune response after a single dose. Replicon RNA (repRNA) is a promising vaccine platform for safety and efficacy. RepRNA vaccine encodes not only antigen genes but also the genes necessary for RNA replication. Thus, repRNA is self-replicative and can play the role of an adjuvant by itself, which elicits robust immunity. This study constructed and evaluated a repRNA vaccine in which the gene encoding the spike (S) protein of SARS-CoV-2 was inserted into a replicon of yellow fever virus 17D strain. Upon electroporation of this repRNA into baby hamster kidney cells, the S protein and yellow fever virus protein were co-expressed. Additionally, the self-replication ability of repRNA vaccine was confirmed using qRT-PCR, demonstrating its potency as a vaccine. Immunization of C57BL/6 mice with 1 μg of the repRNA vaccine induced specific T-cell responses but not antibody responses. Notably, the T-cell response induced by the repRNA vaccine was significantly higher than that induced by the nonreplicative RNA vaccine in our experimental model. In the future, it is of the essence to optimize vaccine administration methods and improve S protein expression, like protection of repRNA by nanoparticles and evasion of innate immunity of the host to enhance the immune-inducing ability of the repRNA vaccine.2022年, PloS one, 17(10) (10), e0274829, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), is transmitted mainly by droplet or aerosol infection; however, it may also be transmitted by contact infection. SARS-CoV-2 that adheres to environmental surfaces remains infectious for several days. We herein attempted to inactivate SARS-CoV-2 and influenza A virus adhering to an environmental surface by dry fogging hypochlorous acid solution and hydrogen peroxide solution. SARS-CoV-2 and influenza virus were air-dried on plastic plates and placed into a test chamber for inactivation by the dry fogging of these disinfectants. The results obtained showed that the dry fogging of hypochlorous acid solution and hydrogen peroxide solution inactivated SARS-CoV-2 and influenza A virus in CT value (the product of the disinfectant concentration and contact time)-dependent manners. SARS-CoV-2 was more resistant to the virucidal effects of aerosolized hypochlorous acid solution and hydrogen peroxide solution than influenza A virus; therefore, higher concentrations of disinfectants or longer contact times were required to inactivate SARS-CoV-2 than influenza A virus. The present results provide important information for the development of a strategy that inactivates SARS-CoV-2 and influenza A virus on environmental surfaces by spatial fogging.2022年, PloS one, 17(4) (4), e0261802, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Dengue virus (DENV), from the genus flavivirus of the family flaviviridae, causes serious health problems globally. Human monoclonal antibodies (HuMAb) can be used to elucidate the mechanisms of neutralization and antibody-dependent enhancement (ADE) of DENV infections, leading to the development of a vaccine or therapeutic antibodies. Here, we generated eight HuMAb clones from an Indonesian patient infected with DENV. These HuMAbs exhibited the typical characteristics of weak neutralizing antibodies including high cross-reactivity with other flaviviruses and targeting of the fusion loop epitope (FLE). However, one of the HuMAbs, 3G9, exhibited strong neutralization (NT50 < 0.1 μg/ml) and possessed a high somatic hyper-mutation rate of the variable region, indicating affinity-maturation. Administration of this antibody significantly prolonged the survival of interferon-α/β/γ receptor knockout C57BL/6 mice after a lethal DENV challenge. Additionally, Fc-modified 3G9 that had lost their in vitro ADE activity showed enhanced therapeutic potency in vivo and competed strongly with an ADE-prone antibody in vitro. Taken together, the affinity-matured FLE-targeting antibody 3G9 exhibits promising features for therapeutic application including a low NT50 value, potential for treatment of various kinds of mosquito-borne flavivirus infection, and suppression of ADE. This study demonstrates the therapeutic potency of affinity-matured FLE-targeting antibodies.2021年06月, Scientific reports, 11(1) (1), 12987 - 12987, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- (一社)日本感染症学会, 2021年04月, 感染症学雑誌, 95(臨増) (臨増), 197 - 197, 日本語インドネシアにおける鶏肉由来のキノロン耐性non-typhoidal Salmonella entericaの遺伝子解析
- (一社)日本感染症学会, 2021年04月, 感染症学雑誌, 95(臨増) (臨増), 241 - 241, 日本語インドネシアにおけるヒト及び環境由来のESBL産生Escherichia coliの分布調査
- The analysis of neutralizing epitope of dengue virus (DENV) is important for the development of an effective dengue vaccine. A potent neutralizing mouse monoclonal antibody named 7F4 was previously reported and, here, we further analyzed the detailed epitope of this antibody. 7F4 recognized a novel conformational epitope close to the N-67 glycan on the envelope protein. This antibody was specific to the DENV that lacks N-67 glycan, including the Mochizuki strain. Interestingly, the Mochizuki strain acquired N-67 glycan by 7F4 selective pressure. Considering that most of the currently circulating DENVs possess N-67 glycan, DENVs may have evolved to escape from antibodies targeting 7F4 epitope, suggesting the potency of this neutralizing epitope. In addition, this study demonstrated the existence of the epitopes close to 7F4 epitope and their crucial role in neutralization. In conclusion, the epitopes close to the N-67 glycan are attractive targets for the dengue vaccine antigen. Further analysis of this epitope is warranted.2021年03月, Virus research, 294, 198278 - 198278, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- The development of specific antiviral compounds to SARS-CoV-2 is an urgent task. One of the obstacles for the antiviral development is the requirement of biocontainment because infectious SARS-CoV-2 must be handled in a biosafety level-3 laboratory. Replicon, a non-infectious self-replicative viral RNA, could be a safe and effective tool for antiviral evaluation. Herein, we generated a PCR-based SARS-CoV-2 replicon. Eight fragments covering the entire SARS-CoV-2 genome except S, E, and M genes were amplified with HiBiT-tag sequence by PCR. The amplicons were ligated and in vitro transcribed to RNA. The cells electroporated with the replicon RNA showed more than 3000 times higher luminescence than MOCK control cells at 24 h post-electroporation, indicating robust translation and RNA replication of the replicon. The replication was drastically inhibited by remdesivir, an RNA polymerase inhibitor for SARS-CoV-2. The IC50 of remdesivir in this study was 0.29 μM, generally consistent to the IC50 obtained using infectious SARS-CoV-2 in a previous study (0.77 μM). Taken together, this system could be applied to the safe and effective antiviral evaluation without using infectious SARS-CoV-2. Because this is a PCR-based and transient replicon system, further improvement including the establishment of stable cell line must be achieved.2021年01月, Scientific reports, 11(1) (1), 2229 - 2229, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- JC polyomavirus (JCPyV) causes progressive multifocal leukoencephalopathy (PML), a demyelinating disease of the central nervous system affecting immunocompromised patients. The study of PML-type JCPyV in vitro has been limited owing to the inefficient propagation of the virus in cultured cells. In this study, we carried out long-term culture of COS-7 cells (designated as COS-IMRb cells) transfected with PML-type M1-IMRb, an adapted viral DNA with a rearranged non-coding control region (NCCR). The JCPyV derived from COS-IMRb cells were characterized by analyzing the viral replication, amount of virus by hemagglutination (HA), production of viral protein 1 (VP1), and structure of the NCCR. HA assays indicated the presence of high amounts of PML-type JCPyV in COS-IMRb cells. Immunostaining showed only a small population of JCPyV carrying COS-IMRb cells to be VP1-positive. Sequencing analysis of the NCCR of JCPyV after long-term culture revealed that the NCCR of M1-IMRb was conserved in COS-IMRb cells without any point mutation. The JCPyV genomic DNA derived from a clone of COS-IMRb-3 cells was detected, via Southern blotting, as a single band of approximately 5.1 kbp without deletion. These findings suggest the potential of using COS-IMRb-3 cells as a useful tool for screening anti-JCPyV drugs.2021年01月, Japanese journal of infectious diseases, 74(1) (1), 48 - 53, 英語, 国内誌[査読有り]研究論文（学術雑誌）
- The HIV type 1 (HIV-1) epidemic has continued to grow in Indonesia; however, continuous updates on the epidemiology of HIV-1 in Indonesia remain challenging because it is the biggest archipelago in the world. Furthermore, the emergence of HIV drug resistance (HIVDR) has had a negative impact on the treatment of infected individuals. In this study, we performed HIV-1 subtyping and the detection of HIVDR in 105 HIV-1-infected individuals residing in various cities in Indonesia during 2018-2019. The results obtained identified CRF01_AE as the major epidemic HIV-1 strain, responsible for 81.9% of infection cases, followed by subtype B (12.4%), CRF02_AG (3.8%), CRF52_01B (1%), and a recombinant between CRF01_AE and CRF02_AG (1.0%). Major drug resistance-associated mutations against reverse transcriptase inhibitors were detected in 20% of samples. These results suggest that CRF01_AE is a major HIV-1 strain in Indonesia, while CRF02_AG is emerging. The prevalence of HIVDR in Indonesia needs to be monitored.2020年11月, AIDS research and human retroviruses, 36(11) (11), 957 - 963, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- HIV-1 cure strategy by means of proviral knock-out using CRISPR-Cas9 has been hampered by the emergence of viral resistance against the targeting guide RNA (gRNA). Here, we proposed multiple, concentrated gRNA attacks against HIV-1 regulatory genes to block viral escape. The T cell line were transduced with single and multiple gRNAs targeting HIV-1 tat and rev using lentiviral-based CRISPR-Cas9, followed by replicative HIV-1NL4-3 challenge in vitro. Viral p24 rebound was observed for almost all gRNAs, but multiplexing three tat-targeting gRNAs maintained p24 suppression and cell viability, indicating the inhibition of viral escape. Multiplexed tat gRNAs inhibited acute viral replication in the 2nd round of infection, abolished cell-associated transmission to unprotected T cells, and maintained protection through 45 days, post-infection (dpi) after a higher dose of HIV-1 infection. Finally, we describe here for the first time the assembly of all-in-one lentiviral vectors containing three and six gRNAs targeting tat and rev. A single-vector tat-targeting construct shows non-inferiority to the tat-targeting multi-vector in low-dose HIV-1 infection. We conclude that Cas9-induced, DNA repair-mediated mutations in tat are sufficiently deleterious and deplete HIV-1 fitness, and multiplexed disruption of tat further limits the possibility of an escape mutant arising, thus elevating the potential of CRISPR-Cas9 to achieve a long-term HIV-1 cure.2020年10月, Viruses, 12(11) (11), 英語, 国際誌[査読有り]研究論文（学術雑誌）
- The Dominance of CRF01_AE and the Emergence of Drug Resistance Mutations Among Antiretroviral Therapy-Experienced, HIV-1-infected Individuals in Medan, Indonesia.BACKGROUND: human immunodeficiency virus type 1 (HIV-1) infection is a serious public health threat worldwide. Medan is one example of big cities in Indonesia with a high prevalence of HIV-1 infection; however, quite a limited study had conducted for detecting the circulation of HIV-1 subtypes in Medan. In addition, a serious factor that can implicate the treatment of HIV-1-infected individuals is the emergence of drug resistance mutations. Thus, the information on HIV-1 infection is important to improve the treatment for infected individuals. METHODS: sixty-seven antiretroviral therapy-experienced, HIV-1-infected individuals were recruited for this study. HIV-1 pol genes encoding protease (PR genes) and reverse transcriptase (RT gene), as well as env and gag genes, were amplified from DNA derived from peripheral blood samples. HIV-1 subtyping was conducted to study the dominant HIV-1 subtype circulating in the region. In addition, the emergence of drug resistance mutations was analyzed based on the guidelines published by the International Antiviral Society-United States of America (IAS-USA). RESULTS: the dominant HIV-1 subtype found in Medan was CRF01_AE (77.6%). In addition, another subtype and recombinant viruses such as recombinants between CRF01_AE and subtype B (12.2%), subtype B (4.1%), and CRF02_AG (4.1%) were also found. Drug resistance-associated major mutations were found in 21.6% (8/37) of RT genes and 3.1% (1/32) of PR genes studied. CONCLUSION: our study showed that the dominant subtype found in ART-experienced, HIV-1-infected individuals residing in Medan was CRF01_AE. The emergence of drug resistance mutations in RT and PR genes indicated the importance to monitor the prevalence of drug resistance mutations among HIV-1-infected individuals in Medan.2020年10月, Acta medica Indonesiana, 52(4) (4), 366 - 374, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Introduction: The present study investigated the HIV-1 subtype classification in addition to prevalence of drug resistance mutations (DRMs) in antiretroviral therapy (ART)-experienced and ART-naïve residents of Pontianak, West Kalimantan, Indonesia. Methods: Whole blood samples collected from 30 HIV-1-infected individuals, comprising 19 ART-experienced and 11 ART-naïve individuals, were subjected to RNA and DNA extraction, followed by HIV-1 genes amplification and sequencing analysis. HIV-1 subtyping was classified on viral pol genes encoding reverse transcriptase (RT gene) and protease (PR gene) accompanied by the env and gag genes. DRMs in the RT and PR genes were also analyzed. Results: CRF01_AE was identified as the predominant circulating recombinant form (CRF) of HIV-1 in both ART-experienced and ART-naïve individuals. In addition, CRF02_AG, subtype B, recombinant virus expressing CRF01_AE and subtype B viral genomic fragments, also recombinant virus containing CRF01_AE and CRF02_AG genomic fragments were also identified. Acquired drug resistance (ADR) was identified in 28.5% of ART-experienced individuals, while no transmitted drug resistance was identified in ART-naïve individuals. Conclusions: This study identified CRF01_AE as the most predominant HIV-1 CRF distributing in Pontianak, Indonesia. The prevalence of ADR is considered to be high; thus, further surveillance is needed in this region.2020年09月, Germs, 10(4) (4), 174 - 183, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- The presence of transmitted drug resistance (TDR) in human immunodeficiency virus type 1 (HIV-1) infected individuals naive to antiretroviral therapy, may affect the effectiveness of treatment. Jakarta, the capital city of Indonesia, recorded the highest number of cumulative HIV infection cases in the country. This study aimed to identify on the appearance of TDR, as well as to identify HIV-1 subtypes circulating among treatment-naive individuals in Jakarta. Whole blood samples collected from 43 HIV-1 infected, treatment-naive individuals. Viral subtyping and drug resistance testing were performed on HIV-1 pol genes amplified using nested polymerase chain reaction. CRF01_AE was detected most frequently in Jakarta (73.08%). Drug resistance-related major mutation was not detected in protease fragments of pol gene, but two major mutations, K103N (6.67%) and Y181C (6.67%), were detected in reverse transcriptase fragments of pol gene. Our results suggest that TDR was emerged in Jakarta at a certain extent, thus further surveillance study to monitor the TDR prevalence and circulating HIV-1 subtypes in this region is considered to be necessary.2020年07月, Infectious disease reports, 12(Suppl 1) (Suppl 1), 8740 - 8740, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- To eradicate human immunodeficiency virus type 1 (HIV-1) infection, a comprehensive strategy including preventive vaccine development is needed. Envelope glycoproteins (Env) play a central role in viral infection and are the major targets of humoral immune responses. Therefore, Env is a candidate vaccine antigen, and its characterization is necessary for vaccine development. The characterization of the transmitted/founder (T/F; i.e., recently infected) virus that is responsible for the establishment of infection and induction of primary anti-HIV-1 immune responses is important. We herein established HIV-1 env clones derived from recently infected Indonesian individuals. All env genes were classified into CRF01_AE. The immunological characterization of env clones was performed by neutralization tests using a series of broadly neutralizing antibodies. The present study is the first to immunologically characterize the CRF01_AE T/F virus circulating in Indonesia.2019年10月, AIDS research and human retroviruses, 36(3) (3), 242 - 247, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Human immunodeficiency virus type 1 (HIV-1) is a major causative agent of acquired immune deficiency syndrome. Subtype C (HIV-1C) is the most prevalent HIV-1 subtype worldwide. Although it is highly prevalent in Nepal, genotypic information on Nepalese HIV-1C is limited. We herein investigated the origin and dynamics of HIV-1C in Nepal. Nearly full-length sequencing of Nepalese HIV-1C strains and phylogenetic analyses were performed. The results obtained showed that Nepalese HIV-1C is closely related to the Indian and southern African strains and the introduction of HIV-1C into Nepal was estimated to be in the mid-1980s. These results suggest that multiple HIV-1C strains entered Nepal in the mid-1980s, and this was followed by a marked increase in the number of infection cases for the next decade. These results reflect the current transmission dynamics of HIV-1C strains in Nepal and provide valuable information for HIV monitoring and vaccine development.2019年09月, AIDS research and human retroviruses, 35(9) (9), 870 - 875, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Human immunodeficiency virus type 1 (HIV-1) and acquired immunodeficiency syndrome (AIDS) represent a major public health concern in Indonesia. Although circulating recombinant form (CRF) 01_AE is a predominant subtype in Indonesia, HIV-1 subtype B (HIV-1B) is also widely prevalent. However, the viral genetic evolution, spatial origins, and patterns of transmission of HIV-1B in Indonesia remain unclear. In the present study, we described the evolutionary characteristics and spatial-temporal transmission networks of HIV-1B in Indonesia. To elucidate the epidemiological link between HIV-1B epidemics in Indonesia and those in the remainder of the world, we conducted phylogenetic analyses of HIV-1B strains in Indonesia. Based on the results obtained, at least three epidemic clades [the Indonesia, United States (US), and China clades] of HIV-1B were found to be prevalent in Indonesia. In order to identify the potential source and transmission route of Indonesian HIV-1B strains, we performed Bayesian analyses and constructed Maximum clade credibility trees of each clade. Although some HIV-1B strains in Indonesia were introduced from Thailand, the prevalent HIV-1B strains appeared to have been directly introduced from Europe or America. Indonesian HIV-1B may have spread via the main dispersal of pandemic HIV-1B strains via the US from the Caribbean region rather than being directly introduced from Africa.2019年09月, Scientific reports, 9(1) (1), 13986 - 13986, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Bali, the first province to report a case of HIV in 1987, was placed sixth among Indonesian provinces with the highest cumulative number of HIV cases in 2017. As a popular tourist destination, the spread of genetic variants of HIV through international travel may become a cause for concern in Bali. Tourism is mostly concentrated in south Bali; thus, HIV in less popular regions in north Bali, such as Buleleng Regency, may have viral characteristics different from that in south Bali. Forty-three protease (PR), 40 reverse transcriptase (RT), 27 gag, and 23 env genes were sequenced from 48 samples derived from antiretroviral treatment-experienced individuals. Subtyping revealed CRF01_AE as the dominant circulating recombinant form of HIV-1 in north Bali. Although no major mutation was detected in PR genes, several major mutations were identified in 4 out of the 40 RT genes (10%), indicating the emergence of HIV-1 drug resistance in this region.2019年08月, AIDS research and human retroviruses, 35(8) (8), 769 - 774, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- The Emergence of HIV-1 Transmitted Drug Resistance Mutations Among Antiretroviral Therapy-naive Individuals in Buleleng, Bali, Indonesia.BACKGROUND: the global scale-up of antiretroviral therapy (ART) is the primary factor contributing to the decline in deaths from acquired immune deficiency syndrome (AIDS)-related illnesses. However, the emergence of transmitted drug resistance (TDR) compromises the effects of ART in treatment-naïve individuals, which may hinder treatment success. The present study aimed to identify the presence of TDR among treatment-naive individuals in Buleleng, Bali, which is currently ranked sixth among Indonesian provinces with the highest cumulative human immunodeficiency virus type 1 (HIV-1) infection cases. METHODS: thirty-nine ART-naive individuals in Buleleng Regency General Hospital were enrolled in the present study. Blood samples from participants were subjected to a genotypic analysis. RESULTS: 28 protease (PR) and 30 reverse transcriptase (RT) genes were successfully amplified and sequenced from 37 samples. HIV-1 subtyping revealed CRF01_AE as the dominant circulating recombinant form in the region. No TDR for PR inhibitors was detected; however, TDR for RT inhibitors was identified in five out of 30 samples (16.7%). CONCLUSION: these results indicate the emergence of TDR among ART-naive individuals in Buleleng, Bali. This issue warrants serious consideration because TDR may hamper treatment success and reduce ART efficacy among newly diagnosed individuals. Continuous surveillance with a larger sample size is necessary to monitor TDR among ART-naive individuals.2019年07月, Acta medica Indonesiana, 51(3) (3), 197 - 204, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Comparison of antibody-dependent cellular cytotoxicity activity elicited in recent and chronic hiv-1-infected thai individualsSubsequent to the establishment of human immunodeficiency virus type 1 (HIV-1) infection, host immune responses including humoral and cellular responses against HIV-1 are induced and maintained throughout disease progression. Antibody-dependent cellular cytotoxicity (ADCC) response plays a keyrole in protecting vaccinees from HIV-1 infection in the Thai RV144 vaccine trial. In addition, anti-HIV-1 ADCC response is involved in controlling and slowing disease progression. However, the level of ADCC response in the course of disease progression especially in recent HIV-1 infection is poorly understood. The study compared levels of anti-HIV-1 ADCC activities between serum samples derived from recent and chronic HIV-1-infected individuals in Thailand using peripheral blood mononuclear cells as effector cells and HIV-1-infected CEM.NKR CCR5+ cells as target cells. Distribution of ADCC activities do not differ significantly between both groups. Anti-HIV-1 ADCC activity was induced in a limited number of recent and chronic HIV-1-infected individuals. However, induction ratio of ADCC activity is not significantly related to the course of disease progression. These results provide additional information on ADCC responses during the course of HIV-1 infection, recent and chronic.SEAMEO TROPMED Network, 2019年07月, Southeast Asian Journal of Tropical Medicine and Public Health, 50(4) (4), 651 - 662, 英語研究論文（学術雑誌）
- Elsevier {BV}, 2019年07月, Journal of Virus Eradication, 5, 17 - 18, 英語, 国際誌, 国際共著している[査読有り]研究論文（学術雑誌）
- Manado, the capital city of North Sulawesi, is a unique region in Indonesia because of its religion. We collected peripheral blood samples from 63 individuals on antiretroviral therapy. The amplification of viral genomic fragments, viral subtyping, detection of HIV drug resistance-associated mutations (DRAMs), and phylogenetic analyses were performed. Viral subtyping revealed that the most prevalent HIV type 1 (HIV-1) subtype/circulating recombinant form (CRF) was CRF01_AE (84.1%), followed by subtype B (6.8%) and recombinants between CRF01_AE and CRF02_AG (4.5%). Although no major DRAMs were present in protease genes, they were detected in reverse transcriptase (RT) genes. Nine of 38 samples (23.7%) had major DRAMs against nucleoside RT inhibitors (NRTIs) and/or non-NRTIs. The results of phylogenetic analyses indicated that CRF01_AE in North Sulawesi is related to that in Bali. Therefore, Bali may play an important role in circulating CRF01_AE in North Sulawesi.2019年04月, AIDS research and human retroviruses, 35(4) (4), 407 - 413, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Two new antibiotics, designated virantmycin B (1) and C (2), were isolated from the cultured broth of Streptomyces sp. AM-2504. Compounds 1 and 2 were purified by Diaion HP-20, silica gel, and octadecylsilane chromatography, followed by high-performance liquid chromatography. The chemical structures of the new compounds, 1 and 2, were determined by nuclear magnetic resonance and mass spectrometry, as containing a terahydroquinoline and an indoline, respectively, each also containing a hydroxy cyclopentenone moiety. Both compounds demonstrated weak antimicrobial (both antibacterial and antifungal) activity and compound 1 also showed antiviral activity against the dengue virus, whereas compound 2 exhibited no antiviral properties.2019年03月, The Journal of antibiotics, 72(3) (3), 169 - 173, 英語, 国内誌[査読有り]研究論文（学術雑誌）
