VAGINAL MICROBICIDES FOR PREVENTION OF HIV TRANSMISSION

描述：(改编自申请人的摘要)确定病毒如何通过生殖器上皮到潜在免疫细胞的机制还知之甚少。因此，需要一种模型来研究艾滋病毒-1生殖器感染事件以及评价用来抑制艾滋病毒-1传播的杀微生物剂。对非人类灵长类动物的研究已经为SIV感染过程中粘膜的作用提供了洞察力；然而，使用不同的逆转录病毒和随后的疾病过程，加上伦理问题和高昂的畜牧业成本，有必要开发更好地复制人类生殖道环境的体外检测方法。使用小动物模型(小鼠和兔子)进行杀微生物剂测试也存在类似的问题，因为这些动物无法感染艾滋病毒-1。本申请中提出的研究利用了一种体外模型，该模型同时使用了原代宫颈上皮和间质免疫细胞。杀菌剂的毒性将通过Transwell系统中的原代上皮细胞和夹在扩散室中的宫颈上皮片进行评估。还将检测一种杀菌剂在培养的粘膜免疫细胞中抑制HIV-1感染和复制的能力。最后，宫颈移植模型将被用来确定在免疫激活或粘膜病原体(梅毒螺旋体和阴道毛滴虫)降解上皮屏障期间，杀微生物剂抑制HIV-1传播到局部细胞的能力。拟议的实验将为杀微生物剂的开发提供有价值的见解，并可能消除对小动物试验的需要。

DESCRIPTION: (Adapted from Applicant's Abstract) The mechanisms determining how virus is passed through the genital epithelium to underlying immune cells are poorly understood. Consequently, there is a need for a model to study the events of HIV-1 genital infection as well as evaluation of microbicides to inhibit HIV-1 transmission. Studies in nonhuman primates have provided insight into the role of the mucosa during SIV infection; however, use of a different retrovirus and the ensuing disease course, together with ethical issues and the high cost of animal husbandry, necessitate development of in vitro assays which better duplicate the human genital tract environment. Use of small animal models (mice and rabbits) for microbicide testing presents similar problems given the inability to infect these animals with HIV-1. The studies proposed in this application utilize an in vitro model that uses both primary cervical epithelial and stromal immune cells. Microbicide toxicity will be evaluated with primary epithelial cells in a transwell system and cervical epithelial sheets clamped into diffusion chambers. The ability of a microbicide to inhibit HIV-1 infection and replication in cultured mucosal immune cells will also be examined. Finally, the cervical explant model will be used to determine the ability of a microbicide to inhibit dissemination of HIV-1 to local cells during immune activation or degradation of the epithelial barrier by mucosal pathogens (T. pallidum and T. vaginalis). The proposed experiments will provide valuable insight into microbicide development and may obviate the need for small animal testing.