VAGINAL MICROBICIDES FOR PREVENTION OF HIV TRANSMISSION

描述：（改编自申请人摘要）关于病毒如何通过生殖上皮到达底层免疫细胞的机制尚不清楚。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估杀微生物剂以抑制HIV-1传播。对非人灵长类动物的研究已经深入了解了SIV感染期间粘膜的作用；然而，由于使用不同的逆转录病毒和随后的疾病过程，再加上伦理问题和畜牧业的高成本，有必要开发更好地复制人类生殖道环境的体外测定法。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HIV-1。在本应用中提出的研究利用了一种体外模型，该模型使用了原代宫颈上皮细胞和间质免疫细胞。杀微生物剂的毒性将在transwell系统中对原代上皮细胞进行评估，并将宫颈上皮片夹在扩散室中。在培养的粘膜免疫细胞中，杀微生物剂抑制HIV-1感染和复制的能力也将被检验。最后，宫颈外植体模型将用于确定杀微生物剂在粘膜病原体（T. pallidum和T. vaginalis）免疫激活或上皮屏障降解过程中抑制HIV-1向局部细胞传播的能力。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.