- Antiviral Activity of Cananga odorata Against Hepatitis BChronic hepatitis B virus (HBV) infection can lead to liver cirrhosis and hepatocellular carcinoma. Current therapeutic drugs for chronic hepatitis B using pegylated interferons and nucleos(t)ide analogs have limited efficacy. Therefore, the development of novel and safe antivirals is required. Natural products including medicinal plants produce complex and structurally diverse compounds, some of which offer suitable targets for antiviral screening studies. In the present study, we screened various crude extracts from Indonesian plants for anti-HBV activity by determining their effects on the production of extracellular HBV DNA in Hep38.7-Tet cells and HBV entry onto a HBV-susceptible cell line, HepG2-NTCP, with the following results: (1) In Hep38.7-Tet cells, Cananga odorata exhibited the highest anti-HBV activity with a 50% inhibitory concentration (IC50) of 56.5 µg/ml and 50% cytotoxic concentration (CC50) of 540.2 µg/ml (Selectivity Index: 9.6). (2) The treatment of HepG2-NTCP cells with Cassia fistula, C. odorata, and Melastoma malabathricum at concentrations of 100 µg/ml lowered the levels of HBsAg production to 51.2%, 58.0%, and 40.1%, respectively, compared to untreated controls, and IC50 and CC50 values of C. odorata were 142.9 µg/ml and >400 µg/ml. In conclusion, the C. odorata extract could be a good candidate for the development of anti-HBV drugs.2019年, Kobe Journal of Medical Sciences, 65(2) (2), E71 - E79, 英語, 国内誌[査読有り]研究論文（学術雑誌）
- Human immunodeficiency virus (HIV) infection and acquired immune deficiency syndrome (AIDS) (HIV/AIDS) remain a serious health problem in Indonesia. Jakarta has the largest number of individuals infected with HIV type 1 (HIV-1) in Indonesia; however, data on viral subtyping of HIV-1 is limited. We conducted a molecular epidemiological study on HIV-1 genes derived from infected, drug-naive individuals residing in Jakarta, the capital of Indonesia with large communities of HIV-infected individuals. To determine the distribution of HIV-1 subtypes circulating in Jakarta, 43 HIV-1-infected individuals were recruited in Jakarta, Indonesia. HIV-1 gag and envgenes were amplified from peripheral blood samples derived from study participants. Viral subtyping using the Recombinant Identification Program (RIP) and phylogenetic tree analysis were conducted. RIP and phylogenetic tree analysis revealed that CRF01-AE was the dominant subtype in Jakarta (40%), followed by recombinant viruses containing CRF01-AE and subtype B gene fragments (7%) and subtype B (7%). This study reported the emergence of CRF 01-AE as predominant subtype among HIV-1-infected, drug-naïve individuals in Jakarta, Indonesia. Therefore, continuous surveillance is required in order to detect the emergence of HIV-1-infection.2018年12月, IOP Conference Series: Materials Science and Engineering, 434(1) (1)[査読有り]研究論文（国際会議プロシーディングス）
- The CRISPR/Cas9 system provides a novel and promising tool for editing the HIV-1 proviral genome. We designed RNA-guided CRISPR/Cas9 targeting the HIV-1 regulatory genes tat and rev with guide RNAs (gRNA) selected from each gene based on CRISPR specificity and sequence conservation across six major HIV-1 subtypes. Each gRNA was cloned into lentiCRISPRv2 before co-transfection to create a lentiviral vector and transduction into target cells. CRISPR/Cas9 transduction into 293 T and HeLa cells stably expressing Tat and Rev proteins successfully abolished the expression of each protein relative to that in non-transduced and gRNA-absent vector-transduced cells. Tat functional assays showed significantly reduced HIV-1 promoter-driven luciferase expression after tat-CRISPR transduction, while Rev functional assays revealed abolished gp120 expression after rev-CRISPR transduction. The target gene was mutated at the Cas9 cleavage site with high frequency and various indel mutations. Conversely, no mutations were detected at off-target sites and Cas9 expression had no effect on cell viability. CRISPR/Cas9 was further tested in persistently and latently HIV-1-infected T-cell lines, in which p24 levels were significantly suppressed even after cytokine reactivation, and multiplexing all six gRNAs further increased efficiency. Thus, the CRISPR/Cas9 system targeting HIV-1 regulatory genes may serve as a favorable means to achieve functional cures.Nature Publishing Group, 2018年12月, Scientific Reports, 8(1) (1), 7784, 英語[査読有り]研究論文（学術雑誌）
- JC polyomavirus (JCPyV) is the causative agent of progressive multifocal leukoencephalopathy (PML), a demyelinating disease of the central nervous system in immunocompromised patients. Archetype JCPyV circulates in the human population. There have been several reports of archetype JCPyV replication in cultured cells, in which propagation was not enough to produce high titers of archetype JCPyV. In this study, we carried out cultivation of the transfected cells with archetype JCPyV DNA MY for more than 2 months to establish COS-7 cells (designated COS-JC cells) persistently producing archetype JCPyV. Moreover, JCPyV derived from COS-JC cells was characterized by analyzing the viral propagation, size of the viral genome, amount of viral DNA, production of viral protein, and structure of the non-coding control region (NCCR). Southern blotting using a digoxigenin-labeled JCPyV probe showed two different sizes of the JCPyV genome in COS-JC cells. For molecular cloning, four of five clones showed a decrease in the size of complete JCPyV genome. Especially, clone No. 10 was generated the large deletion within the Large T antigen. On the other hand, the archetype structure of the NCCR was maintained in COS-JC cells, although a few point mutations occurred. Quantitative PCR analysis of viral DNA in COS-JC cells indicated that a high copy number of archetype JCPyV DNA was replicated in COS-JC cells. These findings suggest that COS-JC cells could efficiently propagate archetype JCPyV MY and offer a useful tool to study persistent infection of archetype JCPyV in a kidney-derived system.2018年08月, Microbiology and immunology, 62(8) (8), 524 - 530, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Kepulauan Riau is a famous tourist destination in Indonesia. The epidemic of human immunodeficiency virus type 1 (HIV-1) infection is gradually increasing in this region. We collected peripheral blood samples from 62 antiretroviral therapy-experienced individuals. The amplification of viral genomic fragments, HIV-1 subtyping, and the detection of HIV drug resistance (HIVDR) were performed. Viral subtyping revealed that the most prevalent HIV-1 subtype/circulating recombinant form (CRF) was CRF01-AE (55.6%), followed by recombinants between CRF01-AE and subtype B (17.8%) and then subtype B (15.6%). Recombinants containing CRF02-AG gene fragments were also detected (11.1%). Regarding HIVDR, no drug resistance-Associated major mutations were found in pol genes encoding protease, although minor mutations were frequently detected. Furthermore, major mutations, including M184V (2.2%) and Y188L (2.2%), were identified in the viral pol gene encoding reverse transcriptase derived from a study participant, suggesting that the prevalence of HIVDR is low in the region.Mary Ann Liebert Inc., 2018年06月, AIDS Research and Human Retroviruses, 34(6) (6), 555 - 560, 英語[査読有り]研究論文（学術雑誌）
- 2018年06月, Journal of medicinal chemistry, 61(12) (12), 5138 - 5153[査読有り]
- Introduction The April 2015 Nepal earthquake resulted in more than 8,700 deaths and 22,000 casualties including damage to health facilities. The impact of this situation on chronic conditions such as human immunodeficiency virus infection and acquired immune deficiency syndrome (HIV/AIDS) may become a long-lasting public health threat. Therefore, the objectives of this study were i) to assess the association of antiretroviral therapy (ART) adherence with mental health problems, and social behaviors, ii) to examine factors affecting treatment failure, and iii) to investigate changes in ART adherence and post-traumatic stress disorder (PTSD) among people living with HIV 6 and 12 months after the disaster. Methods Study was conducted 6 months after the earthquake in 2015 with a sample size of 305 earthquake victims with HIV and followed after 12 months of the earthquake. A logistic regression analysis was used to examine relationships, while a paired t-test analysis was conducted to assess changes in adherence to ART and PTSD level at 6 months and 12 months after earthquake. Results In the earthquake, 5.2% of the participants lost their family member. Approximately 44% of participants had earthquake-PTSD symptoms and 50% experienced HIV stigma. PTSD and HIV status disclosure were significantly associated with adherence to ART, while HIV stigma and religion were associated with treatment failure. PTSD and adherence levels to ART were significantly improved over the 6-month period. Conclusion Awareness programs for general public to eliminate HIV stigma promote psychosocial counseling to earthquake victims living with HIV in order to reduce PTSD will contribute to maintaining optimal ART adherence and to prevent treatment failure.Public Library of Science, 2018年06月, PLoS ONE, 13(6) (6), e0198071, 英語[査読有り]研究論文（学術雑誌）
- 2018年05月, HIV AIDS Rev, 17(2) (2), 81 - 90, 英語Genotypic characterization of human immunodeficiency virus type 1 isolated in Bali, Indonesia in 2016.[査読有り]研究論文（学術雑誌）
- Aedes aegypti and Aedes albopictus are the primary and secondary vectors, respectively, of dengue, the most important arboviral disease in the world. The aim of this study was to detect and serotype dengue viruses (DENV) in the vectors Ae. aegypti and Ae. albopictus in Surabaya, Indonesia. Between 2008 and 2015, 16,605 Aedes mosquitoes were collected in 15 sub-districts of Surabaya. Ae. aegypti was dominant (90.9%), whereas few Ae. albopictus were collected (9.1%). A total of 330 pools of adult Aedes mosquitoes were subjected to the serotyping of DENV by RT-PCR. DENV-1 (52.3%) was the most frequently detected serotype, followed by DENV-2 (40.3%), DENV-4 (4.6%), and DENV-3 (2.8%). The average minimum infection rate for Ae. aegypti in various sub-districts of Surabaya was 7.2 per 1,000 mosquitoes, while that for Ae. albopictus was 0.7 per 1,000 mosquitoes. The results showed that the predominantly circulating DENV serotype in mosquitoes continuously shifted from DENV-2 (2008) to DENV-1 (2009-2012), to DENV-2 again (2013-2014), and then back to DENV-1 (2015). The circulating DENV serotypes in mosquitoes were generally consistent with those in humans. Therefore, the surveillance of infected mosquitoes with DENV might provide an early warning sign for the risk of future dengue outbreaks.National Institute of Health, 2018年, Japanese Journal of Infectious Diseases, 71(1) (1), 58 - 61, 英語[査読有り]研究論文（学術雑誌）
- The global occurrence of viral infectious diseases poses a significant threat to human health. Dengue virus (DENV) infection is one of the most noteworthy of these infections. According to a WHO survey, approximately 400 million people are infected annually symptoms deteriorate in approximately one percent of cases. Numerous foundational and clinical investigations on viral epidemiology, structure and function analysis, infection source and route, therapeutic targets, vaccines, and therapeutic drugs have been conducted by both academic and industrial researchers. At present, CYD-TDV or Dengvaxia® is the only approved vaccine, but potent inhibitors are currently under development. In this review, an overview of the viral life circle and the history of DENVs is presented, and the most recently reported antiviral candidates and newly discovered promising targets are focused and summarized. We believe that these successes and failures have enabled progress in anti-DENV drug discovery and hope that our review will stimulate further innovation in this area.Pharmaceutical Society of Japan, 2018年, Chemical and Pharmaceutical Bulletin, 66(3) (3), 191 - 206, 英語[査読有り]
- Background: Dengue is a kind of infectious disease that was distributed in the tropical and sub-tropical areas. To date, there is no clinically approved dengue vaccine or antiviral for humans, even though there have been great efforts towards this end. Therefore, finding the effective compound against dengue virus (DENV) replication is very important. Among the complex compounds, copper(II)-imidazole derivatives are of interest because of their biological and medicinal benefits. Materials and Methods: In the present study, antiviral activity of [Cu(2,4,5-triphenylimidazole)2]n, was evaluated against different stages of dengue virus type 2 (DENV-2) replication in Vero cell using focus forming unit reduction assay and quantitative ELISA. Results: [Cu(2,4,5-triphenylimidazole)2]n inhibited DENV-2 replication in Vero cells with IC50 = 2.3 μg/ml and SI= 19.42 when cells were treated 2 days after virus infection, whereas its CC50 for cytotoxicity to Vero cells was 44.174 μg/ml. Conclusion: The compound has high anti-DENV2 activity, less toxicity, and a high possibility to be considered a drug candidate.Obafemi Awolowo University, 2018年, African Journal of Infectious Diseases, 12(1) (1), 116 - 119, 英語[査読有り]研究論文（学術雑誌）
- Dengue virus (DENV) infection is a major health issue in tropical and subtropical areas. Indonesia is one of the biggest dengue endemic countries in the world. In the present study, the phylogenetic analysis of DENV in Bangkalan, Madura Island, Indonesia, was performed in order to obtain a clearer understanding of its dynamics in this country. A total of 359 blood samples from dengue-suspected patients were collected between 2012 and 2014. Serotyping was conducted using a multiplex Reverse Transcriptase-Polymerase Chain Reaction and a phylogenetic analysis of E gene sequences was performed using the Bayesian Markov chain Monte Carlo (MCMC) method. 17 out of 359 blood samples (4.7%) were positive for the isolation of DENV. Serotyping and the phylogenetic analysis revealed the predominance of DENV-1 genotype I (9/17, 52.9%), followed by DENV-2 Cosmopolitan type (7/17, 41.2%) and DENV-3 genotype I (1/17, 5.9%). DENV-4 was not isolated. The Madura Island isolates showed high nucleotide similarity to other Indonesian isolates, indicating frequent virus circulation in Indonesia. The results of the present study highlight the importance of continuous viral surveillance in dengue endemic areas in order to obtain a clearer understanding of the dynamics of DENV in Indonesia.2018年, Journal of tropical medicine, 2018, 8127093 - 8127093, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- BACKGROUND AND OBJECTIVES: Human Immunodeficiency Virus (HIV) is still a major health issue in Indonesia. In recent years, the appearance of drug resistance-associated mutations has reduced the effectiveness of Antiretroviral Therapy (ART). We conducted genotypic studies, including the detection of drug resistance-associated mutations (from first-line regimen drugs), on HIV-1 genes derived from infected individuals in Maumere, West Nusa Tenggara. Maumere, a transit city in West Nusa Tenggara, which has a high HIV-1 transmission rate. METHOD: We collected 60 peripheral blood samples from 53 ART-experienced and 7 ART-naive individuals at TC Hillers Hospital, Maumere between 2014 and 2015. The amplification and a sequencing analysis of pol genes encoding protease (the PR gene) and reverse transcriptase (the RT gene) as well as the viral env and gag genes were performed. HIV-1 subtyping and the detection of drug resistance-associated mutations were then conducted. RESULTS: Among 60 samples, 46 PR, 31 RT, 30 env, and 20 gag genes were successfully sequenced. The dominant HIV-1 subtype circulating in Maumere was CRF01_AE. Subtype B and recombinant viruses containing gene fragments of CRF01_AE, subtypes A, B, C, and/or G were also identified as minor populations. The major drug resistance-associated mutations, M184V, K103N, Y188L, and M230I, were found in the RT genes. However, no major drug resistance-associated mutations were detected in the PR genes. CONCLUSION: CRF01_AE was the major HIV-1 subtype prevalent in Maumere. The appearance of drug resistance-associated mutations found in the present study supports the necessity of monitoring the effectiveness of ART in Maumere.2018年, Current HIV research, 16(2) (2), 158 - 166, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- The envelope glycoprotein (Env) gp120 of human immunodeficiency virus type 1 (HIV-1) plays a critical role in viral entry into host cells. The broadly neutralizing human monoclonal antibody VRC01, which recognizes the CD4 binding site on gp120, neutralizes more than 90% of HIV-1 isolates. However, some of the CRF01_AE viruses prevalent in Southeast Asia are resistant to VRC01-mediated neutralization. We previously reported that 3 amino acid residues at positions 185, 186, and 197 of gp120 played an important role in the VRC01 resistance of CRF01_AE Env (AE-Env) clones isolated from HIV-infected Thai individuals. However, the VRC01 susceptibility of AE-Env clones was not fully explained by mutations at these 3 residues. In the present study, we examined other factors involved in the acquisition of viral VRC01 resistance. Neutralization tests using lentiviral vectors expressing a series of mutant AE-Env clones revealed that the deletion of 2-4 amino acid residues on the loop structure in the V5 region of gp120 conferred VRC01 resistance to several AE-Env clones. Our results provide novel insights into the mechanisms underlying viral VRC01 resistance.MARY ANN LIEBERT, INC, 2017年12月, AIDS RESEARCH AND HUMAN RETROVIRUSES, 33(12) (12), 1248 - 1257, 英語[査読有り]研究論文（学術雑誌）
- Hepatitis C virus (HCV), dengue virus (DENV) and Japanese encephalitis virus (JEV) belong to the family Flaviviridae. Their viral particles have the envelope composed of viral proteins and a lipid bilayer acquired from budding through the endoplasmic reticulum (ER). The phospholipid content of the ER membrane differs from that of the plasma membrane (PM). The phospholipase A(2) (PLA(2)) superfamily consists of a large number of members that specifically catalyse the hydrolysis of phospholipids at a particular position. Here we show that the CM-II isoform of secreted PLA(2) obtained from Naja mossambica mossambica snake venom (CM-II-sPLA(2)) possesses potent virucidal (neutralising) activity against HCV, DENV and JEV, with 50% inhibitory concentrations (IC50) of 0.036, 0.31 and 1.34 ng/ ml, respectively. In contrast, the IC50 values of CM-II-sPLA(2) against viruses that bud through the PM (Sindbis virus, influenza virus and Sendai virus) or trans-Golgi network (TGN) (herpes simplex virus) were > 10,000 ng/ml. Moreover, the 50% cytotoxic (CC50) and haemolytic (HC50) concentrations of CMII- sPLA(2) were > 10,000 ng/ml, implying that CM-II-sPLA(2) did not significantly damage the PM. These results suggest that CM-II-sPLA(2) and its derivatives are good candidates for the development of broadspectrum antiviral drugs that target viral envelope lipid bilayers derived from the ER membrane.NATURE PUBLISHING GROUP, 2017年11月, SCIENTIFIC REPORTS, 7(1) (1), 15931, 英語[査読有り]研究論文（学術雑誌）
- Molecular epidemiological data on human immunodeficiency virus type 1 (HIV-1) are limited in Nepal and have not been available in areas affected by the April 2015 earthquake. Therefore, we conducted a genotypic study on HIV-1 genes derived from individuals on antiretroviral therapy residing in 14 districts in Nepal highly affected by the earthquake. HIV-1 genomic fragments were amplified from 40 blood samples of HIV treatment-failure individuals, and a sequencing analysis was performed on these genes. In the 40 samples, 29 protease, 32 reverse transcriptase, 25 gag, and 21 env genes were sequenced. HIV-1 subtyping revealed that subtype C (84.2%, 32/38) was the major subtype prevalent in the region, while CRF01_AE (7.9%, 3/38) and other recombinant forms (7.9%, 3/38) were also detected. In addition, major drug resistance mutations were identified in 21.9% (7/32) of samples, indicating the possible emergence of HIV-1 drug resistance in earthquake-affected areas in Nepal.MARY ANN LIEBERT, INC, 2017年09月, AIDS RESEARCH AND HUMAN RETROVIRUSES, 33(9) (9), 960 - 965, 英語[査読有り]研究論文（学術雑誌）
- 2017年07月, Acta medica Indonesiana, 49(3) (3), 205 - 214Sero- and Molecular Epidemiology of HIV-1 in Papua Province, Indonesia.[査読有り]研究論文（学術雑誌）
- JC polyomavirus (JCPyV) is the causative agent of the demyelinating disease of the central nervous system known as progressive multifocal leukoencephalopathy (PML), which occurs in immunocompromised patients. Moreover, patients treated with natalizumab for multiple sclerosis or Crohn disease can develop PML, which is then termed natalizumab-related PML. Because few drugs are currently available for treating PML, many antiviral agents are being investigated. It has been demonstrated that the topoisomerase I inhibitors topotecan and -lapachone have inhibitory effects on JCPyV replication in IMR-32 cells. However, both of these drugs have marginal inhibitory effects on virus propagation in JC1 cells according to RT-PCR analysis. In the present study, the inhibitory effect of another topoisomerase I inhibitor, 7-ethy-10-[4-(1-piperidino)-1-piperidino] carbonyloxy camptothecin (CPT11), was assessed by investigating viral replication, propagation, and viral protein 1 (VP1) production in cultured cells. JCPyV replication was assayed using real-time PCR combined with Dpn I treatment in IMR-32 cells transfected with JCPyV DNA. It was found that JCPyV replicates less in IMR-32 cells treated with CPT11 than in untreated cells. Moreover, CPT11 treatment of JCI cells persistently infected with JCPyV led to a dose-dependent reduction in JCPyV DNA and VP1 production. Additionally, the inhibitory effect of CPT11 was found to be stronger than those of topotecan and -lapachone. These findings suggest that CPT11 may be a potential anti-JCPyV agent that could be used to treat PML.WILEY, 2017年06月, MICROBIOLOGY AND IMMUNOLOGY, 61(6) (6), 232 - 238, 英語[査読有り]研究論文（学術雑誌）
- JC polyomavirus (JCPyV) is the causative agent of the demyelinating disease of the central nervous system known as progressive multifocal leukoencephalopathy (PML), which occurs in immunocompromised patients. Moreover, patients treated with natalizumab for multiple sclerosis or Crohn disease can develop PML, which is then termed natalizumab-related PML. Because few drugs are currently available for treating PML, many antiviral agents are being investigated. It has been demonstrated that the topoisomerase I inhibitors topotecan and β-lapachone have inhibitory effects on JCPyV replication in IMR-32 cells. However, both of these drugs have marginal inhibitory effects on virus propagation in JC1 cells according to RT-PCR analysis. In the present study, the inhibitory effect of another topoisomerase I inhibitor, 7-ethy-10-[4-(1-piperidino)-1-piperidino] carbonyloxy camptothecin (CPT11), was assessed by investigating viral replication, propagation, and viral protein 1 (VP1) production in cultured cells. JCPyV replication was assayed using real-time PCR combined with Dpn I treatment in IMR-32 cells transfected with JCPyV DNA. It was found that JCPyV replicates less in IMR-32 cells treated with CPT11 than in untreated cells. Moreover, CPT11 treatment of JCI cells persistently infected with JCPyV led to a dose-dependent reduction in JCPyV DNA and VP1 production. Additionally, the inhibitory effect of CPT11 was found to be stronger than those of topotecan and β-lapachone. These findings suggest that CPT11 may be a potential anti-JCPyV agent that could be used to treat PML.2017年06月, Microbiology and immunology, 61(6) (6), 232 - 238, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- HIV disease became a manageable chronic disease since combination antiretroviral therapy (cART) was introduced as the standard treatment regimen. However, the emergence of drug-resistant viruses is a major problem associated with cART. A phenotypic drug susceptibility test using a lentiviral vector was established and applied to evaluate new protease inhibitors (PIs). Lentiviral vectors representing a wild-type (WT-lentivector) and darunavir (DRV)-resistant HIV type 1 (HIV-1) (DRVr -lentivector) were generated. Nine clinically approved protease inhibitors (PIs) inhibited the transduction ability of WT-lentivector similar to their inhibitory effects on the replication of WT HIV-1. Three new PIs reduced the transduction ability of WT- and DRVr -lentivector, suggesting that these PIs may be the candidates as novel antiretroviral drugs against drug-resistant variants of HIV-1.WILEY, 2017年01月, FEBS OPEN BIO, 7(1) (1), 88 - 95, 英語[査読有り]研究論文（学術雑誌）
- The high impact of the acquired immunodeficiency syndrome caused by human immunodeficiency virus has represented a significant public health threat in the last four decades. Recently, the development of new antiviral drugsiMedPub Journals, 2016年11月, Journal of Prevention & Infection Control, 2(2:12) (2:12), 1 - 5, 英語
has led to great progress in overcoming this virus. Antiretroviral therapy coverage has been increasing, even in low-income countries, resulting in an overall decline of HIV/AIDS-related deaths worldwide. Nonetheless, new
infections still appear, and the era of clearance of this virus has not yet been reached. Moreover, the resistance of the virus to drugs during treatments remains a serious problem.
