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HIV/AIDS vaccine development： are we walking out from the dark？

Chinese Medical Journal 2010年第23期123卷 3489-3494页

作者： wan yan-min wanG You-chun XU Jian-qing Department of Scientific Research Shanghai Public HealthClinical Center Institutes of Biomedical Sciences Key Laboratoryof Medical Molecular Virology of Shanghai Medical College andInstitute of Medical Microbiology Fudan University Shanghai201508 China National Institute for the Control of-Pharmaceutical and BiologicalProducts Beijing 100050 China State Key Kaboratory for Infectious Disease Prevention andControl Chinese Center for Disease Control and PreventionBeijing 102206 China

The need for AIDS vaccine has been emphasized by the increase of HIV-1 prevalence in sexual transmission which bridges the spreading of HIV-1 from high-risk population to other populations. After more than two-decade intensive effort on the AIDS vaccine development, it remains elusive whether and how an effective vaccine will be achieved. Recent data released from a phase Ⅲ trial in Thailand showed a partial protection might be accomplishable by（R）the ＂prime-boost＂combination of two vaccines： ALVAC（R） HIV vaccine （the prime）, and AIDSVAX（R） B/E vaccine （the boost）.1 This unprecedented large clinical trial observed that the prime-boost combination lowered the rate of HIV infection by 31.2% compared to placebo based on the modified intent-to-treat population （n=51 vs. n=74,respectively; P=0.04）. However, debating on the efficacy interpretation of this trial arose among field ***, how to improve the efficacy will become the most important question to be tackled. Here we reviewed the recent publications and summarized the major progress achieved.

关键词： human immunodeficiency virus 1 vaccine innate immunity cross-reactive

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Chloramphenicol improved expression of recombinant cholera toxin B subunit in Escherichia coli and its adjuvanticity

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Chinese Medical Journal 2011年第17期124卷 2751-2755页

作者： XIE Xiao-yan wan yan-min ZHU Zhao-qin ZHANG Huan-xiang XU Jian-qing Shanghai Public Health Clinical Center Institutes of Biomedical Sciences Key Laboratory of Medical Molecular Virology of Ministry of Education Shanghai Medical College Fudan University Shanghai 201508 China Department of Cell Biology Medical College of Soochow University Jiangsu Province Key Laboratory of Stem Cell Research Suzhou Jiangsu 215123 China

Background Cholera toxin B subunit （CTB） was shown to be a potent adjuvant for protein immunogen, especially when inoculated through mucosal route. We aimed to optimize the expression approach for CTB and thereafter to determine the adjuvant effect on DNA vaccine. Methods Wild type CTB coding gene was amplified and cloned into prokaryotic expression vector pET-30a, and the recombinant CTB was expressed in the presence of different concentration of chloramphenicol and isopropyl β-D-thiogalactoside. Purified recombinant CTB was mixed with HIV-1 AE2f tat-rev-integrase-vif-nef fusion gene DNA vaccine and female BALB/c mice were vaccinated with a DNA priming-recombinant vaccinia vectored vaccine boosting regimen through intramuscular injection. Interferon γ （IFN-γ） enzyme-linked immunospot （Elispot） assay was used to read out the specific T-cell immunity. Results Chloramphenicol was essential for the efficient expression of recombinant CTB （rCTB） in pET-30a/BL21 （DE3） system and could be optimized at the concentration of 0.625 μg/ml in the presence of chloramphenicol. The purified rCTB could bind with GM1 efficiently. INF-γ Elispot data showed the T-cell response induced in CTB adjuvanted group （（734±240） spot forming cells/106 splenocytes） was higher than that induced by non-adjuvanted （（520±150） spot forming cells/10e splenocytes）, all responses against different antigens were enhanced in parallel. Conclusion CTB could be efficiently expressed in the presence of chloramphenicol and purified CTB is functional and capable of enhancincl the specific T cell responses elicited by DNA vaccine, the mechanism needs to be explored in the future.

关键词： HIV-1 DNA vaccine cholera toxin B subunit expression efficacy adjuvant

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Establishing a Th17 based mouse model for preclinical assessment of the toxicity of candidate microbicides

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Chinese Medical Journal 2010年第23期123卷 3381-3388页

作者： LI Liang-zhu yanG Yu YUAN Song-hua wan yan-min QIU Chao FENG yan-ling XU Jian-qing ZHANG Xiao-yan Research Center of Shanghai Public Health Clinical Center Institutes of Biomedical Sciences Fudan University Shanghai 201508 China Key Laboratory of Medical Molecular Virology & Institute of Medical Microbiology Shanghai Medical College of Fudan University Shanghai 200433 China State Key Laboratory for Infectious Disease Prevention andControl Chinese Center for Disease Control and PreventionBeijing 100050 China

Background To effectively block the invasion of human immunodeficiency virus （HIV）-1 on mucosal surface, vaginal anti-HIV-1 microbicides should avoid inflammatory responses and disruption of mucosa integrity because these will facilitate transepithelial viral penetration and replication. However, existing models fail to predict and evaluate vaginal mucosal toxicity induced by microbicides, and most importantly, they are unable to identify subtle or subclinical inflammatory reactions. This study was designed to develop a cost-effective in vivo model to evaluate microbicide safety in a preclinical study which can recapitulate the mucosal topical *** A murine model was employed with nonoxynol-9 （N-9） as the topical stimulant within the vagina. Different concentrations of N-9 （1%, 3%, and 4%） were topically applied to the vagina for five consecutive days. A panel of inflammatory cytokines including interleukine-2 （IL-2）, IL-4, IL-6, IL-17A, interferon-Y （IFN-Y）, tumor necrosis factor-α （TNF-α）, and immuno-regulatory IL-10 were assayed in vaginal lavage. Cytokines were quantified by using cytometric bead array （CBA） and reverse transcript （RT） real-time PCR. Histopathological evaluation of vaginal tissues was conducted on hematoxylin-eosin stained slides and scored with a semi-quantitative system according to the severity of epithelial disruption, leucocyte infiltration, edema, and vascular injection. The association between the cytokines and histopathological scores was assessed by linear regression *** All three concentrations of N-9 induced inflammatory cytokine production. The 4% N-9 application resulted in a consistent production of cytokines in a time-dependent manner. The cytokines reached peak expression on day three with the exception of IL-4 which reached its peak on day one. Histopathological examination of 4% N-9 treated cervicovaginal tissues on day three showed intensive damage in four mice （sores： 10-13） and moderate damage in one

关键词： hunman immunodeficiency virus 1 microbicide inflammatory cytokine murine model nonoxynol-9

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