CHARACTERIZATION OF AFE INHIBITORS IN HUMAN CERVICAL/REC

研究项目I(评估人类粘膜外植体中的AFE抑制剂)的负责人将是Robin Shattock博士。该项目将设在英国伦敦的圣乔治医院医学院，Shattock博士和他的同事在那里开创了粘膜组织外植体(宫颈、阴道、直肠和阴茎)的培养。人类宫颈组织外植体模型已成功地用于收集有关可能涉及HIV-1性传播的生物过程的有用新信息。在手术后直接建立的宫颈和直肠组织外植体中存在的细胞，代表了在性传播期间首次接触艾滋病毒-1的免疫群体的表型(例如，共受体表达)、状态 激活和解剖位置。因此，沙托克博士使用的模型系统最直接地反映了在进入的病毒对人类的粘膜攻击过程中发生了什么。在任何基于机理的合理杀微生物剂设计方法中，它都是一个很有前途的工具。因此，我们将评估由研究支持核心提供的AFE抑制剂，这些抑制剂针对HIV-1与CD4、CLR或CCR的相互作用，或者影响迁移树突状细胞对病毒的摄取和传播。这些研究的目的是：评估AFE抑制剂组合对来自多个基因亚型的一组不同的艾滋病毒-1初级分离株的活性，包括来自艾滋病毒-1从男性传播到女性传播特别常见的国家的病毒；确定 在生理相关条件下(在精液或血液存在的情况下，pH为4-8)抑制物的活性；确定对人体组织和细胞的潜在毒性或炎症作用；确定不同的AFE抑制物配方是否削弱其抗病毒活性，或增加其毒性。正在考虑的具体问题是：AFE抑制剂及其组合是否影响宫颈和直肠植入物的粘膜HIV-1感染？哪些AFE抑制剂或组合在抑制局部HIV-1感染和粘膜组织病毒的迁移细胞传播方面最有效？AFE抑制剂、联合制剂和制剂在外植体模型中显示出可接受的组织相容性吗？哪种配方 在不增加HIV-1感染的情况下提供有效的药物渗透？这种研究，再加上我们对病毒-细胞进入过程和动物模型的全面了解，将有助于合理选择潜在的AFE抑制剂组合进行灵长类研究，然后进行人类临床试验。

The leader of Research Project I (assessment of AFE inhibitors in human mucosal explants) will be Robin Shattock, PhD. The project will be located at St George's Hospital Medical School, London, UK, where Dr. Shattock and colleagues have pioneered the culture of mucosal tissue explants (cervical, vaginal, rectal and penile). The human cervical tissue explant model has been successfully used to glean useful new information on the biological processes that may be involved in the sexual transmission of HIV--1. Cells present within cervical and rectal tissue explants, established directly after surgery, represent the immune population first exposed to HIV-1 during sexual transmission in terms of phenotype (e.g., co-receptor expression), state of activation and anatomical location. Thus the model system used by Dr. Shattock most directly reflects what happens during mucosal challenge of humans by an incoming virus. It is a promising tool in any mechanism-based approach to rational microbicide design. Thus we will evaluate AFE inhibitors, provided by the Research Support Core, that target HIV-1 interactions with CD4, CLR or CCR, or that impact on virus uptake and dissemination by migratory dendritic cells. The purpose of these studies is to: evaluate AFE inhibitor combinations for their activity against a diverse set of primary HIV-1 isolates from multiple genetic subtypes, including viruses from countries where the male-to-female spread of HIV-1 is particularly common; define the activity of inhibitors under physiologically relevant conditions (pH 4-8, in the presence of semen or blood); identify potential toxic or inflammatory effects in human tissues and cells; determine whether the formulation of different AFE inhibitors impairs their antiviral activity, or increases their toxicity. The specific questions being considered are: Do AFE inhibitors and combinations influence mucosal HIV-1 infection of cervical and rectal explants? Which AFE inhibitors or combinations are most effective at inhibiting both localized HIV-1 infection and migratory cell dissemination of virus from mucosal tissue? Do AFE inhibitors, combination and formulations show acceptable tissue compatibility in explant models? Which formulation characteristics provide effective drug penetration without enhancement of HIV-1 infection? This kind of study, together with our overall knowledge of the virus-cell entry processes and animal models, will facilitate the rational selection of potential AFE inhibitor combinations for primate studies, then human clinical trials.