Therefore, researchers are actively seeking to discover and characterize new molecule(s) that could be suitable for anti-HIV treatment. Cyclophilin A, a host molecular chaperone or folder protein, is suspected to be one such attractive target. We and others are searching for inhibitor(s) of this molecule and investigating its mechanism of action throughout the viral life cycle. In this review, we focus on studies related to cyclophilin A and the candidate inhibitors.[査読有り] - NATL INST INFECTIOUS DISEASES, 2016年09月, JAPANESE JOURNAL OF INFECTIOUS DISEASES, 69(5) (5), 442 - 444, 英語[査読有り]研究論文（学術雑誌）
- Papua and West Papua provinces have the highest prevalence rate of human immunodeficiency virus type 1 (HIV-1) infection in Indonesia; however, data on the molecular epidemiology of HIV-1 are limited. We conducted a genotypic study on HIV-1 genes derived from antiretroviral therapy-naive individuals residing in Sorong, West Papua. HIV-1 genomic fragments were amplified from 43 peripheral blood samples, and sequencing analysis of the genes was carried out. Of the 43 samples, 41 protease (PR), 31 reverse transcriptase (RT), 26 gag, and 25 env genes were sequenced. HIV-1 subtyping revealed that CRF01_AE (48.8%, 21/43) and subtype B (41.9%, 18/43) were the major subtypes prevalent in the region, whereas other recombinant forms were also detected. Major drug resistance-associated mutations for PR inhibitors were not detected; however, mutations for the RT inhibitors, A62V and E138A, appeared in a few samples, indicating the possible emergence of transmitted HIV-1 drug resistance in Sorong, West Papua.MARY ANN LIEBERT, INC, 2016年08月, AIDS RESEARCH AND HUMAN RETROVIRUSES, 32(8) (8), 812 - 817, 英語[査読有り]研究論文（学術雑誌）
- Neutralizing antibody responses play important roles in controlling several viral infections including human immunodeficiency virus type 1 (HIV-1). Potent and broad neutralizing antibody responses have been reported in some HIV-1-infected individuals; therefore, elucidating the mechanisms underlying neutralizing antibody responses will provide important information for the development of anti-HIV-1 vaccines. We herein performed a comparative study on the neutralization breadth and potency of serum samples collected from Thai individuals recently and chronically infected with HIV-1. Neutralization tests using a series of envelope glycoproteins (Env)-recombinant viruses revealed that although several serum samples derived from recently infected individuals did not show any HIV-1-specific neutralizing activity, the remaining serum samples exhibited neutralizing activity not only for recombinant viruses with CRF01_AE Env, but also for viruses with subtypes B and C Env. Furthermore, some serum samples derived from recently infected individuals showed the neutralization potency. Our results may provide a deeper insight into the characteristics of neutralizing antibody responses that develop during the course of HIV-1 infection among individuals in Thailand. (C) 2016 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.ELSEVIER SCIENCE BV, 2016年05月, MICROBES AND INFECTION, 18(5) (5), 346 - 353, 英語[査読有り]研究論文（学術雑誌）
- Dengue virus (DENV) infection-enhancing antibodies are a hypothetic factor to increase the dengue disease severity. In this study, we investigated the enhancing antibodies against Indonesian strains of DENY-1-4 in 50 healthy inhabitants of central Thailand (Bangkok and Uthai Thani). Indonesia and Thailand have seen the highest dengue incidence in Southeast Asia. The infection history of each subject was estimated by comparing his/her neutralizing antibody titers against prototype DENV-1-4 strains. To resolve the difficulty in obtaining foreign live viruses for use as assay antigens, we used a recombinant system to prepare single-round infectious dengue viral particles based on viral sequence information. Irrespective of the previously infecting serotype(s), most serum samples showed significantly higher enhancement titers against Indonesian DENV-2 strains than against Thai DENV-2 strains, whereas the opposite effect was observed for the DENV-3 strains. Equivalent enhancing activities were observed against both DENV-1 and DENV-4. These results suggest that the genotype has an impact on enhancing antibody activities against DENV-2 and DENV-3, because the predominant circulating genotypes of each serotype differ between Indonesia and Thailand. (C) 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.ELSEVIER SCIENCE BV, 2016年04月, MICROBES AND INFECTION, 18(4) (4), 277 - 284, 英語[査読有り]研究論文（学術雑誌）
- JC polyomavirus (JCPyV) causes progressive multifocal leukoencephalopathy (PML), a fatal demyelinating disease of the central nervous system, in immunocompromised patients. Because no drugs have been approved for treating PML, many antiviral agents are currently being investigated for this purpose. The inhibitory effects of the topoisomerase I inhibitors topotecan and -lapachone were assessed by investigating viral replication, propagation and viral protein 1 (VP1) production in cultured cells. JCPyV replication was assayed using the human neuroblastoma cell line IMR-32 transfected with the JCPyV plasmid and RT- PCR combined with Dpn I treatment. Dpn I digests the input plasmid DNA containing methylated adenosine, but not newly replicated JCPyV DNA, in IMR-32 cells. It was found that JCPyV replicates less in IMR-32 cells treated with topotecan or -lapachone than in untreated cells. Moreover, drug treatment of JCI cells, which are IMR-32 cells persistently infected with JCPyV, led to a reduction in the amount of JCPyV DNA and population of VP1-positive cells. These results demonstrate that topotecan and -lapachone affects JCPyV propagation in human neuroblastoma cell lines, suggesting that topotecan and -lapachone could potentially be used to treat PML.WILEY-BLACKWELL, 2016年04月, MICROBIOLOGY AND IMMUNOLOGY, 60(4) (4), 253 - 260, 英語[査読有り]研究論文（学術雑誌）
- Indonesia is one of the biggest dengue endemic countries, and, thus, is an important place to investigate the evolution of dengue virus (DENV). We have continuously isolated DENV in Surabaya, the second biggest city in Indonesia, since 2008. We previously reported sequential changes in the predominant serotype from DENV type 2 (DENV-2) to DENV type 1 (DENV-1) in November 2008 and from DENV-1 to DENV-2 in July 2013. The predominance of DENV-2 continued in 2014, but not in 2015. We herein phylogenetically investigated DENV-2 transitions in Surabaya between 2008 and 2014 to analyze the divergence and evolution of DENV-2 concomitant with serotype shifts. All DENV-2 isolated in Surabaya were classified into the Cosmopolitan genotype, and further divided into 6 clusters. Clusters 1-3, dominated by Surabaya strains, were defined as the "Surabaya lineage". Clusters 4-6, dominated by strains from Singapore, Malaysia, and many parts of Indonesia, were the "South East Asian lineage". The most recent common ancestor of these strains existed in 1988, coinciding with the time that an Indonesian dengue outbreak took place. Cluster 1 appeared to be unique because no other DENV-2 isolate was included in this cluster. The predominance of DENV-2 in 2008 and 2013-14 were caused by cluster 1, whereas clusters 2 and 3 sporadically emerged in 2011 and 2012. The characteristic amino acids of cluster 1, E-170 V and E-282Y, may be responsible for its prevalence in Surabaya No amino acid difference was observed in the envelope region between strains in 2008 and 2013-14, suggesting that the re-emergence of DENV-2 in Surabaya was due to the loss or decrease of herd immunity in the 5-year period when DENV-2 subsided. The South East Asian lineage primarily emerged in Surabaya in 2014, probably imported from other parts of Indonesia or foreign countries. (C) 2015 Elsevier B.V. All rights reserved.ELSEVIER SCIENCE BV, 2016年01月, INFECTION GENETICS AND EVOLUTION, 37, 88 - 93, 英語[査読有り]研究論文（学術雑誌）
- The development of gp120 targeted human immunodeficiency virus (HIV) drug has improved antiretroviral therapies owing to its effects on attachment to target cells. Some currently available antiretroviral therapies that act by inhibiting viral infection still are limited on the toxicity issues; therefore, novel approaches to treat and prevent HIV infections are still needed. Herein, we introduce carbon dot nanoparticles as a new strategy for preventing HIV-1 infection via interaction with gp120 and subsequent elimination of target cell interaction. Carbon dots (diameter: similar to 2 nm) exhibiting a graphene-like structure were prepared by pyrolysis of citric acid and further associated with boronic acid-containing molecules. Specific peaks from Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy analysis indicated successful modification of the carbon dots by boronic acid. The lower cytotoxic effects of this carbon-based material were evaluated using WST-1 assays. The existence of boronic acid moieties on the edge of carbon dots enhanced the inhibitory activity by suppressing syncytium formation. These findings provide a basis for further studies of carbon dot-based applications in HIV prevention and therapy.ROYAL SOC CHEMISTRY, 2016年, RSC ADVANCES, 6(95) (95), 92996 - 93002, 英語[査読有り]研究論文（学術雑誌）
- Universitas Airlangga, 2015年07月, Indonesian Journal of Tropical and Infectious Disease, 5(1) (1), 1 - 1研究論文（学術雑誌）
- {OMICS} Publishing Group, 2015年05月, J. Infect. Dis. Ther., 3(3) (3), 1 - 8, 英語[査読有り]研究論文（学術雑誌）
- It has been difficult to study JCV replication because of its restricted host range. In this study, JCV replication was examined using different clones in 293cells. RT-PCR assay revealed that large T antigen expression in cells transfected with IMR-32-adapted JCVs was significantly greater than in those transfected with Mad-1 or CY. DNA replication assay and viral load verified that the IMR-32-adapted JCVs were replication-competent in 293cells, but not Mad-1 or CY JCVs. These results suggest that a 293 culture system with IMR-32-adapted JCVs may be a useful tool for assessing replication of JCV in vitro.WILEY-BLACKWELL, 2015年04月, MICROBIOLOGY AND IMMUNOLOGY, 59(4) (4), 238 - 242, 英語[査読有り]研究論文（学術雑誌）
- Although HIV-1 drug resistance is a major obstacle in Indonesia, information on drug resistance is limited. In this study, the viral subtype and appearance of drug resistance mutations in the HIV-1 protease (PR) and reverse transcriptase (RT) genes were determined among drug-treated, HIV-1-infected patients in Surabaya. HIV-1 patients who received antiretroviral therapy (ART) more than 2 years were randomly recruited regardless of the viral load or ART failure. Fifty-eight HIV-1 PR genes and 53 RT genes were sequenced. CRF01_AE viruses were identified as the predominant strain. Major drug resistance mutations were not detected in the PR genes. In contrast, 37.7% (20/53) of the participants had one or more major drug resistance mutations in the RT genes, predominantly M184V (28.3%), K103N (11.3%), and thymidine analogue mutations (TAMs) (20.8%). The high prevalence of drug resistance mutations in RT genes indicated the necessity of monitoring the effectiveness of ART in Indonesia.MARY ANN LIEBERT, INC, 2015年02月, AIDS RESEARCH AND HUMAN RETROVIRUSES, 31(2) (2), 255 - 259, 英語[査読有り]研究論文（学術雑誌）
- Background: The emergence of transmitted drug resistance (TDR) compromises the effect of antiretroviral therapy (ART), resulting in treatment failure of human immunodeficiency virus (HIV) disease. Although more than a decade has passed since ART was introduced into Indonesia, information on TDR is limited. Here, a genotypic study of TDR among ART-naive individuals was conducted in Surabaya, Indonesia. Method: HIV-1 seropositive participants were recruited from the communities of commercial sex workers and intravenous drug users as well as from the university teaching hospital in Surabaya. Protease (PR) and reverse transcriptase (RT) genes were sequenced in order to conduct HIV-1 subtyping and phylogenetic analysis and to detect TDR. TDR was defined as the presence of at least one surveillance drug resistance mutation on the WHO list or major drug resistance mutations in the International AIDS Society-USA panel. Result: Fifty two and 47 of the PR and RT genes, respectively, were successfully sequenced in the 58 samples. HIV-1 subtyping revealed that 86.3% (50/58) of the sequenced samples were classified as CRF01_AE, 8.6% as subtype B, 3.4% as B/CRF01_AE, and 1.7% as A/G/CRF01_AE. TDR of PR inhibitors was not detected in this study. In contrast, TDR of RT inhibitors was detected in 4.3% (2/47) of samples. In addition, minor drug resistance mutations were detected in 98.1% (51/52) and 12.8% (6/47) of PR and RT genes, respectively. Conclusion: This study clarified the predominance of the CRF01_AE strain in Surabaya, Indonesia. The prevalence of TDR was below 5%, indicating that the currently available first-line regimen is still effective in Surabaya. However, the prevalence might be underestimated since we detected only major population of HIV-1 in individuals. Therefore, continuous surveillance is required in order to detect the emergence of TDR in the early phase.BIOMED CENTRAL LTD, 2015年02月, AIDS research and therapy, 12, 5, 英語[査読有り]研究論文（学術雑誌）
- JC polyomavirus (JCV) causes progressive multifocal leukoencephalopathy (PML), a fatal demyelinating disease of the central nervous system (CNS) in immunocompromised patients, and particularly in the severe immunosuppression associated with acquired immunodeficiency syndrome (AIDS). HIV-1 can lead to the production of tumor necrosis factor-alpha (TNF-) in the CNS. Our aim was to examine the effects of TNF- on JCV gene expression and replication using a human neuroblastoma cell line, IMR-32, transfected with JCV DNA, M1-IMRb. Quantitative RT-PCR analysis of JCV large T antigen and VP1 mRNA, the viral DNA replication assay, and the DNase protection assay were carried out. TNF- treatment of IMR-32 cells transfected with JCV DNA induced large T antigen mRNA and JCV DNA replication, while other effects on VP1 mRNA expression and virus production were marginal. In addition, ELISA analysis of the nuclear p65 subunit of nuclear factor B (NF-B), which is a hallmark of NF-B pathway activation, of IMR-32 cells upon TNF- treatment showed that TNF- treatment activated the NF-B pathway in IMR-32 cells. Taken together, our results suggest that TNF- stimulation could induce JCV replication associated with the induction of JCV large T antigen mRNA through the NF-B pathway in IMR-32 cells transfected with JCV DNA. Our findings may contribute to further understanding of the pathogenesis of AIDS-related PML. J. Med. Virol. 86:2026-2032, 2014. (c) 2014 Wiley Periodicals, Inc.WILEY-BLACKWELL, 2014年12月, Journal of medical virology, 86(12) (12), 2026 - 2032, 英語[査読有り]研究論文（学術雑誌）
- Four serotypes of dengue virus (DENV-1 to DENV-4) and their genotypes are distributed in tropical and subtropical regions. Indonesia has been recently suggested as the origin of some dengue virus genotypes. In Surabaya, the second biggest city of Indonesia, we previously reported a shift of the predominantly circulating serotype from DENV-2 to DENV-1 in November 2008, followed by a genotype shift of DENV-1 from genotype IV (GIV) to genotype I (GI) in September 2009, based on nucleotide sequences in the envelope protein coding region. Since then, GI strains had predominantly circulated until December 2010. In this report, we investigated further DENV-1 transitions in Surabaya during 2011-2013 in order to comprehend dengue dynamics during 2008-2013 in more detail. From January 2011 through December 2011, only GIV strains were isolated, indicating that a genotype shift again took place from GI to GIV. In January 2012, GI and GIV strains started co-circulating, which continued until June 2013. To further investigate this phenomenon, analysis was performed at a clade level. GI and GIV strains isolated in Surabaya formed four and three distinct clades, respectively. Concomitant with co-circulation, new clade strains appeared in both genotypes. In contrast, some previously circulating clades were not isolated during co-circulation, indicating clade shifts. Among our Surabaya isolates, nucleotide and amino acid differences in the E region were, respectively, 1.0-2.3% and 0.2-1.0% for GI isolates and 2.0-6.3% and 0.0-1.8% for GIV isolates. Several characteristic amino acid substitutions in the envelope ectodomain were observed in some clades. After July 2013, DENV-1 strains were not isolated and were replaced with DENV-2. This study showed that continuous shifts of more than one genotype resulted in their co-circulation and subsequent disappearance and suggested the relevance of clade replacement to genotype co-circulation and disappearance in Surabaya. (C) 2014 Elsevier B.V. All rights reserved.ELSEVIER SCIENCE BV, 2014年12月, INFECTION GENETICS AND EVOLUTION, 28, 48 - 54, 英語[査読有り]研究論文（学術雑誌）
- Although human immunodeficiency virus type 1 (HIV-1) infection causes serious health problems in Indonesia, information in regard to drug resistance is limited. We performed a genotypic study on HIV-1 integrase derived from drug-naive individuals in Surabaya, Indonesia. Sequencing analysis revealed that no primary mutations associated with drug resistance to integrase inhibitors were detected; however, secondary mutations, V72I, L74I/M, V165I, V201I, I203M, and S230N, were detected in more than 5% of samples. In addition, V201I was conserved among all samples. Most integrase genes were classified into CRF01_AE genes. Interestingly, 40% of the CRF01_AE genes had an unusual insertion in the C-terminus of integrase. These mutations and insertions were considered natural polymorphisms since these mutations coincided with previous reports, and integrase inhibitors have not been used in Indonesia. Our results indicated that further studies may be required to assess the impact of these mutations on integrase inhibitors prior to their introduction into Indonesia.MARY ANN LIEBERT, INC, 2014年05月, AIDS RESEARCH AND HUMAN RETROVIRUSES, 30(5) (5), 489 - 492, 英語[査読有り]研究論文（学術雑誌）
- NATL INST INFECTIOUS DISEASES, 2014年05月, JAPANESE JOURNAL OF INFECTIOUS DISEASES, 67(3) (3), 227 - 229, 英語Phylogenetic Analysis of Dengue Virus Type 3 Strains Primarily Isolated in 2013 from Surabaya, Indonesia[査読有り]研究論文（学術雑誌）
- Background: The CD4 binding site (CD4bs) of envelope glycoprotein (Env) gp120 is a functionally conserved, important target of anti-human immunodeficiency virus type 1 (HIV-1) neutralizing antibodies. Two neutralizing human monoclonal antibodies, IgG1 b12 (b12) and VRC01, are broadly reactive neutralizing antibodies which recognize conformational epitopes that overlap the CD4bs of Env gp120; however, many CRF01_AE viruses are resistant to neutralization mediated by these antibodies. We examined the mechanism underlying the b12 resistance of the viruses using CRF01_AE Env (AE-Env)-recombinant viruses in this study. Results: Our results showed that an amino acid substitution at position 185 in the V2 region of gp120 played a crucial role in regulating the b12 susceptibility of AE-Env-recombinant viruses by cooperating with 2 previously reported potential N-linked glycosylation (PNLG) sites at positions 186 (N186) and 197 (N197) in the V2 and C2 regions of Env gp120. The amino acid residue at position 185 and 2 PNLG sites were responsible for the b12 resistance of 21 of 23 (> 91%) AE-Env clones tested. Namely, the introduction of aspartic acid at position 185 (D185) conferred b12 susceptibility of 12 resistant AE-Env clones in the absence of N186 and/or N197, while the introduction of glycine at position 185 (G185) reduced the b12 susceptibility of 9 susceptible AE-Env clones in the absence of N186 and/or N197. In addition, these amino acid mutations altered the VRC01 susceptibility of many AE-Env clones. Conclusions: We propose that the V2 and C2 regions of AE-Env gp120 contain the major determinants of viral resistance to CD4bs antibodies. CRF01_AE is a major circulating recombinant form of HIV-1 prevalent in Southeast Asia. Our data may provide important information to understand the molecular mechanism regulating the neutralization susceptibility of CRF01_AE viruses to CD4bs antibodies.BIOMED CENTRAL LTD, 2014年04月, RETROVIROLOGY, 11, 32, 英語[査読有り]研究論文（学術雑誌）
- Neutralizing antibodies induced by dengue virus (DENV) infection show viral infection-enhancing activities at sub-neutralizing doses. On the other hand, preimmunity against Japanese encephalitis virus (JEV), a congener of DENV, does not increase the severity of DENV infection. Several studies have demonstrated that neutralizing epitopes in the genus Flavivirus are mainly located in domain III (DIII) of the envelope (E) protein. In this study, chimeric premembrane and envelope (prM-E) gene-based expression plasmids of JEV and DENV1 with DIII substitution of each virus were constructed for use as DNA vaccines and their immunogenicity evaluated. Sera from C3H/He and ICR mice immunized with a chimeric gene containing DENV1 DIII on a JEV prM-E gene backbone showed high neutralizing antibody titers with less DENV infection-enhancing activity. Our results confirm the applicability of this approach as a new dengue vaccine development strategy.WILEY-BLACKWELL, 2014年02月, MICROBIOLOGY AND IMMUNOLOGY, 58(2) (2), 126 - 134, 英語[査読有り]研究論文（学術雑誌）
- Transmitted/founder virus is responsible for the establishment of human immunodeficiency virus type 1 (HIV-1) infection and induces primary anti-HIV-1 immune responses; therefore, it is important to study the viral population to understand the early events of HIV-1 infection. We amplified HIV-1 env genes from sera derived from recently infected Thai individuals, and established envelope glycoproteins (Env)-re-combinant viruses. Generated Env-recombinant viruses were tested for their neutralization susceptibility to neutralizing human monoclonal antibodies (NHMAbs) and entry inhibitors, as well as being subjected to genotypic analysis. Most recombinant viruses were susceptible to neutralization by NHMAbs to Env gp41, whereas approximately one-third of the recombinant viruses were susceptible to a NHMAb against the CD4 binding site of gp120. In addition, all env genes were classified into CRF01_AE genes and showed low genetic divergence. Taken together with our previous studies on CRF01_AE env genes derived from chronically infected Thai individuals, these results suggested that the immunological and genetic characteristics of CRF01_AE Env derived from recently infected Thai individuals were different from those derived from chronically infected individuals. (C) 2013 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.ELSEVIER SCIENCE BV, 2014年02月, MICROBES AND INFECTION, 16(2) (2), 142 - 152, 英語[査読有り]研究論文（学術雑誌）
- Background: Human immunodeficiency virus (HIV) infection and acquired immune deficiency syndrome (AIDS) cause serious health problems and have an impact on the Indonesian economy. In addition, the rapid epidemic growth of HIV is continuing in Indonesia. Commercial sex plays a significant role in the spread of HIV; therefore, in order to reveal the current HIV prevalence rate among commercial sex workers (CSWs), we conducted an epidemiological study on HIV infection among CSWs residing in Surabaya, the capital of East Java province of Indonesia with large communities of CSWs. Methodology/Principal Findings: The prevalence of HIV infection among 200 CSWs was studied. In addition, the subtype of HIV type 1 (HIV-1) and the prevalence of other blood-borne viruses, hepatitis B virus (HBV), hepatitis C virus (HCV) and GB virus C (GBV-C), were studied. The prevalence rates of HIV, hepatitis B core antibody, hepatitis B surface antigen, anti-HCV antibodies and anti-GBV-C antibodies were 11%, 64%, 4%, 0.5% and 0% among CSWs involved in this study, respectively. HIV-1 CRF01_AE viral gene fragments were detected in most HIV-positive samples. In addition, most CSWs showed low awareness of sexually transmitted diseases and had unprotected sex with their clients. Conclusions/Significance: The HIV prevalence rate among CSWs was significantly higher than that among the general population in Indonesia (0.2-0.4%). In addition, CSWs were at a high risk of exposure to HBV, although chronic HBV infection was less frequently established. Our results suggest the necessity of efficient prevention programs for HIV and other blood-borne viral infections among CSWs in Surabaya, Indonesia.PUBLIC LIBRARY SCIENCE, 2013年12月, PLOS ONE, 8(12) (12), e82645, 英語[査読有り]研究論文（学術雑誌）
- Universitas Airlangga, 2013年10月, Indonesian Journal of Tropical and Infectious Disease, 4(4) (4), 35 - 35研究論文（学術雑誌）
- Background: Development of a protective vaccine against human immunodeficiency virus type 1 (HIV-1) is an important subject in the field of medical sciences however, it has not yet been achieved. Potent and broadly neutralizing antibodies are found in the plasma of some HIV-1-infected patients, whereas such antibody responses have failed to be induced by currently used vaccine antigens. In order to develop effective vaccine antigens, it is important to reveal the molecular mechanism of how strong humoral immune responses are induced in infected patients. As part of such studies, we examined the correlation between the anti-HIV-1 neutralizing antibody response and disease progression. Methodology/Principal Findings: We evaluated the anti-HIV-1 neutralizing activity of plasma derived from 33 rapid and 34 slow progressors residing in northern Thailand. The level of neutralizing activity varied considerably among plasmas, and no statistically significant differences in the potency and breadth of neutralizing activities were observed overall between plasma derived from rapid and slow progressors however, plasma of 4 slow progressors showed neutralizing activity against all target viruses, whereas none of the plasma of rapid progressors showed such neutralizing activity. In addition, 21% and 9% of plasmas derived from slow and rapid progressors inhibited the replication of more than 80% of CRF01_AE Env-recombinant viruses tested, respectively. Neutralization of subtype B and C Env-recombinant viruses by the selected plasma was also examined however, these plasma samples inhibited the replication of only a few viruses tested. Conclusions/Significance: Although no statistically significant differences were observed in the potency and breadth of anti-HIV-1 neutralizing activities between plasma derived from rapid and slow progressors, several plasma samples derived from slow progressors neutralized CRF01_AE Env-recombinant viruses more frequently than those from rapid progressors. In addition, plasma derived from HIV-1-infected Thai patients showed CRF01_AE-specific neutralizing activity. © 2013 Sapsutthipas et al.2013年01月, PLoS ONE, 8(1) (1), e53920, 英語[査読有り]研究論文（学術雑誌）
