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

作　　者：LI Liang-zhu YANG Yu YUAN Song-hua WAN Yan-min QIU Chao FENG Yan-ling XU Jian-qing ZHANG Xiao-yan 

机构地区：[1]Research Center of Shanghai Public Health Clinical Center,Institutes of Biomedical Sciences, Fudan University, Shanghai 201508, China [2]Key Laboratory of Medical Molecular Virology & Institute of Medical Microbiology, Shanghai Medical College of Fudan University, Shanghai 200433, China [3]State Key Laboratory for Infectious Disease Prevention andControl, Chinese Center for Disease Control and Prevention,Beijing 100050, China

出　　处：《Chinese Medical Journal》2010年第23期3381-3388,共8页中华医学杂志（英文版）

摘　　要：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 reaction.Methods 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 analysis.Results 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 mouse �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 reaction.Methods 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 analysis.Results 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 mouse �

关 键 词：hunman immunodeficiency virus 1 MICROBICIDE inflammatory cytokine murine model NONOXYNOL-9 

分 类 号：R686[医药卫生—骨科学]