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

最近在申请人的实验室进行的研究 允许分子克隆和编码的部分特征 在来自非洲和北方的主要HIV-1亚型A，B，C和D中 美国的地方。已经确定了两个障碍， 拟议使用gp120克隆来开发全球 有效的亚单位疫苗。其中一个缺点是假定 菌株变异对抗原特异性和免疫原性的影响 Gp120的中和效果。全球血吸虫病血清阳性率 A、B、C和D亚型病毒尚未进行系统检查 举止。第二个问题是需要开发一种疫苗 可确保长期体内释放的制剂 免疫原性gp120多肽，并诱导终生杀菌免疫。 为了解决这些重要问题，可用的gp120克隆将 用于(A)确定艾滋病毒-1的相对血清流行率 A、B、C和D亚型；(B)优化技术，以长期 用血清流行病毒A、B、C和D亚型进行免疫； 和(C)评估特定类型或广泛的中和效力 在体外和体内的血清流行亚型如下 实验：一、确定和比较全球血吸虫病流行情况 编码于HIV-1 A、B、C和D亚型的gpl20。 要测试以下可能性，请执行以下操作： A.用来源于 血清流行的A、B、C和D亚型将在体内实现长期的 免疫原肽的递送 B.同时接种小鼠的ENV.GP120克隆 血清流行的A、B、C和D亚型引起广泛的反应 抗体和CTL中和具有可比性的不同分离株 效率。 C.接种ENV.GP120质粒克隆可产生抗体 以及中和HIV-1对CD4的感染性的CTL反应 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.