- Cyclophilin A has attracted attention recently as a new target of anti-human immunodeficiency virus type 1 (HIV-1) drugs. However, so far no drug against HIV-1 infection exhibiting this mechanism of action has been approved. To identify new potent candidates for inhibitors, we performed in silico screening of a commercial database of more than 1,300 drug-like compounds by using receptor-based docking studies. The candidates selected from docking studies were subsequently tested using biological assays to assess anti-HIV activities. As a result, two compounds were identified as the most active. Specifically, both exhibited anti-HIV activity against viral replication at a low concentration and relatively low cytotoxicity at the effective concentration inhibiting viral growth by 50 %. Further modification of these molecules may lead to the elucidation of potent inhibitors of HIV-1.SPRINGER, 2013年01月, JOURNAL OF MOLECULAR MODELING, 19(1) (1), 465 - 475, 英語[査読有り]研究論文（学術雑誌）
- The incidence of progressive multifocal leukoencephalopathy (PML) has increased due to the AIDS pandemic, hematological malignancies, and immunosuppressive therapies. Recently, the number of cases of monoclonal antibody-associated PML has increased in patients treated with immunomodulatory drugs such as natalizumab. However, no common consensus regarding PML therapy has been reached in clinical studies. In order to examine the suppression of JC virus (JCV) replication by 3-aminobenzamide (3-AB), a representative PARP-1 inhibitor, a DNA replication assay was carried out using the neuroblastoma cell line IMR-32 and IMR-adapted JCV. The suppression of JCV propagation by 3-AB was also examined using JCI cells, which are a carrier culture producing continuously high JCV titers. The results indicated that PARP-1 inhibitors, such as 3-aminobenzamide (3-AB), suppress JCV replication and propagation significantly in vitro, as judged by DNA replication assay, hemagglutination, and real-time PCR analysis. It has been also shown that 3-AB reduced PARP-1 activity in IMR-32 cells. According to the results of the MTT assay, the enzyme activity of 3-AB-treated cells was slightly lower than that of DMSO-treated cells. However, the significant suppression of JCV propagation is not related to the slight decrease in cell growth. To our knowledge, this is the first report that PARP-1 inhibitor suppresses the replication of JCV significantly in neuroblastoma cell lines via the reduction of PARP-1 activity. Thus, PARP-1 inhibitors also may be a novel therapeutic drug for PML.2013年01月, Journal of medical virology, 85(1) (1), 132 - 7, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Variables influencing anti-human immunodeficiency virus type 1 neutralizing human monoclonal antibody (NhMAb) production among infected ThaisWe conducted this study to determine the clinical variables associated with the production of human immunodeficiency virus type 1 (HIV-1) circulating recombinant form (CRF) 01_AE neutralizing human monoclonal antibodies (NhMAbs) using a hybridoma technique. This cross sectional study was performed in 20 asymptomatic HIV-1-infected Thais. Peripheral blood mononuclear cells (PBMCs) were obtained from each study participant and fused with SPYMEG cells. Culture supernatant collected from growing hybridomas was tested for neutralizing activity against HIV-1 CRF01_AE Env-recombinant viruses. Fifty hybridomas expressing anti-HIV-1 NhMAbs with strong neutralizing activity against at least 1 CRF01_AE Env-recombinant virus were found. A positive association between the numbers of hybridomas produced and the CD4 counts of study participants (p = 0.019) was observed. NhMAb-producing hybridomas with strong neutralizing activity were mostly found in participants diagnosed with HIV-1 infection within the previous 1 year. The HIV-1 viral load was not significantly correlated with the numbers of either established hybridomas or clones expressing anti-HIV-1 NhMAbs with strong neutralizing activity. To our knowledge, this is the first study of NhMAb-producing hybridomas obtained from HIV-1 CRF01_AE-infected populations identified by antibody binding to HIV-1 V3 loop peptide enzyme-linked immunosorbent assay (ELISA) or TRUGENE HIV-1 Genotyping Assay (HIV-1 pol sequence). It provides important criterion to select study participants with high CD4 counts who produce large numbers of hybridoma clones. The results are valuable for further studies related to neutralizing antibodies production and HIV-1 vaccine development.SEAMEO TROPMED Network, 2013年, Southeast Asian Journal of Tropical Medicine and Public Health, 44(5) (5), 825 - 841, 英語[査読有り]研究論文（学術雑誌）
- Background: Double-stranded RNA (dsRNA) and its mimic, polyinosinic acid: polycytidylic acid [Poly (I:C)], are recognized by toll-like receptor 3 (TLR3) and induce interferon (IFN)-beta in many cell types. Poly (I:C) is the most potent IFN inducer. In in vivo mouse studies, intraperitoneal injection of Poly (I:C) elicited IFN-alpha/beta production and natural killer (NK) cells activation. The TLR3 pathway is suggested to contribute to innate immune responses against many viruses, including influenza virus, respiratory syncytial virus, herpes simplex virus 2, and murine cytomegalovirus. In Chikungunya virus (CHIKV) infection, the viruses are cleared within 7-10 days postinfection before adaptive immune responses emerge. The innate immune response is important for CHIKV clearance. Results: The effects of Poly (I:C) on the replication of CHIKV in human bronchial epithelial cells, BEAS-2B, were studied. Poly (I:C) suppressed cytopathic effects (CPE) induced by CHIKV infection in BEAS-2B cells in the presence of Poly (I:C) and inhibited the replication of CHIKV in the cells. The virus titers of Poly (I:C)-treated cells were much lower compared with those of untreated cells. CHIKV infection and Poly (I:C) treatment of BEAS-2B cells induced the production of IFN-beta and increased the expression of anti-viral genes, including IFN-alpha, IFN-beta, MxA, and OAS. Both Poly (I:C) and CHIKV infection upregulate the expression of TLR3 in BEAS-2B cells. Conclusions: CHIKV is sensitive to innate immune response induced by Poly (I:C). The inhibition of CHIKV replication by Poly (I:C) may be through the induction of TLR3, which triggers the production of IFNs and other anti-viral genes. The innate immune response is important to clear CHIKV in infected cells.BIOMED CENTRAL LTD, 2012年06月, VIROLOGY JOURNAL, 9, 114, 英語[査読有り]研究論文（学術雑誌）
- The high incidence of progressive multifocal leukoencephalopathy (PML) among individuals with acquired immunodeficiency syndrome (AIDS) is similar to the incidence of other immunocompromised diseases. The pathogenic JC virus (JCV) with rearranged regulatory regions (PML-type) causes PML, a demyelinating disease in the brains of immunocompromised patients. In a previous study, Tat protein, encoded by human immunodeficiency virus type 1 (HIV-1), markedly enhanced the expression of a reporter gene under control of the JCV late promoter. In order to examine the enhancement of JCV replication by Tat protein, the neuroblastoma cell line IMR-32 was used because it enables IMR-32-adapted JCV. The extent of JCV replication in IMR-32 cells treated with Tat protein was significantly higher than that in untreated IMR-32 cells. The enhancement of JCV propagation by Tat protein was also examined using IMR-32-derived JCV producing (JCI) cells which continuously produce JCV. Treatment of JCI cells with Tat protein led to a significant increase in the titers of progeny viruses. It has also been shown that Tat protein leads to a decrease in the expression of purine-rich element binding protein a (Pura) as an important mediator of JCV replication in IMR-32 cells. Thus, it is probable that Tat protein enhances JCV replication in IMR-32 cells via the down-regulation of Pura expression and cell proliferation. To our knowledge, this is the first report that exogenous Tat protein enhances the replication of JCV efficiently in neuroblastoma cell lines. J. Med. Virol. 84:555561, 2012. (C) 2011 Wiley Periodicals, Inc.WILEY-BLACKWELL, 2012年04月, JOURNAL OF MEDICAL VIROLOGY, 84(4) (4), 555 - 561, 英語[査読有り]研究論文（学術雑誌）
- Background: The envelope glycoproteins (Env), gp120 and gp41, are the most variable proteins of human immunodeficiency virus type 1 (HIV-1), and are the major targets of humoral immune responses against HIV-1. A circulating recombinant form of HIV-1, CRF01_AE, is prevalent throughout Southeast Asia; however, only limited information regarding the immunological characteristics of CRF01_AE Env is currently available. In this study, we attempted to examine the evolutionary pattern of CRF01_AE Env under the selection pressure of host immune responses. Methodology/Principal Findings: Peripheral blood samples were collected periodically over 3 years from 15 HIV-1-infected individuals residing in northern Thailand, and amplified env genes from the samples were subjected to computational analysis. The V5 region of gp120 showed highest variability in several samples over 3 years, whereas the V1/V2 and/or V4 regions of gp120 also showed high variability in many samples. In addition, the N-terminal part of the C3 region of gp120 showed highest amino acid diversity among the conserved regions of gp120. Chronological changes in the numbers of amino acid residues in gp120 variable regions and potential N-linked glycosylation (PNLG) sites are involved in increasing the variability of Env gp120. Furthermore, the C3 region contained several amino acid residues potentially under positive selection, and APOBEC3 family protein-mediated G to A mutations were frequently detected in such residues. Conclusions/Significance: Several factors, including amino acid substitutions particularly in gp120 C3 and V5 regions as well as changes in the number of PNLG sites and in the length of gp120 variable regions, were revealed to be involved in the molecular evolution of CRF01_AE Env. In addition, a similar tendency was observed between CRF01_AE and subtype C Env with regard to the amino acid variation of gp120 V3 and C3 regions. These results may provide important information for understanding the immunological characteristics of CRF01_AE Env.PUBLIC LIBRARY SCIENCE, 2011年11月, PLOS ONE, 6(11) (11), e27098, 英語[査読有り]研究論文（学術雑誌）
- VIRAL FACTORS INVOLVED IN ADAPTER-RELATED PROTEIN COMPLEX 2 ALPHA 1 SUBUNIT-MEDIATED REGULATION OF HUMAN IMMUNODEFICIENCY VIRUS TYPE 1 REPLICATIONThe presence of siRNA against adapter-related protein complex 2 alpha 1 subunit (AP2 alpha) enhances human immunodeficiency virus type 1 (HIV-1) replication by up-regulating nuclear transport of viral genome. In this report, we examined possible viral factors involved in AP2 alpha-mediated regulation of HIV-1 replication, namely, Gag matrix protein (MA), integrase (IN) and Vpr. Replication of mutant viruses lacking the nucleophilic property of one of these viral proteins was significantly enhanced by treating cells with AP2 alpha siRNA, indicating that Gag MA, IN or Vpr is not specifically involved in AP2 alpha-mediated enhancement of viral replication. In contrast, AP2 alpha siRNA showed no effect on the level of gene transduction mediated by HIV-1-derived lentiviral vector (LV). Although virus-like LV particle and parental HIV-1 particle are composed of almost equivalent viral structural proteins, LV particles lack three accessory proteins, Vif, Vpr and Vpu, and a large portion of the HIV-1 genome. Vif, Vpr and Vpu were dispensable for AP2 alpha siRNA-mediated enhancement of HIV-1 replication, indicating that a particular part of the HIV-1 genomic fragment deleted in the LV genome might be required for the enhancing effect of AP2 alpha siRNA on viral replication. Taken together, these results suggest that an as yet undetermined gene fragment of the HIV-1 genome is involved in AP2 alpha-mediated regulation of HIV-1 replication.SOUTHEAST ASIAN MINISTERS EDUC ORGANIZATION, 2011年03月, SOUTHEAST ASIAN JOURNAL OF TROPICAL MEDICINE AND PUBLIC HEALTH, 42(2) (2), 311 - 319, 英語[査読有り]研究論文（学術雑誌）
- CRF01_AE is a major subtype of human immunodeficiency virus type 1 (HIV-1) circulating in Southeast Asia, including Thailand. We performed genotypic studies on HIV-1 CRF01_AE integrase derived from plasma samples from drug-naive Thai patients. Direct sequencing of amplified CRF01_AE integrase genes revealed that although no primary mutations associated with drug resistance to integrase inhibitors were detected, at least one secondary mutation was found in 96% of samples. Our results indicate that the impact of these mutations on the baseline drug susceptibility of CRF01_AE viruses to integrase inhibitors may need to be addressed prior to the introduction of these drugs in Southeast Asian countries, including Thailand.MARY ANN LIEBERT INC, 2010年12月, AIDS RESEARCH AND HUMAN RETROVIRUSES, 26(12) (12), 1341 - 1343, 英語[査読有り]研究論文（学術雑誌）
- The high incidence of progressive multifocal leukoencephalopathy (PML) in AIDS patients compared with many other immunosuppressive diseases suggests that HIV-1 infection is strictly related to the activation of JC virus (JCV) propagation. In this report, propagation of PML-type JCV in COS-7-derived cell lines stably expressing HIV-1 Tat (COS-tat cells) has been examined. In COS-tat cells, production of viral particles and replication of genomic DNA were markedly increased compared to COS-7 cells, as judged by HA and real-time PCR analyses. These results demonstrate that COS-tat cells provide a useful model system for studying HIV-1 Tat-mediated propagation of PML-type JCV.WILEY-BLACKWELL, 2010年12月, MICROBIOLOGY AND IMMUNOLOGY, 54(12) (12), 758 - 762, 英語[査読有り]研究論文（学術雑誌）
- Recombinant human immunodeficiency virus type 1 (HIV-1) containing a CRF01_AE Gag, AE-Gag62, was significantly less susceptible to protease inhibitors (Pis) than the subtype B reference strain, NL4-3, therefore, the mechanism of how AE-Gag62 reduced viral drug susceptibility to Pls was studied in this report The results showed that the lysine residue at amino acid position 165 (K165) of AE-Gag62 played a role in reducing the drug susceptibility of the recombinant virus to Pls In addition, K165 potentially appears more frequently in CRF01_AE viruses than in the viruses of other major HIV-1 subtypes Although K165 had no effect on the extent of recombinant protease-mediated in vitro Gag cleavage, it enhanced the incorporation of the Gag-Pol precursor protein. p160, into virions Taken together, these results suggest that K165 of CRF01_AE Gag affects the regulation of virion assembly or maturation, and reduces viral drug susceptibility to Pls (C) 2010 Elsevier Inc All rights reserved.ACADEMIC PRESS INC ELSEVIER SCIENCE, 2010年09月, VIROLOGY, 405(1) (1), 129 - 138, 英語[査読有り]研究論文（学術雑誌）
- Two alpha-helical heptad repeats. N-HR and C-HR, located in the human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein gp41, play an important role in membrane fusion by forming a 6-helix bundle. C34, a peptide mimicking C-HR, inhibits the formation of the 6-helix bundle, thus, it has potential as a novel antiretroviral compound In order to improve the inhibitory effect of C34 on HIV-1 replication, we designed new C34-derived peptides based on computational analysis of the stable conformation of the 6-helix bundle Newly designed peptides showed a stronger inhibitory effect on the replication of recombinant viruses containing CRF01_AE, subtype B or subtype C Env than C34 or a fusion inhibitor, T-20 In addition, these peptides inhibited the replication of a T-20-resistant virus We propose that these peptides could be applied to develop novel antiretroviral compounds to inhibit the replication of various subtypes of HIV-I as well as of T-20-resistant variants (C) 2010 Elsevier Inc All rights reservedACADEMIC PRESS INC ELSEVIER SCIENCE, 2010年09月, VIROLOGY, 405(1) (1), 157 - 164, 英語[査読有り]研究論文（学術雑誌）
- Since December 1997, highly pathogenic avian influenza A H5N1 viruses have swept through poultry populations across Asian countries and been transmitted into African and European countries. We characterized 6 avian influenza H5N1 viruses isolated from humans in 2004 in Thailand. A highly pathogenic (HP) KAN353 strain showed faster replication and higher virulence in embryonated eggs compared to other strains, especially compared to the low pathogenic (LP) SP83 strain. HP KAN353 also showed strong cytopathogenicity compared to SP83 in Madin-Darby canine kidney cells. Interestingly, LP SP83 induced smaller plaques compared to other strains, especially HP KAN353. PB2 amino acid 627E may contribute to low virulence, whereas either PB2 amino acid 627 K or the combination of 627E/701N seems to be associated with high virulence. The in vitro assays used in this study may provide the basis for assessing the pathogenesis of influenza H5N1 viruses in vivo.BIOMED CENTRAL LTD, 2010年06月, VIROLOGY JOURNAL, 7, 112, 英語[査読有り]研究論文（学術雑誌）
- A recombinant human monoclonal antibody, IgG1 b12 (b12), recognizes a conformational epitope on human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein (Env) gp120 that overlaps the CD4 binding domain. Although b12 is able to broadly neutralize HIV-1 subtype B, C, and D viruses, many HIV-1 CRF01_AE viruses are resistant to b12-mediated neutralization. In this report, we examined the molecular mechanisms underlying the low neutralization susceptibility of CRF01_AE viruses to b12, using recently established CRF01_AE Env recombinant viruses. Our results showed that two potential N-linked glycosylation (PNLG) sites in the V2 and C2 regions of Env gp120 played an important role in regulating the susceptibility of CRF01_AE Env to b12. The locations of these PNLG sites correspond to amino acid positions 186 and 197 in HXB2 Env gp120; thus, they are designated N186 and N197 in this study. Removal of N186 significantly conferred the b12 susceptibility of 2 resistant CRF01_AE Env clones, 65CC4 and 107CC2, while the introduction of N186 reduced the b12 susceptibility of a susceptible CRF01_AE Env clone, 65CC1. In addition, removal of both N186 and N197 conferred the b12 susceptibility of 3 resistant CRF01_AE Env clones, 45PB1, 62PL1, and 101PL1, whereas the removal of either N186 or N197 was not sufficient to confer the b12 susceptibility of these CRF01_AE Env clones. Finally, removal of N197 conferred the b12 susceptibility of 2 resistant CRF01_AE Env clones lacking N186, 55PL1 and 102CC2. Taken together, we propose that two PNLG sites, N186 and N197, in Env gp120 are important determinants of the b12 resistance of CRF01_AE viruses.AMER SOC MICROBIOLOGY, 2010年05月, JOURNAL OF VIROLOGY, 84(9) (9), 4311 - 4320, 英語[査読有り]研究論文（学術雑誌）
- SUSTAINED APPEARANCE OF DRUG RESISTANCE-ASSOCIATED MUTATIONS IN HIV-1 CRF01_AE PROTEASE AND REVERSE TRANSCRIPTASE DERIVED FROM PROTEASE INHIBITOR-NAIVE THAI PATIENTSPrevious studies revealed that HIV-1 CRF01_AE viruses derived from antiretroviral drug-naive Thai patients contained several protease (PR) inhibitor (PI) resistance-associated mutations In this report, we examined the sustained appearance of drug resistance-associated mutations in CRF01_AE PR and reverse transcriptase (RT). Peripheral blood samples were collected every 3 months from April 2008 to April 2009 from 39 HIV-1-infected Thai patients, including 17 drug-naive and 22 RT. inhibitors (RTIs)-treated individuals, and polymerase chain reaction-mediated amplification and sequencing analysis of the viral genome encoding PR and RT were performed. We successfully analyzed the deduced amino acid sequence of CRF01_AE PR and RT derived from samples continuously collected from 15 drug-naive and 20 RTIs-treated patients. Drug resistance-associated mutations were continuously detected in CRF01_AE PR derived from most patients. The continuous appearance of such PR mutations was observed not only in the proviral DNA genome derived from peripheral blood mononuclear cells, but also in the viral RNA genome of plasma virus. In contrast, RTI resistance-associated mutations were only sporadically detected in samples derived from drug-naive and RTIs-treated patients, except for the continuous appearance of two mutations in samples derived from two drug-naive patients. Our results demonstrate that many PI resistance-associated mutations and only a few RTI resistance-associated mutations continuously appear in CRF01_AE viruses derived from PI-naive patients residing in northern ThailandSOUTHEAST ASIAN MINISTERS EDUC ORGANIZATION, 2010年03月, SOUTHEAST ASIAN JOURNAL OF TROPICAL MEDICINE AND PUBLIC HEALTH, 41(2) (2), 347 - 357, 英語[査読有り]研究論文（学術雑誌）
- We report 25 env gp160 sequences from patients in three geographically distinct districts of Thailand, i.e., Lampang in the north, Trang in the south and Rayong in the east. One of these is a CRF01_AE/subtype B recombinant and the other 24 sequences are purely CRF01_AE. Very little interpopulation diversity was observed between the sequences from the three different geographic regions and from those previously reported by our laboratory from central Thailand. Potential N-linked glycosylation sites (PNLGs) were reasonably conserved among the 25 sequences: we found 15 highly conserved PNLGs on gp120 and 4 almost fully conserved PNLGs on gp41. Analysis of coreceptor tropism revealed that six of the isolates were dual tropic and the others were R5 tropic. We also examined a rare seven amino acid deletion found in one isolate at position 847-853 on gp41. These results may enhance our understanding of HIV-1 currently circulating in Thailand.MARY ANN LIEBERT INC, 2010年02月, AIDS RESEARCH AND HUMAN RETROVIRUSES, 26(2) (2), 223 - 227, 英語[査読有り]研究論文（学術雑誌）
- The distribution of the G type of human rotavirus was surveyed in Thailand between July 1993 and June 2007. A significant yearly change in the distribution of the G type distribution was found. From 1993-1994 to 1998-1999, the G1 type was the most dominant. In 1999-2000, G9 began to appear at a high frequency. In 2000-2001, 2001-2002, and 2002-2003, G9 was very common. In 2003-2004, G1 became the most prevalent type again, and since then it has been detected at the highest frequency. G12 strains, which were first detected in 1998-1999,were also found in 2004-2005 and 2006-2007. The G4 and G3 types were moderately prevalent in 2001-2002 and 2004-2005, respectively. Nucleotide sequence analysis of the VP7 genes of the G9 and G12 strains which reemerged in Thailand showed that they were each similar to the contemporary strains in other countries. J. Med. Virol. 82:157-163, 2010. (C) 2009 Wiley-Liss, Inc.WILEY-LISS, 2010年01月, JOURNAL OF MEDICAL VIROLOGY, 82(1) (1), 157 - 163, 英語[査読有り]研究論文（学術雑誌）
- Pathogenic JCV with rearranged regulatory regions (PML-type) causes PML, a demyelinating disease, in the brains of immunocompromised patients. On the other hand, archetype JCV persistently infecting the kidney is thought to be converted to PML-type virus during JCV replication in the infected host under immunosuppressed conditions. In addition, Tat protein, encoded by HIV-1, markedly enhances the expression of a reporter gene under control of the JCV late promoter. In order to examine the influence of Tat on JCV propagation, we used kidney-derived COS-7 cells, which only permit archetype JCV, and established COS-tat cells, which express HIV-1 Tat stably. We found that the extent of archetype JCV propagation in COS-tat cells is significantly greater than in COS-7 cells. On the other hand, COS-7 cells express SV40 T antigen, which is a strong stimulator of archetype JCV replication. The expression of SV40 T antigen was enhanced by HIV-1 Tat slightly according to real-time RT-PCR, this was not closely related to JCV replication in COS-tat cells. The efficiency of JCV propagation depended on the extent of expression of functional Tat. To our knowledge, this is the first report of increased production of archetype JCV in a culture system using cell lines stably expressing HIV-1 Tat. We propose here that COS-tat cells are a useful tool for studying the role of Tat in archetype JCV replication in the development of PML.WILEY-BLACKWELL PUBLISHING, INC, 2009年11月, MICROBIOLOGY AND IMMUNOLOGY, 53(11) (11), 621 - 628, 英語[査読有り]研究論文（学術雑誌）
- Hepatitis E virus (HEV) infection in pigs was investigated in two principal swine farming areas in Thailand. Anti-HEV antibodies and HEV RNA in sera were examined in 258 pigs reared on five commercial farms from age 1 to 6.5 months and sows. Overall, 167 of 258 (64.7%) pigs were positive for anti-HEV IgG, while 20 of 258 (7.75%) had detectable HEV RNA. Sequence analysis of 20 HEV isolates obtained from viremic pigs revealed that they were 92.3-100% identical to each other and had 82.2-88.2% nucleotide similarity to other reported genotype 3 isolates in 415 nucleotide sequences within ORF2 region. Further characterization by sequencing the complete genome of the Thai swine HEV isolate (named Thai-swHEV07) and phylogenetic analysis showed that Thai-swHEV07 segregated into a cluster consisting of swine isolates from Japan, Mongolia, and Kyrgyzstan within the HEV genotype 3. The Thai-swHEV07 had a genomic length of 7,229 nt excluding the polyadenylated region at 3' terminus of the genome. Comparison of Thai-swHEV07 and 27 reported strains of genotype 3 revealed 80.4-85.9% nucleotide identity, with the highest identity of 85.9% to the novel swHEV strain from Mongolia. These findings suggest that genotype 3 HEV isolates are markedly heterogeneous. J. Med. ViroL 81:657-664, 2009. (C) 2009 Wiley-Liss, Inc.WILEY-LISS, 2009年04月, JOURNAL OF MEDICAL VIROLOGY, 81(4) (4), 657 - 664, 英語[査読有り]研究論文（学術雑誌）
