A8: GLOBAL HIV VIRAL SUBTYPES & VACCINE DVMT: CLONING OF GP120 FROM AFRICA

申请人实验室最近进行的研究表明， 允许分子克隆和部分表征编码 在非洲和北非的主要HIV-1亚型A、B、C和D中， 美国地方。 在这方面已查明两个障碍， 建议使用gp120克隆开发全球性的 有效亚单位疫苗。 其中一个缺点是假定 菌株变异对抗原特异性的影响， GP120的中和效力。 全球艾滋病毒/艾滋病血清阳性率 病毒亚型A、B、C和D尚未进行系统的研究。 方式 第二个担忧是需要开发疫苗 将确保药物的长期体内释放的制剂 免疫原性GP 120肽并引发终身灭菌免疫。 为了解决这些重要问题，可用的gp120克隆将 用于（a）确定HIV-1的相对血清流行率 亚型A、B、C和D;（B）优化用于长期治疗的技术 用血清流行病毒亚型A、B、C和D免疫; 和（c）评估所述化合物的类型特异性或广泛中和功效， 血清流行亚型，在体外和体内，在以下 一、确定和比较全球血清流行率， 在HIV-1亚型A、B、C和D中编码的gp120。 二.为了检验以下可能性： a.用来源于大肠杆菌的ENV.GPI20质粒克隆接种小鼠。 血清阳性亚型A、B、C和D。将在体内实现长期的 免疫原性肽的递送 B.用来自小鼠的ENV.GP120克隆同时接种小鼠。 血清流行亚型A、B、C和D显示广泛反应性 中和不同分离株的抗体和CTL， 效率 C.用ENV.GP120质粒克隆进行接种以产生抗体 中和HIV-1对CD4的感染性的CTL反应性 在SCID小鼠或灵长类动物模型中的细胞。

Studies recently conducted in the applicant's laboratory have permitted the molecular cloning and partial characterization encoded in the major HIV-1 subtypes A, B, C, and D from the African and North American localities. Two obstacles have been identified in the proposed use of the gp120 clones for the development of a globally efficacious subunit vaccine. One of the drawbacks is the presumed impact of strain variation on the antigenic specificity and neutralizing efficacy of gp120. The global seroprevalence of the virus subtypes A, B, C, and D has not been examined in a systematic manner. The second concern is the need to develop a vaccine formulation that would assure long-term in vivo release of the immunogenic gp120 peptides and elicit life-long sterilizing immunity. To address these important concerns, the available gp120 clones will be used to (a) determine the relative sero-prevalence of the HIV-1 subtypes A, B, C, and D; (b) optimize the technique for long-term immunization with the sero-prevalent virus subtypes A, B, C, and D; and (c) assess the type-specific or broad neutralizing efficacy of the sero-prevalent subtypes, both in vitro and in vivo in the following experiments: I.To determine and compare the global sero-prevalence of gpl20 encoded in the HIV-1 subtypes A, B, C, and D. Il.To test the possibilities that: a. Inoculation of mice with ENV.GPI2O plasmid clones derived from the seroprevalent subtypes A, B, C, and D. would achieve long-term in vivo delivery of immunogenic peptides b. Concomitant inoculation of mice with the ENV.GP120 clones from the seroprevalent subtypes A, B, C, and D elicits broadly reactive antibodies and CTL that neutralize divergent isolates at comparable efficiency. c. Inoculation with the ENV.GP120 plasmid clones elicits antibodies and CTL reactivity that neutralize the infectivity of HIV-1 for CD4 cells in the SCID mouse or primate model.