VAGINAL MICROBICIDES FOR PREVENTION OF HIV TRANSMISSION

产品说明：（改编自申请人的摘要）决定病毒如何通过生殖器上皮到达下层免疫细胞的机制知之甚少。因此，有必要建立一个模型来研究HIV-1生殖器感染事件以及评价杀微生物剂抑制HIV-1传播。在非人灵长类动物的研究提供了深入了解粘膜在SIV感染的作用，然而，使用不同的逆转录病毒和随后的疾病过程中，伦理问题和畜牧业的高成本，需要在体外试验，更好地复制人类生殖道环境的发展。使用小动物模型（小鼠和兔子）进行杀微生物剂测试存在类似的问题，因为这些动物不能感染HIV-1。本申请中提出的研究利用了使用原代宫颈上皮和基质免疫细胞的体外模型。将使用transwell系统中的原代上皮细胞和夹在扩散室中的宫颈上皮片评价杀微生物剂毒性。还将检查杀微生物剂抑制培养的粘膜免疫细胞中HIV-1感染和复制的能力。最后，宫颈移植模型将用于确定杀微生物剂在免疫激活或粘膜病原体降解上皮屏障期间抑制HIV-1传播到局部细胞的能力（T. pallidum和T.迷走神经）。拟议中的实验将为杀微生物剂的开发提供有价值的见解，并可能满足小动物试验的需要。

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.