- Superinfection rates of human immunodeficiency virus type 1 (HIV-1) have increasingly been leading to more variation in HIV-1, as evidenced by the emergence of circulating recombinant forms (CRFs). We recently reported complementation in a persistently replication-defective subtype B-infected cell clone, L-2, by superinfection with CRF15_01B. The L-2 cells continuously produce immature particles due to a one-base insertion at pol protease. Proviruses in the superinfected cells carried both subtypes and produced particles with a mature morphology. In this study, we examined possible recombination following complementation to generate replication-competent variants by using three cell clones prepared from superinfected L-2 cells. The individual clones predominantly expressed the initial subtype B-derived mature Gag proteins. However, the viral particles carried both subtype B with the mutation and wild-type CRF15_01B at pol, suggesting the generation of virions with heterozygous RNAs. Interestingly, with cell-free passages of the progeny, defective particles disappeared, and were replaced with heterogeneous recombinants in the pol region with sequences derived from CRF15_01B that expressed subtype B phenotype. Thus, even a defective form of persistent HIV-1 can become replication-competent through superinfection-mediated complementation followed by recombination. These findings suggest the significance of long-lived infected cells as recipients for superinfection. (C) 2008 Elsevier Masson SAS. All rights reserved.ELSEVIER SCIENCE BV, 2008年04月, MICROBES AND INFECTION, 10(5) (5), 504 - 513, 英語[査読有り]研究論文（学術雑誌）
- CXCR4-using HIV-1 was previously shown to replicate more efficiently in a healthy donor-derived CD4(+) CD38(+) than in a CD4+ CD38(-) T-cell subset after stimulation with interleukin(IL)-4. Here, we identified 3 cellular genes, which were expressed to a higher level in an IL-4-stimulated CD38(-) subset. One of the 3 genes, RNF125/TRAC-1, was involved in the down-regulation of HIV-1 replication not only in cell lines, but also in peripheral blood mononuclear cells. RNFI25/TRAC-1 bears the RING finger domain, important for E3 ubiquitin protein ligase. Mutations in this domain of RNFl25/TRAC-1 led to the loss of HIV-1 down-modulatory activity, suggesting that E3 ligase activity is necessary. In addition, the results of Northern blotting and reporter gene analysis indicated that RNFl25/TRAC-1 function occurs at the viral transcription step. These results suggest that RNFI25/TRAC-1 could function to recruit host factor(s) controlling HIV-1 transcription to the ubiquitin-proteasome pathway. (C) 2007 Elsevier Inc. All rights reserved.ACADEMIC PRESS INC ELSEVIER SCIENCE, 2007年11月, VIROLOGY, 368(1) (1), 191 - 204, 英語[査読有り]研究論文（学術雑誌）
- RNA interference (RNAi) has become one of the most powerful and popular approach on gene silencing in clinical research study especially in virology due to the gene-specific suppression property of small interfering RNA (siRNA). In this report, we demonstrate that expression of vector-mediated small hairpin RNA (shRNA) against human immunodeficiency virus type 1 (HIV-1) integrase (IN), one of the three important enzymes in HIV infection by controlling the integration of viral RNA to host DNA, could suppress the protein synthesis of EGFP-tagged IN in HeLa cell model efficiently. Furthermore, we show that IN shRNA can successfully reduce the HIV particles production in 293T cells at the level similar to the positive control of HIV-1 tat shRNA. These results provide the therapeutic possibility of HIV replication using RNAi against HIV-1 integrase. (C) 2007 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.ELSEVIER SCIENCE BV, 2007年07月, FEBS LETTERS, 581(17) (17), 3253 - 3259, 英語[査読有り]研究論文（学術雑誌）
- Superinfection with human immunodeficiency virus type 1 (HIV-1) in human subjects, defined as reinfection with a heterologous strain of HIV-1, has become a topic of great interest. To illustrate the significance of this occurrence, we performed HIV-1 superinfection of L-2 cells, which were isolated from MT-4 cells persistently infected with subtype B HIV-1 as a cell clone continuously producing defective HIV-1 particles. L-2 cells carrying provirus with a one-base insertion in the pol protease were superinfected with HIV-1 derived from primary isolates of subtype B or CRF01_AE. The kinetics of the superinfection in L-2 were very slow compared with those of primary infections in MT-4. Interestingly, L-2 shifted after superinfection to become a producer of highly cytopathogenic HIV-1. Molecular characterization revealed that superinfection occurred in only about 10% of the CRF01_AE-superinfected L-2, which carried provirus of both subtypes and produced viral particles containing genomic RNA of both subtypes. Surprisingly, such cytopathogenic HIV-1 showed predominantly the original subtype B phenotype. Thus, the mechanism of the production of cytopathic HIV-1 seemed to be mediated by trans complementation with pol products of superinfected CRF01_AE. These findings suggest the significance of long-lived infected cells as recipients for superinfection that could result in the generation of new HIV-1 variants with high virulence in patients who are off therapy or do not adhere to treatment, and may indicate the need for precautions against such superinfection. (c) 2006 Elsevier SAS. All rights reserved.ELSEVIER SCIENCE BV, 2006年06月, MICROBES AND INFECTION, 8(7) (7), 1773 - 1782, 英語[査読有り]研究論文（学術雑誌）
- Transactivation domain (TAD)-truncated p73, Delta Np73, associates with p53, resulting in suppression of p53's functions. Using p53 null cell lines, we examined whether or not Delta Np73 can regulate gene expression in a p53-independent manner. When Delta Np73 alpha was cotransfected with a luciferase reporter plasmid with various enhancer elements, NF kappa B-responsive luciferase gene expression was selectively up-regulated by Delta Np73 alpha, but not by other p73-isoforms with TAD and Delta Np73 beta. Deletion of the TAD endowed p73 alpha with the ability to enhance the responsive gene's expression, but deletion of the N-terminal proline-rich domain (PRD) rendered the TAD-deleted p73 alpha inactive. Considering the inability of Delta Np73 beta, which is the C-terminus-truncated form of Delta Np73 alpha, to function, these results indicate that both the PRD and C-terminus are necessary for Delta Np73 alpha to can activate NF kappa B-responsive luciferase expression. Over-expression of p53 suppressed the TAD-truncated p73 alpha-mediated luciferase expression, suggesting that p53 interferes with the TAD-truncated p73 alpha-mediated activation of NF kappa B. Inhibitors for NF kappa B activation reduced the TAD-truncated p73 alpha-dependent NF kappa B-responsive gene expression, indicating that TAD-truncated p73 alpha activates NF kappa B as does TNF alpha. in addition to the results obtained in the reporter gene assay, TAD-truncated p73a stimulated the translocation of NF kappa B to the nucleus and the expression of an endogenous NF kappa B-responsive gene, Bcl-XL. Taken together, these results demonstrate that TAD-truncated p73 alpha can activate NF kappa B. (c) 2005 Elsevier Inc. All rights reserved.ELSEVIER INC, 2006年05月, EXPERIMENTAL CELL RESEARCH, 312(8) (8), 1254 - 1264, 英語[査読有り]研究論文（学術雑誌）
- This study examined the efficiency of human immunodeficiency virus type 1 (HIV-1) integration in poly(ADP-ribose)polymerase-1 (PARP-1)-deficient murine cells and in human cell lines transfected with small interfering RNA against PARP-1 (PARP-1 siRNA). To semi-quantify the amount of integrated HIV-1 genome, real-time nested PCR was carried out using primers specific for Alu and alphoid DNA combined with primers for the HIV-1 genome. The results showed that the integration efficiency of the HIV-1 genome near Alu DNA, which is randomly distributed in the chromosome, is reduced in PARP-1-deficient murine cells, but not in PARP-1 siRNA-transfected human cells. By contrast, the integration efficiency of the HIV-1 gnome near alphoid DNA, which is localized in the centromere region, is significantly reduced in PARP-1-deficient murine cells and in PARP-1 siRNA-transfected human cells. These results suggest that PARP-1 is required for HIV-1 integration near the centromere region both in human and murine cells. (c) 2005 Elsevier Inc. All rights reserved.ACADEMIC PRESS INC ELSEVIER SCIENCE, 2005年08月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 334(2) (2), 412 - 417, 英語[査読有り]研究論文（学術雑誌）
- The capacity of human immunodeficiency virus type 1 (HIV-1) to infect resting cells and to produce progeny particles may contribute significantly to its pathogenicity in vivo. We previously reported that primary culture of resting CD4(+) CD38(+) T-lymphocyte subset had higher production rate of CXCR4-using (X4) HIV-1 than CD4(+) CD38(-) subset. Interleukin (IL)-4 highly contributed to the up-regulation of the X4 virus production in the CD38(+) subset. Here, we show evidences that IL-4 treatment of both resting CD38(+) and CD38- subsets allowed the adsorption, entry, and integration of X4 virus at similar rates, while the following viral transcription rate was significantly lower in the CD38(-) than CD38(+) subset. Treatment of the CD38 subsets with IL-4 or phytohemagglutinin revealed no association of X4 virus replication ability in the subsets with classic T-cell activation or proliferation. Interestingly, the activator protein (AP)-1 was significantly activated in the CD38(+) subset after IL-4 treatment, while both nuclear factor (NF)-kappaB and signal transducers and activator of transcription (STAT)-6 were activated in the IL-4-treated CD38(+) and CD38(-) subsets at similar levels. Thus, IL-4-dependent X4 HIV-1 transcription occurs efficiently in the CD38(+) but not CD38- subset of CD4(+) population and AP-1 could play a significant role on viral transcription, leading to the up-regulated X4 virus production in the CD38(+) subset.CENTER ACADEMIC PUBL JAPAN, 2005年, MICROBIOLOGY AND IMMUNOLOGY, 49(2) (2), 155 - 165, 英語[査読有り]研究論文（学術雑誌）
- We established small interfering RNA (siRNA) directed against poly(ADP-ribose) polymerase 1 (PARP-1) that effectively reduces the expression of PARP-1 in two human cell lines. Established siRNA against PARP-1 significantly suppressed human immunodeficiency virus type 1 (HIV-1) replication, as well as the activation of the integrated HIV-1 long terminal repeat promoter. These results indicate that PARP-1 is required for efficient HIV-1 replication in human cells. We propose that PARP-1 may serve as a cellular target for RNA interference-mediated gene silencing to inhibit HIV-1 replication.AMER SOC MICROBIOLOGY, 2004年08月, JOURNAL OF VIROLOGY, 78(16) (16), 8931 - 8934, 英語[査読有り]研究論文（学術雑誌）
- DNp73 is a transactivation domain (TAD)-truncated form of p73. The ability of DNp73alpha to regulate gene expression was examined using reporter assays with luciferase gene constructs. Among various promoter-regulated reporter genes tested, heat shock factor (HSF)-responsive gene expression was selectively activated by DNp73alpha, but not by other p73-isoforms with TAD and DNp73beta. Deletion of TAD endowed p73alpha with the ability to activate HSF-responsive gene expression, but deletion of N-terminal proline-rich domain (PRD) rendered both DNp73alpha and the TAD-deleted p73alpha inactive. Considering the inability of DNp73beta, which is the C-terminus-truncated form of DNp73alpha, to function, these results indicate that both the PRD and C-terminus are necessary for DNp73a to be able to activate the HSF-dependent gene expression. In addition to the reporter gene expression, both DNp73alpha, and TAD-deleted p73alpha activated the expression of an endogenons gene, hsp70, corresponding with an increase in the active form of HSF1. Taken together. these results demonstrate that TAD-truncated p73alpha can activate HSF-dependent gene expression via induction of active HSF1. (C) 2004 Elsevier Inc. All rights reserved.ACADEMIC PRESS INC ELSEVIER SCIENCE, 2004年05月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 317(3) (3), 865 - 872, 英語[査読有り]研究論文（学術雑誌）
- Both the full-length and spliced RNA species of HIV-1 possess the necessary cis-acting elements including the primer binding site (PBS), the polypurine tract (PPT), as well as the 5'R and 3'R regions that are needed for their conversion to double-stranded cDNA through reverse transcription. Since measurable amounts of spliced viral RNA molecules can be detected within virus particles, we have examined the potential for reverse transcription of such virion-associated spliced viral RNA upon infection of permissive cells. Analysis of viral cDNA species by PCR and DNA sequencing not only led to the identification of viral DNA molecules that were reverse transcribed from full-length viral RNA, but also DNA molecules that displayed the same nucleotide sequences as those found in spliced viral RNA, except that the former harbored the complete long terminal repeats (LTR), a feature that distinguishes proviral DNA from viral genomic RNA. Further studies revealed various types of cDNA species that resemble the spliced viral RNA encoding each of the env, tat, rev, or nef genes, of which the nef cDNA represents the majority. Therefore, spliced HIV-1 RNA molecules must have been reverse transcribed along with full-length viral RNA during infection.MARY ANN LIEBERT INC PUBL, 2004年02月, AIDS RESEARCH AND HUMAN RETROVIRUSES, 20(2) (2), 203 - 211, 英語[査読有り]研究論文（学術雑誌）
- NF-kappaB-dependent, as well as human immunodeficiency virus type-1 (HIV-1) long terminal repeat (LT R)-dependent, reporter gene expression was significantly impaired in cells derived from poly(ADP-ribose) polymerase-1 (PARP-1)-knockout (PARP-1 -/-) mice. In addition, the level of protein acetylation was markedly lower in PARP-1 -/- cells than control (PARP-1 +/+) cells. Surprisingly, the expression levels of historic acetyltransferases (HATs), p300, cAMP response element-binding protein-binding protein (CBP), and p300/CBP-associated factor (PCAF), were significantly reduced in PARP-1 -/- cells, as compared with PARP-1 +/+ cells. These results suggest that PARP-I is required for the proper expression of particular HATs. Since p300 and CBP are coactivators of NF-kappaB, we propose here that PARP-1 participates in NF-kappaB-dependent transcription by means of maintaining the expression of HATs. (C) 2003 Elsevier Inc. All rights reserved.ACADEMIC PRESS INC ELSEVIER SCIENCE, 2003年10月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 310(2) (2), 312 - 317, 英語[査読有り]研究論文（学術雑誌）
- The Tat protein of human immunodeficiency virus type 1 (HIV-1) has been shown to restrict premature reverse transcription at late stages of virus infection and to thus ensure the integrity of the viral RNA genome for packaging. To gain further insights into the roles of Tat in HIV-1 reverse transcription, we have assessed its effects on the first-strand transfer during the synthesis of minus-strand DNA through use of a reconstituted cell-free system. The results demonstrated that a form of Tat, containing only the first exon (Tat72), was able to enhance the first-strand transfer as efficiently as did the viral nucleocapsid protein. Coincidentally, this form of Tat was unable to inhibit the production of minus-strand strong-stop DNA. Further studies with various mutated forms of Tat showed that its Cys-rich region, rather than the core and Arg-rich domains, was essential for this strand transfer activity. Moreover, this activity of Tat is largely independent of the TAR RNA structure. Although full-length Tat protein (Tat86) was also able to promote strand transfer, this activity was limited by a strong overall inhibition of reverse transcription because of the presence of the second Tat exon. Other nucleic-acid-binding proteins (e.g., single-strand DNA-binding protein) were employed as negative controls and were unable to promote strand transfer in these assays. We propose that Tat possesses nucleic acid chaperone activity and can promote the first-strand transfer during HIV-1 reverse transcription; however, these activities are restricted by the second Tat exon, and the roles of these Tat activities in viral replication remain to be elucidated. (C) 2003 Elsevier Science (USA). All rights reserved.ACADEMIC PRESS INC ELSEVIER SCIENCE, 2003年03月, VIROLOGY, 307(1) (1), 154 - 163, 英語[査読有り]研究論文（学術雑誌）
- Human immunodeficiency virus type-1 Tat has been proposed to play a role in the regulation of reverse transcription. We previously demonstrated that wild-type Tat can augment viral infectivity by suppressing the reverse transcriptase (RT) reaction at late stages of the viral life cycle in order to prevent the premature synthesis of potentially deleterious viral DNA products. Here we have performed a detailed analysis of the cell-free reverse transcription reaction to elucidate the mechanism(s) whereby Tat can affect this process. Our results show that Tat can suppress nonspecific DNA elongation while moderately affecting the specific initiation stage of reverse transcription. In addition, Tat has an RNA-annealing activity and can promote the placement of tRNA onto viral RNA. This points to a functional homology between Tat and the viral nucleocapsid (NC) protein that is known to be directly involved in this process. Experiments using a series of mutant Tat proteins revealed that the cysteine-rich and core domains of Tat are responsible for suppression of DNA elongation, while each of the cysteine-rich, core, and basic domains, as well as a glutamine-rich region in the C-terminal domain, are important for the placement of tRNA onto the viral RNA genome. These results suggest that Tat can play at least two different roles in the RT reaction, i.e., suppression of DNA polymerization and placement of tRNA onto viral RNA. We believe that the first of these activities of Tat may contribute to the overall efficiency of reverse transcription of the viral genome during a new round of infection as well as to enhanced production of infectious viral particles. We hypothesize that the second activity, illustrating functional homology between Tat and NC, suggests a potential role for NC in the displacement of Tat during viral maturation.AMER SOC MICROBIOLOGY, 2002年04月, JOURNAL OF VIROLOGY, 76(8) (8), 3637 - 3645, 英語[査読有り]研究論文（学術雑誌）
- Brain and liver extracts of rats at different stages after birth were examined for cytochrome c/dATP-dependent caspase (DEVDase)-activation (mitochondria pathway) in vitro. The caspase-activating activity in the brain extracts rapidly decreased after birth, reaching approximately 50 and 5%, at I and 2 weeks, respectively, of that in a 3-days-newborn sample, and essentially no caspase-activation was detected in the adult rat brain extracts. Such a dramatic change was not detected in the liver samples, suggesting that the observed abrogation of the cytochrome c-dependent mitochondria pathway after birth is a brain-specific event. In order to determine the factor(s) lacking in adult brain, we separately measured Apaf-1, procaspase 9, and pro-DEVDase activities using a supplementation assay. In adult brain, Apaf-1 activity was scarcely detected, while the tissue retained low but significant amounts of procaspase 9 (16% of that in the fetal tissue) and a pro-DEVDase (3.4%). In contrast, adult liver extracts retained relatively high levels of all of these factors. Immunoblot analyses clearly indicated that the expression of Apaf-1 and procaspase 3 is markedly suppressed within 4 weeks after birth in brain tissue while they are even expressed in adult liver. Considering these results together, we propose that, in the brain, the cytochrome c-dependent mitochondria pathway, which is essential for the programmed cell death during normal morphogenesis, is abrogated within 2-4 weeks after birth, whereas the pathway is still active in other adult tissues such as liver.JAPANESE BIOCHEMICAL SOC, 2002年01月, JOURNAL OF BIOCHEMISTRY, 131(1) (1), 131 - 135, 英語[査読有り]研究論文（学術雑誌）
- We have examined the role of the human immunodeficiency virus type 1 (HIV-1) Tat protein in the regulation of reverse transcription. We show that a two exon but not a one-exon form of Tat markedly suppressed cell-free reverse transcriptase (RT) activity. Conversely, viruses expressing two-exon Tat (pNL43 and pNL101) showed rapid replication kinetics and more efficient endogenous RT activity compared with viruses expressing one-exon Tat (pM1ex). The pM1ex virions, as well as pM1ex-infected cells, also contained higher levels of viral DNA than did either the pNL43 or pNL101 viruses, indicating that reverse transcription might have continued during later stages of viral replication in the absence of the second Tat exon. Moreover, degradation of viral genomic RNA was more apparent in the pM1ex virions. Accordingly, we propose that the two-exon Tat may help augment viral infectivity by suppressing the reverse transcription reaction during late stages of viral synthesis and by preventing the synthesis of potentially deleterious viral DNA products.AMER SOC MICROBIOLOGY, 2001年03月, JOURNAL OF VIROLOGY, 75(6) (6), 2675 - 2683, 英語[査読有り]研究論文（学術雑誌）
- American Society for Biochemistry {\&} Molecular Biology ({ASBMB}), 2000年11月, Journal of Biological Chemistry, 276(9) (9), 6711 - 6719[査読有り]研究論文（学術雑誌）
- The DNA-binding activity of NF-KB in nuclear extracts of poly(ADP-ribose) polymerase (PARP)-defective mutant L1210 cell clones was markedly increased and was inversely correlated with the PARP content in these cells. The DNA-binding activity of NF-kappa B in a clone with the lowest PARP content (Cl-3527, contained 6 % of PARP of wild type cells) was about 35-fold of that of the wild-type cells, whereas the change in the DNA-binding activity of AP-1 and SP-1 in the mutant was relatively small or not so significant. Transfection of a PARP-expressing plasmid to the mutant cells decreased the abnormally high levels of NF-kappa B complexes, especially p50/p65(Rel A) complex, to near the normal level. Moreover, poly(ADP-ribosyl)ation of nuclear extracts in vitro suppressed the ability of NF-kappa B to form a complex with its specific DNA probe by approx. 80 Ob. Further analysis with purified recombinant NF-KB proteins revealed that both rp50 and rMBP-p65 (Rel A) proteins, but not rGST-I kappa B, could be poly(ADP-ribosyl)ated in vitro and that the modification resulted in a marked decrease in the DNA-binding activity of rMBP-p65, whereas a slight activation was observed in rp50. Poly(ADP-ribosyl)ated p65/NF-kappa B was detected in the cytosol of wild type L1210 cells by immunoblotting with anti-poly(ADP-ribose) and anti-p65 antibodies. Taken together, these results strongly suggest that PARP is involved in the regulation of NF-kappa B through the protein modification.Portland Press Ltd., 2000年03月, Biochemical Journal, 346(3) (3), 641 - 649, 英語[査読有り]研究論文（学術雑誌）
- Protease inhibitors are widely used in the treatment of human immunodeficiency virus type 1 (HIV-1)-infected individuals and show a drastic effect on the reduction of virus load. We previously reported that doughnut-shaped, protease-defective gp120-containing HIV-1 particles from an L-2 cell clone, carrying a provirus with mutations at the pol (protease), env (gp41) and nef genes, rapidly and more effectively induces virus particle-mediated syncytia formation of uninfected T-cells, than a parental wild-type laboratory strain of HIV-1 (LAI). In this study, we examined the possibility of whether enhanced syncytia formation is mediated by morphologically similar doughnut-shaped particles obtained after treatment of LAI-infected cells with the protease inhibitors L-689,502, DMP-323, RO-31-8959, and KNI-272. Utilizing such protease inhibitor-induced particles and a clone of MOLT-4 cells, we could not detect any enhancement of syncytia formation, over that seen with wild-type LAI particles. This result should alleviate concerns of patients on highly active antiretroviral therapy (HAART), that protease inhibitors might accelerate progression of the disease through enhanced production of defective, 'immature'-appearing particles. (C) 2000 Elsevier Science B.V. All rights reserved.ELSEVIER SCIENCE BV, 2000年02月, VIRUS RESEARCH, 66(2) (2), 131 - 137, 英語[査読有り]研究論文（学術雑誌）
- Exposure of normal monocyte-derived dendritic cells to human immunodeficiency virus type-1 particles leads to the induction of apoptosis in co-cultured CD4(+) as well as CD8(+) T cellsThe depletion of immune T cells by human immunodeficiency virus type-1 (HIV-1) infection is a major mechanism involved in the pathogenesis of AIDS. Here, we examined a possible effector function of blood monocyte-derived dendritic cells (DCs) to induce apoptosis in bystander CD4(+) and CD8(+) T cells. The DCs were generated by culturing monocytes in the presence of granulocyte-macrophage colony-stimulating factor and interleukin-4, The DCs exposed to HIV-1 particles were co-cultured with healthy donor-derived blood T cells at a ratio of 1:20, Analyses by percent cell mortality, staining with propidium iodide and reactivity with Annexin V revealed the induction of apoptosis in both CD4(+) and CD8(+) target T cells. Further, this apoptosis occurred without stimulation with mitogens when the cell cycle of target T cells shifted from G(0) to G(1), probably due to the mitogenic effect of the DCs, Thus, induction of apoptosis in both CD4(+) and CD8(+) T cells occurred via interaction with DCs adsorbed with HIV-1 particles.CENTER ACADEMIC PUBL JAPAN, 2000年, MICROBIOLOGY AND IMMUNOLOGY, 44(2) (2), 111 - 121, 英語[査読有り]研究論文（学術雑誌）
- Phorbol 12-myristate 13-acetate (PMA)-induced HIV-1 production in U1 cells was markedly suppressed by inhibitors of poly (ADP-ribose) polymerase (PARP), Northern blot analysis revealed that the PARP-inhibitors suppressed the virus production at a level of transcription, In order to examine the effect of PARP on transcriptional regulation of HIV-1 genes, we transfected a reporter plasmid containing HIV-1-LTR-promoted luciferase gene to L-1210 cell clones, which expressed varying decreased level of PARP, In wild type L-1210 cells, the expression of LTR-promoted luciferase gene was stimulated approximately 4-fold in response to PMA, whereas the PMA-dependent response was almost abolished in mutant cells, which expressed only 8% of PARP of the wild type cells. The effect of decrease in PARP content on the function of HIV-1-LTR was confirmed also in human wild type cells, Jurkat and J111, which were co-transfected with the reporter plasmid and a plasmid expressing a PARP-antisense RNA: Down-regulation of PARP in the cells by the expression of the antisense RNA significantly suppressed the PMA-dependent, LTR-function of the reporter plasmid in both Jurkat and J111 cells. NF-kappa B, which is known to mediate the PMA-induced activation of HIV-1 in U1 cells, was found to be activated approximately 5-fold in PMA-treated U1 cells. PARP-inhibitor, unexpectedly, did not suppress but rather stimulated (approximately 2-fold) the NF-kappa B activation. Combining the results with the finding that the LTR function was minimum in a PARP-defective mutant cells in spite of a very high level of the activated NF-kappa B in the cells, we suggest that PARP, in addition to activated NF-kappa B, is essential for the function of HIV-1 LTR. (C) 1999 Academic Press.ACADEMIC PRESS INC, 1999年08月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 262(1) (1), 285 - 289, 英語[査読有り]研究論文（学術雑誌）
- Purified recombinant HIV-1 Tat protein stimulated acceptor-dependent reaction of poly(ADP-ribose) polymerase in a dose-dependent manner. Analysis of the reaction products by SDS-polyacrylamide gel electrophoresis followed by immunoblotting with anti-poly(ADP-ribose) antibody revealed that recombinant Tat proteins were covalently modified with poly(ADP-ribose) in the enzyme reaction. Even though no significant effect of the modification was detected in the activity of Tat to form a specific complex with TAR (a viral transactivation response element) RNA, the present results raise the possibility that poly(ADP-ribose) polymerase is involved in the regulation of HIV-1 through the modification of a virus-encoded transactivator, Tat protein. (C) 1999 Academic Press.ACADEMIC PRESS INC, 1999年07月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 261(1) (1), 90 - 94, 英語[査読有り]研究論文（学術雑誌）
- Treatment of 26L cells, a subclone obtained from U937 cells, with TNF-alpha or DNA-damaging agents such as teniposide (VM26) and camptothecin (CPT) induced morphologically and biochemically typical apoptotic changes, including the activation of procaspase-3. The cells persistently infected with HIV-1 (26L/HIV), however, showed a marked resistance to VM26 and CPT, whereas they hardly lost the sensitivity to TNF-alpha. TNF-alpha-induced apoptosis of 26L/HIV cells proceeded without the increase in caspase-3 activity, indicating that signaling for apoptosis in the infected cells proceeded through an alternative caspase-3-independent pathway which could respond to TNF-alpha but not to VM26 and CPT. The evidence that p-toluenesulfonyl-L-lysine chloromethyl ketone (a trypsin-like serine protease inhibitor) blocked VM26- and CPT-induced apoptotic changes but not TNF-alpha-induced apoptosis also supported the existence of the alternative TNF-alpha-inducible pathway. The results also suggest that a TLCK-sensitive protease is involved upstream of the procaspase-3 activation process and that the protease is essential for the progress of VM26- and CPT-induced apoptosis. The similar effect of HIV-1-productive infection on the apoptosis induced by the DNA-damaging agents was also confirmed by utilizing U1 cells, which are latently HIV-1-infected U937 cells. The cells became resistant to these agents after induction of the viral production by pretreatment with PMA. These results suggest that persistent HIV-1 infection blocks an apoptotic pathway triggered by DNA damaging agents through the inhibition of the procaspase-3 activation process. (C) 1999 Academic Press.ACADEMIC PRESS INC, 1999年03月, EXPERIMENTAL CELL RESEARCH, 247(2) (2), 514 - 524, 英語[査読有り]研究論文（学術雑誌）
- 奈良医学会, 1998年12月, Journal of Nara Medical Association, 49(6) (6), 495 - 495, 日本語PARPがHIV複製に及ぼす影響
- Apoptosis is an important mechanism of human immunodeficiency virus type 1 (HIV-1)-induced T-cell depletion. Our recent findings revealed mitogenic stimulation-dependent apoptosis induction in healthy donor-derived peripheral blood T-lymphocytes after adsorption with defective HIV-1 particles through acquirement by a subset of CD4(+)/CD38(-) cells of specific killer function. Based on these in vitro observations, we have extended the significance of this killing activity of CD4(+)/CD38(-) cells directly derived from HIV-1 carriers. The CD4(+)/CD38(-) cells from HIV-1-positive individuals showed significantly higher cell-killing activities than those from HIV-1-negative donors by co-culture with allogeneic resting T-cells after mitogenic stimulation. Furthermore, most of the samples induced apoptosis in a Fas-dependent manner. Thus, it is suggested that HIV-1 infection-related apoptosis is triggered by inappropriate activation of a certain resting T-cell subset, presumably due to adsorption with HIV-1 particles. (C) 1998 Elsevier Science B.V. All rights reserved.ELSEVIER SCIENCE BV, 1998年07月, VIRUS RESEARCH, 56(1) (1), 115 - 122, 英語[査読有り]研究論文（学術雑誌）
- Dependence on host cell cycle for activation of human immunodeficiency virus type 1 gene expression from latencyHuman immunodeficiency virus type 1 (HIV-1) establishes latent infection of a certain population of CD4(+) host cells which could be long-term reservoirs for HIV-1, The expression of viral genes in such long-term infected cells is strongly regulated by cellular status, such as the phase of the cell cycle or stage of cell differentiation. Here, viral gene expression in synchronized U1 cells, a monocytic cell clone latently infected with HIV-1, was characterized. The expression of HIV-1 antigens was detected exclusively at G(2)/M phase in U1 cells, irrespective of phorbol myristate acetate (PMA) treatment. The induction of HIV-1 gene expression in PMA-treated cells was due to the recruitment of NF-kappa B with DNA-binding activity at G(2)/M phase. Activated NF-kappa B was induced only by PMA treatment during the late G(1) to S, but not after entering G(2) phase, indicating that the transcriptional factor(s) involved in viral gene expression is also largely regulated by the host cell cycle, In contrast, the enhancement of antigen expression by treatment with tumour necrosis factor-alpha (TNF-alpha) was cell cycle-independent. In fact, NF-kappa B was activated 2 h after TNF-alpha treatment at all stages of the cell cycle, Thus, the mechanisms of HIV-1 activation from latency in U1 cells by PMA and TNF-alpha treatment are different. The model system using U1 cells shown here may provide insight into the mechanisms responsible for HIV-1 gene expression from latency.SOC GENERAL MICROBIOLOGY, 1998年06月, JOURNAL OF GENERAL VIROLOGY, 79, 1363 - 1371, 英語[査読有り]研究論文（学術雑誌）
- Human immunodeficiency virus type 1 (HIV-1) infection indirectly induces activation-dependent apoptosis in bystander immune CD4(+) T-cells, a hallmark of AIDS pathogenesis. It is well known that this pathogenetic event is significantly correlated with a high virus load. Active viral replication occurs in HIV-1 asymptomatic carriers throughout all stages of clinical disease. Most of the HIV-1 in plasma is derived from short-lived infected cells with a half life of a few days; however, a minor population of virus is derived from long-lived persistently and latently infected cells. Recently, the importance of such latent reservoirs for HIV-1 has come to the forefront because of studies with potent antiretroviral inhibitors that block only new rounds of infection. An initial large drop in viral load occurs within two weeks as noted by a decrease in plasma viremia. This is then followed by a slower second-phase decay, since only a small fraction of latently infected resting CD4(+) T-cells carry replication-competent, integrated provirus. This review highlights the mechanisms of apoptosis induction in bystander immune cells by both protease-defective, gp120-containing HIV-1 particles, as well as by wild-type virus that appears to be derived predominantly from long-lived infected cells. A model involving the NH2-terminal Nef domain (p7) in this 'bystander apoptosis' event is also presented. (C) 1997 Elsevier Science B.V.ELSEVIER SCIENCE BV, 1997年12月, VIRUS RESEARCH, 52(2) (2), 145 - 156, 英語[査読有り]
- A significant level of superoxide (O-2(-)) generation was observed in a U937-derived subclone following infection with human cytomegalovirus (HCMV). Although there were no detectable levels of viral mRNA and/or protein expression, HCMV DNA content transiently increased immediately before O-2(-) generation. Similarly, O-2(-) generation was also observed in peripheral blood monocytes derived from healthy donors. (C) 1997 Elsevier Science B.V.ELSEVIER SCIENCE BV, 1997年10月, IMMUNOPHARMACOLOGY, 37(2-3) (2-3), 185 - 190, 英語[査読有り]研究論文（学術雑誌）
- Apoptosis is a major mechanism whereby HIV-1 depletes uninfected CD4(+) and CD8(+) T cells, We previously showed that resting peripheral blood T cells derived from healthy donors were killed by an apoptotic mechanism after adsorption to gp120-containing, protease-defective HIV-1 (L-2) particles, more effectively than parental wild-type LAI adsorption or rgp120-mediated CD4 crosslinking, followed by mitogenic stimulation, Here, we present evidence that the L-2 particle-based apoptosis was induced both in CD4(+) and CD8(+) cells by generation of effector cells which were mainly derived from a resting memory CD4(+)CD38(-) subset, This subset enhanced the CD25 expression on the surface and secreted IFN-gamma in the culture supernatant after L-2 particle exposure. Significant elevation of Fas ligand mRNA was found in the subset by L-2 particle exposure, while expression of Fas antigen on uninfected T cells was induced by exposure to IFN-gamma. These results indicate that L-2 particles can shift the CD4(+)CD38(-) subpopulation from a resting to an activated state, and this activation leads to killing of bystander CD4(+) and CD8(+) T cells by a Fas-mediated mechanism, In fact, purified CD4(+)CD38(-) cells exposed to L-2 particles were converted into effector cells that were able to kill autologous as well as allogenic target T cells pretreated with IFN-gamma. Further, we found that the observation of apoptosis due to L-2 particles was a more general phenomenon, that also occurred with Thai primary HIV-1 isolates, These results suggest that such specific types of HIV-1 particles may play a major role in the induction of apoptosis for both bystander CD4(+) and CD8(+) T cells, through inappropriate activation of CD4(+)CD38(-) cells.OXFORD UNIV PRESS, 1997年10月, INTERNATIONAL IMMUNOLOGY, 9(10) (10), 1453 - 1462, 英語[査読有り]研究論文（学術雑誌）
- A polyclonal antibody against purified bull seminal plasma Ca2+, Mg2+-dependent endonuclease was raised in a rabbit. The antibody specifically cross-reacted with chromatin-bound Ca2+, Mg2+-dependent endonucleases from bovine thymus, human placenta, and bovine, rat and mouse liver in addition to the bovine seminal enzyme. The antibody did not cross-react with other endonucleases examined, including the acid-endonucleases from bovine thymus and liver, porcine spleen DNase II, micrococcal nuclease, and bovine pancreas DNase I, a known Ca2+ and Mg2+ requiring endonuclease. The present results indicate that this antibody specifically recognizes a class of so-called Ca2+, Mg2+-dependent endonuclease, which is localized in cell nuclei of various tissues and is probably involved in chromatin degradation during apoptosis, The antibody will be used to study the functional role of this class of endonuclease. (C) 1997 Academic Press.ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS, 1997年07月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 236(2) (2), 423 - 426, 英語[査読有り]研究論文（学術雑誌）
- Cell-mediated immune responses constitute a major defense against the spread of human immunodeficiency virus type I (HIV-I). However, multiple alterations within a particular epitope may accumulate during disease progression, allowing the virus to escape cytotoxic T lymphocytes (CTLs). Therefore, the best candidate for a peptide vaccine that would prevent the onset of the disease might be a chain section containing epitopes for the generation of CTLs in regions of conserved sequences among different HIV-I isolates. We previously showed that immunizing mice with synthetic peptides consisting of 23-amino acids (Gag-23mer; 287-309 amino acid residues) in a highly conserved region derived from the major core protein p24 of HIV-I generates specific CTLs as well as antibodies. Here, we identified one CTL (T-1; 291-300) and two B-cell (B-1; 290-299 and B-2, 300-309) epitopes, all of which consisted of 10 amino acids within the region. In addition, helper T cells primed by the Gag-23mer peptide were proliferated by in vitro stimulation with a 21mer (H-1, 289-309) or a 19mer (H-2; 291-309) peptide, but not with a 17mer peptide (293-309) or 19mer peptide (287-305). Immunization with the H-I peptide generated an antibody reactive to B-I, but not B-2, whereas that with H-2 generated an antibody reactive to B-2, but not B-1. CTLs were not generated by immunization with these peptides, indicating that the entire sequence of Gag-23mer is the helper epitope for CTLs. Thus, the Gag-23mer is a chain section containing epitopes for cytotoxic T, B and helper T-cells within a highly conserved region of HIV-I Gag protein. (C) 1997 Elsevier Science Ltd.ELSEVIER SCI LTD, 1997年04月, VACCINE, 15(5) (5), 489 - 496, 英語[査読有り]研究論文（学術雑誌）
- MARY ANN LIEBERT INC PUBL, 1997年04月, AIDS RESEARCH AND HUMAN RETROVIRUSES, 13(6) (6), 523 - 526, 英語[査読有り]研究論文（学術雑誌）
- Insertions, duplications and substitutions in restricted gp90 regions of equine infectious anaemia virus during febrile episodes in an experimentally infected horseWe have studied a horse which exhibited typical clinical signs of disease when experimentally infected with a non-adapted virulent strain of equine infectious anaemia virus (EIAV), designated V70. Five viruses (F1V, F2V, F3V, F4V and F5V) were recovered during periodic febrile episodes, Cross-neutralization tests revealed that all of these variants and the parental V70 were antigenically distinct, Sequencing of their full-length env gp90 genes and gp45 5' sequences revealed novel mutations at a limited number of nucleotide positions, consisting of insertions and duplications in the gp90 principal neutralizing domain (PND) in F1V, F3V and F5V, Parts or all of small units (6, 9 and 12 nucleotides) located just before the insertion site were used for the duplications, Furthermore, amino acid substitutions in the env PND and hypervariable region were also observed in all five viruses, These mutations may contribute to the generation of serial variants. Consequently, the full-length gp90 sequences showed close relationships between V70, F2V and F4V, and between F1V, F3V and F5V. In addition to the two domains (PND and hypervariable region), a comparison of these viruses with the reported env gp90 sequences revealed four additional variable domains, although these four domains were highly conserved among the five variants.SOC GENERAL MICROBIOLOGY, 1997年04月, JOURNAL OF GENERAL VIROLOGY, 78, 807 - 820, 英語[査読有り]研究論文（学術雑誌）
- In asymptomatic carriers, the vast majority of human immunodeficiency, virus type I (HIV-IJ infection is non-productive whilst the clinical stage of disease is associated with significant virus expression. Virus-specific CD8(+) T-cell functions are believed to play a major role in the generation of heterogeneous virus populations and in subsequent disease progression. Here, we prepared two types of HIV-1 isolate by culturing whole and CD8(+) T cell-depleted peripheral blood mononuclear cells (PBMCs) from five asymptomatic carriers. The former is expected to be escape variant populations, whereas the latter would be mixed populations including the former viruses. The analyses of Nef and Env V3 sequence variations of viruses in a total of 77 and 44 DNA clones, respectively, allowed a direct comparison to be made of the differences between the paired isolates. Comparison of Nef sequences between the paired isolates showed them to be more distinct in two carriers with a relatively stable CD4/CD8 ratio (Nos 68 and 69), than in two other carriers with similar CD4/CD8 ratios (Nos 53 and 57), or in carrier No. 67, which had an extremely lower CD4/CD8 ratio. By contrast, a distinction between the paired isolates by use of the Env V3 sequences was only apparent in the latter three carriers. These results indicate that the predominant populations of HIV-1 in Nos 68 and 69 were sensitive to selective pressure from Nef-specific CD8(+) T-cells, while those in Nos 53, 57, and 67 were sensitive to pressure from V3-specific CD8(+) T-cells. It is noteworthy that Nos 53 and 57 progressed to an AIDS-related complex shortly and several years after this examination. These data suggest that HIV-1-induced pathogenesis is more strongly associated with the generation of variant nef alleles than with env V3 variants, and that these arise by CD8(+) T-cell pressure. (C) 1997 Elsevier Science Ltd.ELSEVIER SCI LTD, 1997年04月, VACCINE, 15(5) (5), 497 - 510, 英語[査読有り]研究論文（学術雑誌）
- Protease-defective, gp120-containing human immunodeficiency virus type 1 particles induce apoptosis more efficiently than does wild-type virus or recombinant gp120 protein in healthy donor-derived peripheral blood T cellsApoptosis and syncytium formation are two mechanisms by which human immunodeficiency virus type 1 (HIV-1) impairs uninfected CD4(+) T-cell function and are mainly involved in the progression of the disease to AIDS, Previously, we showed that gp120-containing, protease-deficient HIV-1 (L-2) particles generated syncytia by particle-mediated fusion with uninfected cultured CD4(+) T cells, Here, we present evidence that such L-2 particles can induce apoptosis in 40 to 50% of T cells which were enriched from HIV-1-negative healthy donor-derived peripheral blood mononuclear cells (PBMC-Ts), Activation of PBMC-Ts with phytohemagglutinin, concanavalin A, or ionomycin after incubation with L-2 particles resulted in the loss of proliferative capacity and gradual induction of apoptosis over 3 days, Wild-type strain LAI particles or recombinant gp120 were markedly less efficient (less than or equal to 15%) at inducing such apoptosis, Western blot (immunoblot) analysis revealed that L-2 particles contained a larger amount of Env gp120 than LAI particles, Either preincubation of PBMC-Ts with a Fas antagonist or preincubation of L-2 particles with soluble CD4 blocked most of the apoptosis. This suggests that L-2-like particles can play a major role in HIV-1-induced apoptosis of uninfected bystander cells.AMER SOC MICROBIOLOGY, 1997年01月, JOURNAL OF CLINICAL MICROBIOLOGY, 35(1) (1), 41 - 47, 英語[査読有り]研究論文（学術雑誌）
- Intracellular glutathione as a possible direct blocker of HIV type 1 reverse transcriptionIn AIDS patients, chronic inflammation and elevated levels of cytokines seem to be associated with reduced levels of glutathione (GSH). GSH has been proposed to inhibit the activation of NF-kappa B, which results in the inhibition of HIV-1 replication. Here, we show the evidence that GSH and N-acetylcysteine, but not L-cysteine or dithiothreitol, could inhibit the reverse transcriptase (RT) process of HIV-1. Such inhibition was not observed with the RT of murine leukemia virus.MARY ANN LIEBERT INC PUBL, 1996年11月, AIDS RESEARCH AND HUMAN RETROVIRUSES, 12(17) (17), 1635 - 1638, 英語[査読有り]研究論文（学術雑誌）
- Several recent reports support the hypothesis that apoptosis occurring in leukocytes of human immunodeficiency virus type 1 (HIV-1)-infected individuals is important in progression to AIDS. Specifically, apoptosis of uninfected bystander cells appears critical in the pathogenesis of disease. Here, we present evidence that protease-defective, gp120-containing HIV-1 (L-2) particle preparations specifically induce apoptosis in cells obtained from a subset of promonocytic U937-derived subclones, The rate of apoptosis induction was inversely correlated with the susceptibility of the U937 subclones to wild-type HIV-1 infection. Three types of apoptosis experiments were performed: DNA content analysis by flow cytometry, apoptotic nuclear degradation by fluorescent microscopy and DNA fragmentation analysis by agarose gel electrophoresis. Kinetic analysis revealed that there was a slower induction of apoptosis by L-2 particle preparations than with tumor necrosis factor (TNF)-alpha or anti-fas antibody. However, there were no significant differences in the initial binding rates of L-2 particles as well as the binding of TNF-alpha or anti-fas antibody to the U937 subclones. The basal level of protein kinase C activity was higher in high-type subclones compared with low-type subclones. These results suggest that U937 cells can be divided into at least two subpopulations, one that permits a productive HIV-1 infection but is not subjected to L-2 particle preparation-induced apoptosis, while the other poorly replicates HIV-1 and is subjected to L-2 mediated apoptosis, although at a slower rate than found with TNF-alpha or anti-fas antibody.OXFORD UNIV PRESS UNITED KINGDOM, 1996年11月, INTERNATIONAL IMMUNOLOGY, 8(11) (11), 1687 - 1697, 英語[査読有り]研究論文（学術雑誌）
- 1996年07月, [Hokkaido igaku zasshi] The Hokkaido journal of medical science, 71(4) (4), 489 - 508, 日本語[Studies on susceptibility of promonocytic cell line U937-derived subclones to HIV-1 infection and apoptosis induction].[査読有り]
- The recombinant bacillus Calmette-Guerin (rBCG) secretion system utilizing an extracellular alpha antigen of Mycobacterium kansasii (alpha-K) was characterized biochemically and immunologically. The human immunodeficiency virus type1 (HIV-1)p17(gag) B cell epitope fused to alpha-K was secreted in extremely large amounts. At least three mice out of seven inoculated with rBCG generated high titres of antibody to the epitope. The long-lasting antibody production persisted more than 14 months.BLACKWELL SCIENCE LTD, 1996年02月, Scandinavian journal of immunology, 43(2) (2), 202 - 209, 英語[査読有り]研究論文（学術雑誌）
- High susceptibility of U937-derived subclones to human immunodeficiency virus type 1 infection correlates with accumulation of unintegrated circular viral DNAOur previous report showed that U937-derived subclones were differentiated into at least three types (high, middle, and low types), even in the subclones expressing similar levels of surface CD4, in terms of the kinetics of the appearance of viral antigens and virus production after infection with human immunodeficiency virus type 1 (HIV-I). Here we showed the evidence that high susceptibility to HIV-1 infection, which was confirmed by the profound expression of viral messages and antigens, was exclusively associated with a high number of the unintegrated extrachromosomal form of viral DNA, but not with the amounts of adsorbed virus RNA nor those of integrated DNA form. The difference in the amounts of extrachromosomal form of viral DNA was also observed in the culture with 3'-azido-3'-deoxythymidine (AZT), indicating that the susceptibility is essentially unrelated to multiple infection events. Thus, the susceptibility of U937-derived subclones to HIV-1 infection seems to be affected by the occurrence of specific events involved in the accumulation of unintegrated viral DNA after viral adsorption.KLUWER ACADEMIC PUBL, 1996年, VIRUS GENES, 12(2) (2), 117 - 129, 英語[査読有り]研究論文（学術雑誌）
- EXTRACELLULAR NEF PROTEIN REGULATES PRODUCTIVE HIV-1 INFECTION FROM LATENCYIn HIV-1-infected asymptomatic carriers, the vast majority of infected cells in PBMCs are believed to be latently or nonproductively infected. We have isolated a subclone (MOLT-20-2) from an infected T cell line that expressed HIV-1 Ags at a very low level. However, viral Ag expression was markedly up-regulated by stimulation with either TNF-alpha A23187, or PMA, indicating that the subclone might provide a suitable model of HIV-1 latency. Our previous studies have shown that the carboxyl-terminal region of the extracellular form of HIV-1 Nef played an important role in the interaction of infected cells with uninfected T cells, and could induce the cytostatic state. This suggested that Nef might contribute to intracellular signal transduction through an interaction with latently infected cells. We show in this study that stimulation of MOLT-20-2 with soluble Nef leads to HIV-1 activation from latency in a dose-dependent manner. Moreover, using a total of 14 overlapping Nef-related synthetic peptides, stimulatory activity was mapped to a discrete peptide (amino acid residues 132-147) that had the potential to activate latent HIV-1. This novel Nef function was confirmed by activation of virus production from the PBMCs of asymptomatic carriers. In addition, Nef-dependent HIV-1 activation from latency was also observed in another independently derived, latently infected cell line, U1, though not in cell line ACH-2. These results extend the significance of the Nef activity in vivo to the regulation of productive HIV-1 infection from latency, and define the regions of the protein involved.AMER ASSOC IMMUNOLOGISTS, 1995年12月, JOURNAL OF IMMUNOLOGY, 155(11) (11), 5289 - 5298, 英語[査読有り]研究論文（学術雑誌）
- We previously showed that superoxide (O-2(-)) significantly enhanced human immunodeficiency virus type 1 (HIV-l)-induced syncytia formation in co-cultured infected and uninfected human T cells. In this study, we describe a novel chemotactic response of uninfected CD4(+) T cells by stimulating infected T cells with O2(-). Syncytia formation was amplified only when persistently infected cells were stimulated by O-2(-). When the infected cells in lower well of microplate were cultured with uninfected cells in the upper well of a Boyden chamber with 8.0 mu m pores, uninfected cell migration to the porous membrane was significantly amplified by stimulating infected cells with O-2(-). In contrast, similar functions were slight under the same assay conditions in the presence of known chemokines such as human RANTES and macrophage inflammatory protein 1 (MIP-1 alpha and beta), which all activate T lymphocytes. In addition, it is unlikely that the O-2(-)-induced chemotactic response is due to soluble HIV-1 proteins from infected cells or to amplified expression levels of cell surface functional molecules such as CD4 and LFA-1 (CD11a and CD18) as well as HIV-1 Env gp120 on uninfected and/or infected cells. Thus, an unknown chemotactic factor could be generated from infected T cells by stimulation with 0(2)(-) and it might contribute to viral transmission by activating cell-to-cell interactions.ELSEVIER SCIENCE BV, 1995年11月, IMMUNOPHARMACOLOGY, 31(1) (1), 73 - 84, 英語[査読有り]研究論文（学術雑誌）
- INHIBITION AND DOWN-REGULATION OF POLY(ADP-RIBOSE) POLYMERASE RESULTS IN A MARKED RESISTANCE OF HL-60 CELLS TO VARIOUS APOPTOSIS-INDUCERSIn a previous report we described that adenosine-induced apoptosis of HL-60 cells was blocked by the pretreatment of cells with a potent inhibitor (3-aminobenzamide) of poly(ADP-ribose) polymerase (PARP). The pretreatment of the cells with nicotinamide, another inhibitior of the enzyme, also suppressed most effectively the adenosine-induced apoptosis. This inhibition was reversible and observed during apoptosis mediated by other known apoptosis inducers such as actinomycin D and staurosporine (group I inducers), but nicotinamide was ineffective on the apoptosis mediated by VM 26, camptothecin and A23187 (group Ii inhibitors). In addition to the enzyme inhibition, a down-regulation of the enzyme level caused by the pretreatments of cells with differentiation-inducing agents, retinoic acid (RA) and dimethylsulfoxide (DMSO) also resulted in a marked resistance of the cells to the apoptosis inducers. A pretreatment of the cells for a limited time of 24 hrs. by these agents decreased the PARP level to 66-75% of the untreated cells and the cells showed a quite similar resistance to the group I apoptosis inducers like the cells treated with the enzyme inhibitors, whereas they were still sensitive to the group II inhibitors. A more prolonged treatment for 48 hrs. of the cells with RA and DMSO resulted in further down-regulation of the cellular PARP reaching respectively 50 and 43% of control cells and at this stage, the cells became resistant to all the inducers of both groups. These results suggest that the pathway, by which both groups of the inducers initiate and progress apoptosis, is not identical but include at least two different processes which are differently affected by PARP-inhibition or by different levels of cellular PARP.CELLULAR & MOLECULAR BIOLOGY, 1995年09月, CELLULAR AND MOLECULAR BIOLOGY, 41(6) (6), 771 - 781, 英語[査読有り]研究論文（学術雑誌）
- Poly(ADP-ribose) polymerase, a nuclear enzyme, is suggested to be involved in apoptotic cell death. It is also known that apoptotic cell death following HIV-1 infection is the most important feature of AIDS pathogenesis. Thus, to evaluate the relations between the enzyme and HIV-1 infection, we examined the enzyme activity of several subclones of human promonocytic cell line U937, which showed different susceptibility to HIV-1 infection. The nuclear extracts of two ''high type clones'' (possessing high susceptibility to HIV-1 infection) contained approximately 4 to 7-fold less enzyme than two low type clones when assayed under a full activation of enzyme. Parent clone, possessing an intermediate susceptibility to HIV-1, showed an intermediate enzyme level, suggesting that low level of this enzyme in cells is important for an effective infection of HIV-1. Furthermore, when these U937 subclones persistently infected with HIV-1 were examined, a dramatic decrease of the enzyme activity, reaching 2 to 16% of uninfected cells, was observed in all of these clones. The levels of poly(ADP-ribose) glycohydrolase in these clones were relativity unchanged. Activity gel analysis and immunoblotting of the enzyme in the clones revealed that the low enzyme activities observed in uninfected ''high type clones'' and all HIV-1-infected clones were due to a marked decrease of the enzyme protein itself. All of these results suggest that HIV-1 infection involves some mechanism to downregulate cellular poly(ADP-ribose) polymerase and that a lower level of the enzyme may be essential for an effective production of the virus and/or for a stable virus/host interaction. (C) 1995 Academic Inc, Press.ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS, 1995年08月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 213(1) (1), 161 - 168, 英語[査読有り]研究論文（学術雑誌）
- The promonocytic human leukemic cell line U937, when infected with lymphotropic human immunodeficiency virus type 1 (HIV-1), becomes a continuous virus producer. A total of 46 U937-derived subclones in suspension was isolated and classified into three (2 high, 42 middle, and 2 low) types based on their susceptibility to the infection. By analyzing subclones before infection, we found that the high-type subclones expressed LFA-1 antigens at a relatively low level. In addition, the ability of these subclones to induce adherence after exposure to phorbol 12-myristate 13-acetate (PMA) was reduced. In contrast, a transition by HIV-1 infection to adherent macrophage-like cells was induced only in the high-type, but not in the low-type subclones. The high-type adherent cells obtained by HIV-1 infection were followed by further lineage to become retrodifferentiated suspension cells showing reduced syncytia formation ability. Superoxide was generated in the high-type subclones, without PMA-mediated differentiation, from the early stage of infection before HIV-1 replication, as well as during undifferentiated, differentiated and retrodifferentiated stages. In contrast, it was only transiently generated at acute phase of HIV-1 replication in low-type subclones. Long-term culture of the low-type subclones decreased the expression of major structural viral protein Gag and also virus production. Thus, the mechanism by which PMA differentiates U937 cells is not the same as that induced by HIV-1 infection. The latter mechanism results in high susceptibility to infection. The HIV-1 phenotypes of finally obtained persistently infected cells were also affected by the cell stages at the time of infection.ELSEVIER SCIENCE BV, 1995年06月, IMMUNOPHARMACOLOGY, 30(1) (1), 89 - 101, 英語[査読有り]研究論文（学術雑誌）
- To determine the mechanism underlying the human immunodeficiency virus type 1 (HIV-1) latency and its activation in monocyte/macrophage lineage, the human promonocytic cell line U937 was infected with HIV-1 after differentiation with varied doses of phorbol 12-myristate 13-acetate (PMA). Variously differentiated intermediate stages were generated in U937 cells in a dose-dependent manner. When these cells were infected with lymphotropic HIV-1, the kinetics of the production of HIV-1 DNA, the appearance of HIV-1 antigen-positive cells, and viral production in the conditioned media were slower at higher doses of PMA. This different susceptibility to the infection was not due to the rate of HIV-1 adsorption. Viral replication from latency in the differentiated cells was activated in proportion with the retrodifferentiation observed in long-term cultures of the host cells. Thus, our data demonstrate the close correlation between the regulation of HIV-1 replication and the differentiation stage of monocyte/macrophage lineage cells at the time of HIV-1 infection. The retrodifferentiation phenomenon in infected cells seems to be particularly important for understanding the mechanisms for HIV-1 activation from latency.ELSEVIER SCIENCE BV, 1995年06月, IMMUNOPHARMACOLOGY, 30(1) (1), 27 - 39, 英語[査読有り]研究論文（学術雑誌）
- THE CARBOXYL-TERMINAL REGION OF HIV-1 NEF PROTEIN IS A CELL-SURFACE DOMAIN THAT CAN INTERACT WITH CD4(+) T-CELLSOur previous studies have shown that the HIV-1 Nef Ag is expressed, at least in part, on the surface of infected cells. We demonstrated this by using membrane immunofluorescence and flow cytometry with Nef murine mAbs. To identify the domain of Nef exposed on the cell surface, epitope mapping of these and a new mAb was performed by ELISAs by using several recombinant truncated Nef fusion proteins and synthetic peptides. The results showed that mAbs F1, E7, E9, and 4H4 recognized Nef epitopes located at amino acid residues 148-157, 192-206, 158-206, and 1-33, respectively. The intensity of cell surface Nef staining was stronger with mAbs E7 and E9 than with F1, and there was no staining by 4H4, which indicates that the carboxyl-terminal region of Nef is predominantly exposed on the surface of HIV-1-infected T cell lines and PBMC. This surface Nef domain displayed high affinity for the surface of uninfected CD4(+) T cells; because the binding of a soluble form of recombinant Nef protein to the cell surface was specifically blocked by the E7 and E9 mAbs or by synthetic peptides that contained the carboxyl-terminal region of Nef. In addition, syncytium formation between infected and uninfected cells also was specifically reduced by the same mAbs or peptides. Thus, the cell surface domain of Nef seems to play an important role in the interaction between HIV-1-infected and CD4(+) uninfected T cells.AMER ASSOC IMMUNOLOGISTS, 1994年12月, JOURNAL OF IMMUNOLOGY, 153(12) (12), 5826 - 5837, 英語[査読有り]研究論文（学術雑誌）
- Cell-mediated immune responses are a major immune defence mechanism against the spread of human immunodeficiency, virus type 1 (HIV-1) which may lead to acquired immune deficiency syndrome (AIDS). Therefore, the best candidate for a peptide vaccine preventive from the onset of the disease might be a chain section containing both B- and T-cell epitopes in regions of conserved sequences between the different HIV-1 isolates. We previously identified the highly conserved linear B-cell epitope (23 amino acids in the major core protein p24). Since the epitopes of cytotoxic T lymphocytes (CTLs) can be defined by short synthetic peptides, we examined whether this highly, conserved region can elicit viral-specific, cell-mediated immune responses. The results showed specific induction of CD8 + CTLs in mice by immunization with the Gag 23-mer peptide. Lysis of targets is specific since unpulsed cells with the same MHC haplotype or cells with a different MHC haplotype pulsed with the peptide were resistant to lysis. This in vivo response induced by the Gag 23-mer peptide was almost the same as that induced by the 15-amino acid peptide from the HIV-1 Env gp120 which is an immunodominant domain in the V3 loop. Lymphocyte proliferation of T-cell fraction from immune spleen cells was observed after in vitro stimulation with the Gag 23-mer peptide, whereas there was no apparent lymphocyte proliferation with the Env 15-mer peptide. In addition, specific antibodies were raised against Gag p24 in mice immunized with the Gag 23-mer peptide. Thus, it was suggested that the Gag 23-mer peptide contained both helper and killer epitopes as well as the binding region to murine class I-MHC, H-2(d). In addition, guinea-pigs primed with the same Gag 23-mer peptide showed stronger delayed-type hypersensitivity (DTH) than with the Env 15-mer peptide. These results demonstrate the applicability of this Gag 23-mer peptide as the prognostic indicator antigen for immunoreactivity by DTH and as a peptide vaccine which might prevent the onset of AIDS in HIV-1 carriers.BUTTERWORTH-HEINEMANN LTD, 1994年05月, VACCINE, 12(6) (6), 485 - 491, 英語[査読有り]研究論文（学術雑誌）
- The host immune response of cell-mediated immunity, particularly that of cytotoxic T lymphocytes (CTLs), is a major immune defence mechanism which may provide resistance to a human immunodeficiency virus type 1 (HIV-1) spread leading to acquired immune deficiency syndrome (AIDS). To prevent the accompanying activity of HIV-1 proteins responsible for the loss of helper T-lymphocyte function, it is crucial to develop a live attenuated recombinant vaccine expressing only T- or both T- and B-cell epitopes. Here, we examined the expression of the HIV-1 Env protein V3 region (15 amino acids from Arg(315) to Lys(329)) in Mycobacterium bovis BCG as a fused form with an extracellular alpha antigen of Mycobacterium kansasii. Balb/c mice inoculated with this recombinant BCG (rBCG), rapidly induced V3 peptide-specific CTLs. Target cell lysis was restricted to the murine class I major histocompatibility complex, H-2(d). A similar CTL response was also elicited after Balb/c mice were immunized with the same rBCG even when pre-inoculated with non-recombinant BCG. Thus, the rapid induction of HIV-1-specific CTLs indicates that this vaccine may be a therapeutic approach to preventing progression to AIDS.BUTTERWORTH-HEINEMANN LTD, 1994年02月, Vaccine, 12(2) (2), 153 - 158, 英語[査読有り]研究論文（学術雑誌）
- Oxidative stress is thought to be involved in the progression of human immunodeficiency virus type 1 (HIV-1)-induced disease. We examined the effect of superoxide (O2-) on HIV-1 spread in cultured human CD4+ cell lines. The O2- significantly enhanced cell-to-cell transmission of HIV-1, although its effect on HIV-1 replication was not evident, presumably due to its cytostatic activity. The effect was notable on the HIV-1 transmission from macrophages to T lymphocytes by endogenous, macrophage-generated O2-. This amplification was specifically reduced to the steady-state level by antioxidants, and further to the basal level by anti-CD4 antibodies, indicating the specificity of O2- for enhancing HIV-1 spread by cell-to-cell transmission.ELSEVIER SCIENCE BV, 1993年09月, FEBS LETTERS, 331(1-2) (1-2), 182 - 186, 英語[査読有り]研究論文（学術雑誌）
- Amplification of superoxide (O2-) generation by HIV-1 infection was examined in two human myeloid-monocytic cell lines. The level of O2- generation in HL-60 after infection became significantly higher than that of the steady-state. A similar phenomenon was also shown in U937, but only after acquisition of O2- generation ability by differentiation to macrophages. By means of the NADPH oxidase-coupled response in infected cells, we reconstituted the O2-generating machinery in cell-free system. The results suggested that cytosolic factor(s) exerted by infection might be responsible for the amplification of O2- generation. Thus, HIV-1 infection could elevate the level of oxidative stress in macrophages which might play an important role in disease progression.ELSEVIER SCIENCE BV, 1993年07月, FEBS LETTERS, 326(1-3) (1-3), 232 - 236, 英語[査読有り]研究論文（学術雑誌）
- 2021年, 日本ウイルス学会学術集会プログラム・予稿集(Web), 68thアメントフラボンのB型肝炎ウイルス感染阻害活性について
- (一社)日本臨床微生物学会, 2020年12月, 日本臨床微生物学会雑誌, 31(Suppl.1) (Suppl.1), 201 - 201, 日本語インドネシアの食肉から分離された薬剤耐性Salmonella entericaの遺伝子解析
- ELSEVIER SCIENCE BV, 2017年09月, INTERNATIONAL JOURNAL OF ANTIMICROBIAL AGENTS, 50, S82 - S82, 英語Mitochondrial DNA depletion among HIV-infected patients at Sanjiwani Hospital, Gianyar, Bali研究発表ペーパー・要旨（国際会議）
- 2015年04月, 最新医学, 74(4) (4), 745 - 743, 日本語神戸大学インドネシア拠点のあゆみと実績[査読有り][招待有り]その他
- The Center for Infectious Diseases (CID), Kobe University Graduate School of Medicine, has led an Asia-related medical research program for over 50 years. The Japan Initiative for Global Research Network on Infectious Diseases (J-GRID) established the Indonesia-Kobe University Collaborative Research Center for Emerging and Reemerging Infectious Diseases (CRC-FRID), which is staffed by Japanese researchers from the CID, Kobe University Graduate School of Medicine, and Indonesian researchers from the Institute of Tropical Disease (ITD) of Airlangga University, Surabaya, Indonesia. There they focus on five disease types – influenza, infectious hepatitis, dengue fever, HIV/AIDS, and infectious diarrheal diseases – in collaborative research. This paper summarizes research results for these 5 diseases as published in previous papers.Fuji Technology Press, 2014年10月01日, Journal of Disaster Research, 9(5) (5), 828 - 835, 英語書評論文,書評,文献紹介等
- ELSEVIER SCI LTD, 2012年06月, INTERNATIONAL JOURNAL OF INFECTIOUS DISEASES, 16, E191 - E191, 英語研究発表ペーパー・要旨（国際会議）
- ELSEVIER SCI LTD, 2012年06月, INTERNATIONAL JOURNAL OF INFECTIOUS DISEASES, 16, E193 - E194, 英語研究発表ペーパー・要旨（国際会議）
- ELSEVIER SCI LTD, 2012年06月, INTERNATIONAL JOURNAL OF INFECTIOUS DISEASES, 16, E194 - E194, 英語研究発表ペーパー・要旨（国際会議）
- 日本エイズ学会, 2011年11月, 日本エイズ学会誌, 13(4) (4), 482 - 482, 日本語シクロフィリンAをターゲットとする新たな候補を見つけるための分子ドッキング法の検討(Investigation of molecular docking method in order to find novel candidates targeting cyclophilin A)
- 2011年05月, ANTIVIRAL RESEARCH, 90(2) (2), A76 - A77[査読有り]
- (公社)日本薬学会, 2011年03月, 日本薬学会年会要旨集, 131年会(4) (4), 126 - 126, 日本語CypAをターゲットとする抗HIV-1低分子化合物のin Silico探索
- Background: Protease (PR) inhibitors (PIs) were designed against subtype B virus of human immunodeficiency virus type 1 (HIV-1), but believed to retain its activity against most of the other subtypes. CRF01_AE PR (AE-PR) contains background mutations that are presumed to alter the drug susceptibility of PR. In addition, amino acid variations found in HIV-1 Gag potentially affect the drug susceptibility or catalytic efficiency of PR. Methods: We studied the impact of naturally occurring amino acid substitutions found in AE-PR and CRF01_AE Gag (AE-Gag) on the drug susceptibility of PR to 9 currently available Pis, using the pNL4-3-derived luciferase reporter virus containing AE-Gag and/or AE-PR genes derived from drug treatment-naive, HIV-1-infected Thai patients. Results: Sequencing analysis revealed that several mutations were detected in deduced amino acid sequences of AE-PR and AE-Gag genes, as compared to these genes of pNL4-3. Drug susceptibility tests revealed that AE-PR showed a variety of susceptibilities to 9 Pis compared with pNL4-3 PR. In addition, AE-Gag significantly reduced the drug susceptibility of AE-PR and pNL4-3 PR. Conclusion: Our results suggest that amino acid variations in AE-PR and AE-Gag play roles in determining the drug susceptibility of CRF01_AE viruses to PIs.LIPPINCOTT WILLIAMS & WILKINS, 2009年11月, JAIDS-JOURNAL OF ACQUIRED IMMUNE DEFICIENCY SYNDROMES, 52(3) (3), 320 - 328, 英語
- Axin1, a regulator of Wnt signaling, was previously identified as playing a negative role in the late phase of human immunodeficiency virus type 1 (HIV-1) replication in HeLa-derived J111 cells. In this report, we studied the molecular mechanism of how Axin1 regulates HIV-1 replication. HIV-1 transactivator, Tat-dependent viral reporter gene expression was enhanced in J111 cells transfected with small interfering RNA (siRNA) against Axin1. In addition, viral transcription was upregulated in J111 cells transfected with siRNA against Axin1. In contrast, HIV-1 gene expression was not enhanced by transfecting HeLa cells with siRNA against Axin1. The expression levels of T cell factor-4 (TCF4) and beta-catenin were higher in J111 than HeLa cells. In addition, siRNAs against TCF4 and beta-catenin inhibited the Axin1 siRNA-dependent enhancement of HIV-1 gene expression in J111 cells. These results suggest that Axin1 plays a negative role in HIV-1 transcription through the Wnt signaling pathway in J111 cells under normal cell culture conditions.MARY ANN LIEBERT INC, 2009年10月, AIDS RESEARCH AND HUMAN RETROVIRUSES, 25(10) (10), 1005 - 1011, 英語
- CRF01_AE is a major subtype of human immunodeficiency virus type 1 (HIV-1) circulating in Southeast Asia, including Thailand. We performed genotypic studies on HIV-1 CRF01_AE protease (PR) and reverse transcriptase (RT) derived from plasma samples from drug treatment-naive patients residing in central Thailand. Direct sequencing of amplified CRF01_AE PR and RT genes revealed that drug resistance-associated as well as background mutations were frequently detected in CRF01_AE PR. In contrast, although several background mutations were detected, no drug resistance-associated mutations were observed in CRF01_AE RT. Antiretroviral drugs, including PR inhibitors, have been increasingly used in Thailand. To achieve effective antiretroviral therapy, we propose that it is important to reveal the prevalence of drug resistance-associated mutations among drug treatment-naive patients by further surveillance studies.MARY ANN LIEBERT INC, 2009年06月, AIDS RESEARCH AND HUMAN RETROVIRUSES, 25(6) (6), 625 - 631, 英語
- Human immunodeficiency virus type I (HIV-1) env genes were cloned from blood samples of HIV-1-infected Thai patients, and 35 infectious CRF01_AE envelope glycoprotein (Env)-recombinant viruses were established. In this report, we examined the neutralization susceptibility of these viruses to human monoclonal antibodies, 2G12, IgG1 b12, 2175 and 4E10, pooled patient plasma, coreceptor antagonists and fusion inhibitor, T-20. The neutralization susceptibility of CRF01_AE Env-recombinant viruses to 2F5, 4E10, patient plasma, coreceptor antagonists and T-20 varied, while most viruses showed low susceptibility to 2G12 and IgG1 b12. Several dual-tropic viruses showed lower susceptibility to 2F5 and 4E10 than CXCR4- or CCR5-tropic viruses. Neutralization susceptibility of the CRF01_AE Env-recombinant virus to pooled patient plasma was negatively correlated with the length of the V1/V2 region or the number of potential N-linked glycosylation sites in conserved regions of gp120. No correlation was found between the coreceptor usage and neutralization susceptibility of the virus to T-20, whereas several dual-tropic viruses showed higher susceptibility to coreceptor antagonists than CXCR4- or CCR5-tropic viruses. We propose that these CRF01_AE Env-recombinant viruses are useful to further study the molecular mechanism of the susceptibility of CRF01_AE Env to neutralizing antibodies and viral entry inhibitors. (C) 2008 Elsevier Masson SAS. All rights reserved.ELSEVIER SCIENCE BV, 2009年03月, MICROBES AND INFECTION, 11(3) (3), 334 - 343, 英語
- CRF01_AE is a major subtype of human immunodeficiency virus type 1 (HIV-1) circulating in Southeast Asia, including Thailand. HIV-1 env genes were amplified by polymerase chain reaction from blood samples of HIV-1-infected patients residing in Thailand in 2006, and cloned into the pNL4-3-derived reporter viral construct. Generated envelope protein (Env)-recombinant virus was examined for its infectivity, and then 35 infectious CRF01_AE Env-recombinant viruses were selected. Sequencing analysis revealed that the interclone variation of the deduced amino acid sequences was higher in CRF01_AE env genes isolated in 2006 than in those isolated in the early 1990s, suggesting that env gene variation has been increasing gradually among CRF01_AE viruses prevalent in Thailand. We also examined the characteristics of the deduced amino acid sequences of 35 CRF01_AE env genes. Our results may provide useful information to help in better understanding the genotype of env genes of CRF01_AE viruses currently circulating in Thailand.MARY ANN LIEBERT INC, 2009年02月, AIDS RESEARCH AND HUMAN RETROVIRUSES, 25(2) (2), 229 - 236, 英語
- ELSEVIER SCIENCE BV, 2008年05月, ANTIVIRAL RESEARCH, 78(2) (2), A42 - A42, 英語[査読有り]研究発表ペーパー・要旨（国際会議）
- (公社)日本薬学会, 2008年03月, 日本薬学会年会要旨集, 128年会(2) (2), 53 - 53, 日本語HIV膜融合阻害剤C34に関する構造活性相関研究
- The transfection of human cells with siRNA against adapter-related protein complex 2 alpha 1 subunit (AP2 alpha) was revealed to significantly upregulate the replication of human immunodeficiency virus type 1 (HIV-1). This effect was confirmed by cell infection with vesicular stomatitis virus G protein-pseudotyped HIV-1 as well as CXCR4-tropic and CCR5-tropic HIV-1. Viral adsorption, viral entry and reverse transcription processes were not affected by cell transfection with siRNA against AP2 alpha. In contrast, viral nuclear translocation as well as the integration process was significantly up-regulated in cells transfected with siRNA against AP2 alpha. Confocal fluorescence microscopy revealed that a subpopulation of AP2 alpha was not only localized in the cytoplasm but was also partly co-localized with lamin B, importin beta and Nup 153, implying that AP2 alpha negatively regulates HIV-1 replication in the process of nuclear translocation of viral DNA in the cytoplasm or the perinuclear region. We propose that AP2 alpha may be a novel target for disrupting HIV-1 replication in the early stage of the viral life cycle. (C) 2007 Elsevier Inc. All rights reserved.ACADEMIC PRESS INC ELSEVIER SCIENCE, 2008年03月, VIROLOGY, 373(1) (1), 171 - 180, 英語
- 日本エイズ学会, 2007年11月, 日本エイズ学会誌, 9(4) (4), 405 - 405, 日本語HIV gp41の蛋白質間相互作用計算によるアミノ酸残基と膜融合阻害活性との相関
- We performed the screening to find the novel host factors affecting human immunodeficiency virus type-1 (HIV-1) replication using the siRNA mini-library consisted with 257 siRNAs directed against cellular genes. J111 cells, a human acute monocytic leukemia cell line, were transfected with individual siRNA, followed by either infected or transfected with the HIV-1 molecular clone with luciferase reporter gene in 96-well plate format. The results showed that six siRNAs significantly enhanced the HIV-1 replication in J111 cells, indicating that the target cellular genes of those siRNAs may negatively regulate HIV-1 replication in normal cell culture condition. We also discuss the possible mechanisms by which those cellular proteins regulate viral replication. (c) 2007 Elsevier Inc. All rights reserved.ACADEMIC PRESS INC ELSEVIER SCIENCE, 2007年08月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 359(3) (3), 729 - 734, 英語
- 2007年, 情報計算化学生物学会大会予稿集, 2007HIV-1 gp41 N末端7回反復配列と膜融合阻害剤C34との間の6ヘリックス束モデルの計算アラニン走査
- 2007年01月, 英語Computational Studies on Relationship with Protein-Protein Interaction Energy of HIV-1 gp41 and Its Inhibitory Activity of Membrane Fusion
- (公社)日本生化学会, 2002年08月, 生化学, 74(8) (8), 1088 - 1088, 日本語成熟脳中のApaf-1阻害因子の精製
- ヒト免疫不全ウイルス(HIV)やその他のレトロウイルスの増殖機構におけるポリADPリボース合成酵素の役割AIDSの原因ウイルスであるヒト免疫不全ウイルス(HIV)やその他のレトロウイルスの生活環の複数の増殖ステップに，ポリADPリボース合成酵素(PARP)が制御因子として関わっている。その制御機構を明らかにすることは，新たなAIDS治療薬を開発する手がかりとなりうるものである。奈良医学会, 2002年04月30日, Journal of Nara Medical Association, 53(2) (2), 91 - 99, 日本語
- (公社)日本生化学会, 2001年08月, 生化学, 73(8) (8), 983 - 983, 日本語PARP欠損細胞における熱ショック誘導アポトーシスの亢進
- (公社)日本生化学会, 2001年08月, 生化学, 73(8) (8), 1033 - 1033, 日本語Tat依存的なHIV-1転写機構におけるポリADPリボース合成酵素の役割について
- (公社)日本生化学会, 2000年08月, 生化学, 72(8) (8), 935 - 935, 日本語Apaf-1依存性のcaspase活性化 胸腺,脳,肝臓組織における比較
- (公社)日本生化学会, 2000年08月, 生化学, 72(8) (8), 935 - 935, 日本語アポトーシスならびにネクローシスにおけるポリADPリボース合成酵素の相反する役割
- (公社)日本生化学会, 2000年08月, 生化学, 72(8) (8), 935 - 935, 日本語成熟脳におけるApaf-1経由のアポトーシス経路の停止
- (公社)日本生化学会, 1999年08月, 生化学, 71(8) (8), 820 - 820, 日本語ポリ(ADP-リボース)合成酵素によるNF-κBの調節機構
- (公社)日本生化学会, 1999年08月, 生化学, 71(8) (8), 822 - 822, 日本語牛の脳及び胸腺のcaspaseと活性化因子
- 奈良医学会, 1998年12月, Journal of Nara Medical Association, 49(6) (6), 495 - 495, 日本語牛胸腺抽出液におけるcaspaseの活性化機構
- (公社)日本生化学会, 1998年08月, 生化学, 70(8) (8), 787 - 787, 日本語アポトーシスの分子機構 牛胸腺抽出液中のcaspase-3活性化因子
- (公社)日本生化学会, 1998年08月, 生化学, 70(8) (8), 1050 - 1050, 日本語U937由来サブクローンのアポトーシス誘導感受性の解析
- 北海道大学, 1998年, Collected papers from the Institute of Immunological Science Hokkaido University, 21, 80 - 88, 英語Dependence of host cell cycle for activation of human immunodeficiency virus type 1 gene expression from latency
- 北海道大学, 1998年, Collected papers from the Institute of Immunological Science Hokkaido University, 21, 101 - 108, 英語A specific T-cell subset with CD4+/CD38- markers derived from HIV-1 cariers induses apoptosis in healthy donor-derived T-lymphocytes.
- FEDERATION AMER SOC EXP BIOL, 1997年07月, FASEB JOURNAL, 11(9) (9), A981 - A981, 英語Persistent infection of U937 cells with HIV-1 blocks CPP32-dependent apoptotic pathway in the cells研究発表ペーパー・要旨（国際会議）
- 北海道大学, 1997年, Collected papers from the Institute of Immunological Science Hokkaido University, 20, 46 - 59, 英語Insertions, duplications and substitutions in restricted gp90 regions of equine infectious anemia virus occurred during febrile episodes in an experimentally infected horse.
- 北海道大学, 1997年, Collected papers from the Institute of Immunological Science Hokkaido University, 20, 17 - 23, 英語Protease-defective, gp120-containing human immunodeficiency virus type 1 particles induce apoptosis more efficiently than does wild-type virus or recombinant gp120 protein in healthy donor-derived peripheral blood T cells.
- 北海道大学, 1997年, Collected papers from the Institute of Immunological Science Hokkaido University, 20, 60 - 67, 英語A chain section containing epitopes for cytotoxic T, B and helper T cells within a highly conserved region found in the human immunodeficiency virus type 1 Gag protein.
- 北海道大学, 1997年, Collected papers from the Institute of Immunological Science Hokkaido University, 20, 125 - 130, 英語Superoxide generation by monocytes following infection with human cytomegalovirus.
- 北海道大学, 1996年, Collected papers from the Institute of Immunological Science Hokkaido University, 19, 74 - 77, 英語E Intracellular glutathione as a possible direct blocker of HIV type 1 reverse transcription.
- 北海道大学, 1995年, Collected papers from the Institute of Immunological Science Hokkaido University, 18, 31 - 42, 英語Stimulation of human immunodeficiency virus type 1 infected cells with superoxide enhances the chemotactic motile response of CD4+ human cells: implication for virus transmission by cell-to-cell interaction.
- 北海道大学, 1995年, Collected papers from the Institute of Immunological Science Hokkaido University, 18, 43 - 55, 英語High susceptibility of U937-derived subclone to infection with human immunodeficiency virus type 1 is correlated with virus-induced celldifferentiation and superoxide generation.
- 北海道大学, 1995年, Collected papers from the Institute of Immunological Science Hokkaido University, 18, 69 - 76, 英語Poly(ADP-ribose) polymerase activity in various U937 cell subclones with different susceptibility to HIV-1 infection : Its dramatic decre ase following persistent virus infection.
- MARY ANN LIEBERT INC PUBL, 1995年, AIDS RESEARCH AND HUMAN RETROVIRUSES, 11, S117 - S117, 英語EXTRACELLULAR NEF PROTEIN REGULATES ACTIVATION OF HIV-INFECTION FROM LATENTLY INFECTED-CELLS研究発表ペーパー・要旨（国際会議）
- MARY ANN LIEBERT INC PUBL, 1995年, AIDS RESEARCH AND HUMAN RETROVIRUSES, 11, S112 - S112, 英語APOPTOTIC CELL-LYSIS INDUCED BY DEFECTIVE, HIV PROTEASE-DEFICIENT PARTICLES研究発表ペーパー・要旨（国際会議）
- 北海道大学, 1994年, Collected papers from the Institute of Immunological Science Hokkaido University, 17, 85 - 94, 英語In vivo induction of human immunodeficiency virus type 1-specific cytotoxic T lymphocytes and delayed-type hypersensitivity by a @@S ' 23-amino acid peptide from the highly conserved region in major @@S ' core protein p24
- 北海道大学, 1994年, Collected papers from the Institute of Immunological Science Hokkaido University, 17, 71 - 76, 英語Cytotoxic T lymphocyte response in mice induced by a recombinant BCG Vaccination which produces an extracellular α antigen that @@S ' fused with the human immunodeficiency virus type 1 envelope immuno @@S dominant domain in the V3 loop.
- 北海道大学, 1994年, Collected papers from the Institute of Immunological Science Hokkaido University, 17, 59 - 70, 英語The carboxyl-terminal region of HIV-1 Nef protein is a cell surface domain that can interact with CD4+ T cells
- 北海道大学, 1993年, Collected papers from the Institute of Immunological Science Hokkaido University, 16, 80 - 86, 英語Superoxide enhances the spread of HIV-1 infection by cell-to-cell
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- 第67回日本ウイルス学会学術集会, 2019年10月PMLタイプJCポリオーマウイルスの培養細胞における増殖
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- 第33回近畿エイズ研究会, 2019年, 日本語ネパール大地震がHIV感染者の服薬アドヒアランス、メンタルヘルスと薬剤耐性変異の出現におよぼす影響について
- 第４１回日本分子生物学会年会, 2018年11月, 英語, 日本分子生物学会, 横浜, 国内会議CRISPR/Cas9 System Targeting Regulatory Genes of HIV-1 Inhibits Viral Replication in Infected T-Cell Culturesポスター発表
- 第22回日本ワクチン学会学術集会, 2018年, 日本語, 国内会議デングウイルスの感染増強を起こさない日本脳炎ワクチンの開発口頭発表（一般）
- 第53回日本脳炎ウイルス生態学研究会, 2018年, 日本語, 国内会議デングウイルスに対する感染増強抗体を誘導しない日本脳炎ワクチンの開発口頭発表（一般）
- 第66回日本ウイルス学会学術集会, 2018年, 日本語, 国内会議prM領域を改変したデングウイルス2型DNAワクチンの評価口頭発表（一般）
- 第24回 トガフラビペスチウイルス研究会, 2017年, 日本語, 国内会議デングウイルスの増殖に関連する宿主細胞因子の検索口頭発表（一般）
- 第24回 トガフラビペスチウイルス研究会, 2017年, 日本語, 国内会議デングウイルスに対する血清型特異的抗体の樹立口頭発表（一般）
- 第65回日本ウイルス学会, 2017年, 日本語, 国内会議Nearly full-length sequencing of HIV-1 subtype B in Indonesiaポスター発表
- 第31回近畿エイズ研究会学術集会, 2017年, 日本語, 国内会議IV-1 CRF01_AE株がgp120 CD4結合部位を認識する単クローン抗体VRC01に対して中和抵抗性を示す分子機構口頭発表（一般）
- Asia-Africa Research Forum on Emerging and Reemerging Infections 2014, 2017年, 英語, 国際会議High prevalence of HIV-1 CRF01_AE viruses among female commercial sex workers residing in Surabaya, Indonesia口頭発表（一般）
- 第65回日本ウイルス学会, 2017年, 英語, 国内会議Genotypic characterization of human immunodeficiency virus type 1 derived from infected individuals residing in Bali, Indonesiaポスター発表
- 第65回日本ウイルス学会, 2017年, 日本語, 国内会議Characterization of HIV-1 env genes derived from recently infected Indonesian individualsポスター発表
- 第65回日本ウイルス学会, 2017年, 英語, 国内会議Appearance of drug resistance-associated mutations in human immunodeficiency virus type 1 protease and reverse transcriptase derived from infected individuals in Papua, Indonesiaポスター発表
- 第65回日本ウイルス学会, 2017年, 日本語, 国内会議Analysis of an epitope located near the glycosylation site on the domain II of dengue virus type 1 E proteinポスター発表
- 第51回 日本脳炎ウイルス生態学研究会, 2016年, 日本語, 国内会議デングウイルス1型望月株に対して強力な中和活性を示すマウスモノクローナル抗体のエピトープ解析口頭発表（一般）
- 第23回 トガフラビペスチウイルス研究会, 2016年, 日本語, 国内会議インドネシアのデング患者血より作製された 抗デングウイルスヒト型モノクローナル抗体の性状解析口頭発表（一般）
- Asia-Africa Research Forum on Emerging and Reemerging Infections 2014, 2016年, 英語, 国際会議Molecular epidemiology of Dengue Virus in 4 cities of East Java, Indonesia口頭発表（一般）
- 第64回 日本ウイルス学会, 2016年, 英語, 国内会議Characterization of human monoclonal antibodies against dengue virus generated from Indonesian dengue patients口頭発表（一般）
- Asia-Africa Research Forum on Emerging and Reemerging Infections 2014, 2015年, 英語, 国際会議Generation of human monoclonal antibody using peripheral blood lymphocytes from dengue patient in Indonesiaポスター発表
- 第63回 日本ウイルス学会学術集会, 2015年, 日本語, 国内会議Divergence of dengue virus 2 Cosmopolitan genotype associated with two predominant serotype shifts between 1 and 2 in Surabaya, Indonesiaポスター発表
- 第28回日本エイズ学会学術集会・総会, 2014年12月, 日本語, 日本エイズ学会, 大阪, 国内会議東南アジアで流行するHIVの特徴[招待有り]シンポジウム・ワークショップパネル（指名）
- 第28回日本エイズ学会学術集会・総会, 2014年12月, 日本語, 日本エイズ学会, 大阪, 国内会議ビオチン化PIの分子設計と活性プロテアーゼの同定口頭発表（一般）
- 第28回日本エイズ学会学術集会・総会, 2014年12月, 日本語, 日本エイズ学会, 大阪, 国内会議インドネシア・スラバヤ市の性産業従事者におけるHIV流行口頭発表（一般）
- Joint International Tropical Medicine Meeting (JITMM 2014) and Food borne Parasites and Zoonoses 8 (FBPZ8), 2014年12月, 英語, Mahidol University, バンコク, タイ, 国際会議Dengue virus infection-neutralizing and enhancing antibody responses in central Thai populations against Indonesian and Thai strains.口頭発表（一般）
- 第62回日本ウイルス学会学術集会, 2014年11月, 日本語, 日本ウイルス学会, 横浜, 国内会議ヒト神経芽細胞腫でのTNF-αによるJCウイルスDNA複製の促進口頭発表（一般）
- 第62回日本ウイルス学会学術集会, 2014年11月, 日本語, 日本ウイルス学会, 横浜, 国内会議インドネシアの薬剤未治療HIV-1患者における薬剤耐性変異の解析ポスター発表
- 第21回トガ・フラビ・ペスチウイルス研究会, 2014年11月, 日本語, 国立感染症研究所, 横浜, 国内会議インドネシアのデング患者血を用いた抗デングウイルスヒト型モノクローナル抗体の樹立口頭発表（一般）
- 第28回近畿エイズ研究会学術集会, 2014年06月, 日本語, 近畿エイズ研究会, 大阪, 国際会議インドネシア・スラバヤ市に流行するHIV-1の遺伝子解析口頭発表（一般）
- 日本薬学会年会, 2014年03月, 日本語, 国内会議ビオチン化アスパラギン酸プロテアーゼ阻害剤を用いたHIVプロテアーゼ研究口頭発表（一般）
- Asia-Africa Research Forum on Emerging and Reemerging Infections 2014, 2014年, 英語, 国際会議Appearance of drug resistance-associated mutations in human immunodeficiency virus type 1 protease, reverse transcriptase and integrase derived from drug-naïve Indonesian patientsポスター発表
- Asian-African Research Forum on Emerging and Reemerging infections 2014, 2014年01月, 英語, 国際会議Molecular epidemiology of dengue virus in 4 cities of east Java, Indonesia.口頭発表（一般）
- Asian-african research forum on emerging and reemerging infections 2014, 2014年01月, 英語, MEXT, J-GRID, 仙台, 国際会議Molecular epidemiology of Dengue Virus in 4 cities of East Java, Indonesia口頭発表（一般）
- Asian-African Research Forum on Emerging and Reemerging infections 2014, 2014年01月, 英語, 国際会議High prevalence of HIV-1 CRF01_AE viruses among female commercial sex workers residing in Surabaya, Indonesia.口頭発表（一般）
- Asian-african research forum on emerging and reemerging infections 2014, 2014年01月, 英語, MEXT, J-GRID, 仙台, 国際会議High prevalence of HIV-1 CRF01_AE viruses among female commercial sex workers residing in Surabaya, Indonesia.口頭発表（一般）
- Asian-african research forum on emerging and reemerging infections 2014, 2014年01月, 英語, MEXT, J-GRID, 仙台, 国際会議Generation of human monoclonal antibody using peripheral blood lymphocytes from dengue patient in Indonesiaポスター発表
- Asian-African Research Forum on Emerging and Reemerging infections 2014, 2014年01月, 英語, 国際会議Generation of human monoclonal antibody using peripheral blood lymphocites from dengue patient in Indonesia.ポスター発表
- Asian-African Research Forum on Emerging and Reemerging infections 2014, 2014年01月, 英語, 国際会議Appearance of drug resistance-associated mutations in human immunodeficiency virus type 1 protease, reverse transcriptase and integrase derived from drug-naïve Indonesian patients.ポスター発表
- Asian-african research forum on emerging and reemerging infections 2014, 2014年01月, 英語, MEXT, J-GRID, 仙台, 国際会議Appearance of drug resistance-associated mutations in human immunodeficiency virus type 1 protease, reverse transcriptase and integrase derived from drug-naïve Indonesian patientsポスター発表
- 日本環境動物昆虫学会年次大会, 2013年11月, 日本語, 国内会議蚊が関係する疾病[招待有り]シンポジウム・ワークショップパネル（指名）
- トガ・フラビ・ペスチウイルス研究会, 2013年11月, 日本語, 国内会議スラバヤ市における2008年から2013年までのデングウイルス分子疫学口頭発表（一般）
- 第20回 トガ・フラビ・ペスチウイルス研究会, 2013年11月, 日本語, 国立感染症研究所, 神戸, 国内会議インドネシア国スラバヤ市における2008年から2013年までのデングウイルス分子疫学口頭発表（一般）
- 第61回日本ウイルス学会学術集会, 2013年11月, 日本語, 日本ウイルス学会, 神戸, 国内会議インドネシア・スラバヤの性産業従事者を対象としたHIV及び肝炎ウイルスの疫学調査ポスター発表
- 日本ウイルス学会学術集会, 2013年11月, 日本語, 国内会議インドネシア・スラバヤの性産業従事者を対象としたHIVおよび肝炎ウイルスの疫学調査ポスター発表
- 第61回日本ウイルス学会学術集会, 2013年11月, 英語, 日本ウイルス学会, 神戸, 国内会議Tetravalent dengue DNA vaccine containing premembrane and envelope genes of dengue virus isolates in Indonesia.口頭発表（一般）
- 日本ウイルス学会学術集会, 2013年11月, 英語, 国内会議Tetravalent dengue DNA vaccine containing premembrane and envelope genes of dengue virus isolates in Indonesia口頭発表（一般）
- 日本ウイルス学会学術集会, 2013年11月, 日本語, 国内会議PARP-1阻害剤のin vitroにおけるJCウイルス増殖抑制効果について口頭発表（一般）
- 日本ウイルス学会学術集会, 2013年11月, 日本語, 国内会議HIV-1 CRF01_AE株がgp120 CD4結合部位を認識する単クローン抗体に対して中和抵抗性を示す分子機構口頭発表（一般）
- 第61回日本ウイルス学会学術集会, 2013年11月, 日本語, 日本ウイルス学会, 神戸, 国内会議Antiviral activity of crude extracts from Japanese medical plants against dengue virus type-2.ポスター発表
- 日本ウイルス学会学術集会, 2013年11月, 英語, 国内会議Antiviral activity of crude extracts from Japanese medical plants against dengue virus type-2ポスター発表
- 近畿エイズ研究会学術集会, 2013年06月, 日本語, 国内会議インドネシア・スラバヤ市の性産業従事者を対象としたHIVと肝炎ウイルスの疫学調査口頭発表（一般）
- 第27回近畿エイズ研究会学術集会, 2013年06月, 日本語, 近畿エイズ研究会, 大阪, 国内会議インドネシア・スラバヤ市の性産業従事者を対象としたHIVと肝炎ウイルスの疫学調査口頭発表（一般）
- 近畿エイズ研究会学術集会, 2013年06月, 日本語, 国内会議TNF-αによるJCウイルスのLarge T抗原の発現促進口頭発表（一般）
- Asia-Africa Research Forum on Emerging and Reemerging Infections 2013, 2013年, 英語, 国際会議Phylogenetic and Molecular Clock Analysis of Dengue Virus Strains Isolated in Surabaya and Sidoarjo, Indonesia, during 2011-2012ポスター発表
- Asia-Africa Research Forum on Emerging and Reemerging Infections 2013, 2013年, 英語, 国際会議Awareness of using Ringer’s Lactate Solution in Dengue Virus Infection can Induce Severityポスター発表
- Asian-African Research Forum on Emerging and Reemerging infections 2013, 2013年01月, 英語, 国際会議Phylogenetic and Molecular Clock Analysis of Dengue Virus Strains Isolated in Surabaya and Sidoarjo, Indonesia, during 2011-2012.ポスター発表
- Asian-African Research Forum on Emerging and Reemerging Infections (AARF) 2013, 2013年01月, 英語, Ministry of Education, Culture, Sports, Science and Technology. Japan Initiative for Global Research Network on Infectious Diseases., 東京, Dengue virus (DENV) is transmitted to humans by the bite of an infected Aedes mosquito. Once infected, the mosquito remains infected for life, transmitting the virus to susceptible individuals while probing and feeding. The transmission cycle between humans and mosquitoes is a main mechanism for the maintenance of DENV in a large human populations and abundant Aedes mosquito po, 国際会議Phylogenetic and Molecular Clock Analysis of Dengue Virus Strains Isolated in Surabaya and Sidoarjo, Indonesia, during 2011-2012ポスター発表
- Asian-African Research Forum on Emerging and Reemerging infections 2013, 2013年01月, 英語, 国際会議Awareness of using Ringer’s lactate solution in dengue virus infection can induce severity.口頭発表（一般）
- Asian-African Research Forum on Emerging and Reemerging Infections (AARF) 2013, 2013年01月, 英語, Ministry of Education, Culture, Sports, Science and Technology. Japan Initiative for Global Research Network on Infectious Diseases., 東京, Dengue virus (DENV) infection is one of the important health problems in Indonesia, although the mortality rate has been decreased. Many cases with dengue shock syndrome and unusual manifestation of dengue infection are difficult to predict the earlier time for getting a good management. We made updated management of unusual manifestation in dengue infection for getting a bette, 国際会議Awareness of using Ringer's lactate solution in dengue virus infection can induce severity口頭発表（一般）
- 日本ウイルス学会学術集会, 2012年11月, 日本語, 国内会議タイにおけるJ-GRIDおよびSATREPS研究活動について[招待有り]シンポジウム・ワークショップパネル（指名）
- 第60回日本ウイルス学会学術集会, 2012年11月, 日本語, 日本ウイルス学会, 大阪, タイでおこなわれたJ-GRIDおよびSATREPS研究課題の概略を報告した, 国内会議タイにおけるJ-GRIDおよびSATREPS研究活動についてシンポジウム・ワークショップパネル（指名）
- 日本ウイルス学会学術集会, 2012年11月, 日本語, 国内会議シクロフィリンAとHIV-1 Gagの結合を阻害する新規低分子化合物の検索ポスター発表
- 第60回日本ウイルス学会学術集会, 2012年11月, 日本語, 日本ウイルス学会, 大阪, シクロフィリンAとHIV-1 Gagの結合を阻害する低分子化合物を解析して、有効な化合物を見いだした, 国内会議シクロフィリンAとHIV-1 Gagの結合を阻害する新規低分子化合物の検索ポスター発表
- 日本ウイルス学会学術集会, 2012年11月, 日本語, 国内会議HIV-1 Tatによる神経芽細胞腫でのJCウイルス増殖促進口頭発表（一般）
- 第60回日本ウイルス学会学術集会, 2012年11月, 日本語, 日本ウイルス学会, 大阪, HIV-1 Tatにより神経芽細胞腫におけるJCウイルス増殖促進機構を解析した, 国内会議HIV-1 Tatによる神経芽細胞腫でのJCウイルス増殖促進口頭発表（一般）
- 第26回近畿エイズ研究会学術集会, 2012年07月, 日本語, 近畿エイズ研究会, 神戸, HIV-1 TatによりJCウイルスの増殖が更新する分子機構を解析した, 国内会議HIV-1 TatのPML型JCウイルス増殖促進口頭発表（一般）
- 近畿エイズ研究会学術集会, 2012年07月, 日本語, 国内会議HIV-1 TatによるJCウイルス増殖促進機構の解明口頭発表（一般）
- 近畿エイズ研究会学術集会, 2012年07月, 日本語, 国内会議HIV-1 CRF01_AE株がCD4結合部位を認識する単クローン抗体に対して中和抵抗性を示す分子機構口頭発表（一般）
- 第26回近畿エイズ研究会学術集会, 2012年07月, 日本語, 近畿エイズ研究会, 神戸, HIV-1 CRF01_AE株がCD4結合部位を認識する単クローン抗体に対して中和抵抗性を示す分子機構として、外被タンパク質gp120のV2領域の1アミノ酸の役割を明らかにした, 国内会議HIV-1 CRF01_AE株がCD4結合部位を認識する単クローン抗体に対して中和抵抗性を示す分子機構口頭発表（一般）
- 15th International Congress on Infectious Diseases, 2012年06月, 英語, 国際会議Susceptibility of HIV-1 CRF01_AE viruses to a CD4-binding site monoclonal antibody, IgG1 b12.ポスター発表
- 15th International Congress on Infectious Diseases, 2012年06月, 英語, 国際会議In search for a new anti-HIV-1 drug through inhibition of CA-CypA interaction.ポスター発表
- 15th International Congress on Infectious Diseases, 2012年06月, 英語, 国際会議Comparison of anti-HIV-1 neutralizing activity between the plasma derived from HIV-1 infected, slow and rapid progressors.ポスター発表
- Indonesia-Japan-Thailand Joint Forum on infectious Diseases 2012, 2012年, 英語, 国際会議Some dengue patient sera or monoclonal antibodies exhibit varying focus sizes in a dengue virus infection-enhancing antibody assay system口頭発表（一般）
- Indonesia-Japan-Thailand Joint Forum on infectious Diseases 2012, 2012年, 英語, 国際会議Sero-epidemiology of HIV-1infection and HBV, HCV co-infection among commercial sex worker in Surabaya, Indonesia口頭発表（一般）
- Indonesia-Japan-Thailand Joint Forum on infectious Diseases 2012, 2012年, 英語, 国際会議Genotypic study of HIV-1 derived from infected patients and commercial sex workers residing in Surabaya, Indonesia口頭発表（一般）
- Asian-African Research Forum on Emerging and Reemerging infections 2012, 2012年01月, 英語, 国際会議Evolution of HIV-1 CRF01_AE env gene in Thai patients.ポスター発表
- Asian-African Research Forum on Emerging and Reemerging infections 2012, 2012年01月, 英語, 国際会議Anti-HIV-1 humoral immune responses in HIV-1-infected Thai patients.口頭発表（一般）
- 日本薬学会年会, 2011年03月, 日本語, 国内会議CypAをターゲットとする抗HIV-1低分子化合物のin Silico探索口頭発表（一般）
■ 共同研究・競争的資金等の研究課題
研究シーズ
■ 研究シーズ- ヒト病原性ウイルスの研究シーズカテゴリ：ライフサイエンス, 自然科学一般研究キーワード：デング, エイズウイルス, SARS-CoV-2, COVID-19研究の背景と目的：新興感染症が次々と出現しています。特にヒト病原性ウイルスによる新たな感染症が数年おきに出現して公衆衛生上の問題となっています。ウイルス複製機構の詳細な解析やウイルス感染に対する宿主の免疫応答の解析は、新規の抗ウイルス薬やワクチン開発に欠かせない基礎研究と考えます。研究内容：これまで、エイズウイルス(HIV)、デングウイルスを含む蚊媒介感染症の原因となるフラビウイルス、最近ではCOVID-19の原因となるSARS-CoV-2などのヒト病原性ウイルスを対象として、ウイルス複製機構の解析、ウイルス複製に影響を及ぼす宿主細胞タンパク質の探索研究や、ウイルスタンパク質が感染細胞の性状に及ぼす影響などに関する基礎研究を行っています。また、抗ウイルス活性物質を探索するためのスクリーニング系の開発研究、ウイルスに対する中和抗体の活性測定系の開発研究、新規診断法の開発研究なども行っています。期待される効果や応用分野：ウイルス感染症の予防、診断、治療に役立つ情報の蓄積を目指しています。関係する業績：Kotaki T, Xie X, Shi P-Y, Kameoka M. A PCR amplicon–based SARS-CoV-2 replicon for antiviral evaluation. Scientific Reports 11(1):2229, 2021. 最近、SARS-CoV-2に対する抗ウイルス活性物質のスクリーニング系に応用可能なRNAレプリコンを開発しました。
mkameoka
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