PRODUCTION OF ANTI-HIV-1 VACCINE

(1)项目目标：-确定HIV-1亚单位， 广泛交叉中和抗体。- 确定一个承运人， 增加HIV-1亚单位的免疫原性， 在预先存在免疫缺陷的患者中回忆抗HIV B记忆细胞 缺陷- 确定一种能够增加TH 1/TH 2比值的载体 并将有利于产生细胞反应 包括细胞增殖细胞（CTL）。(2)实验方法：-克 阴性流产布鲁氏菌（Ba）和来自其细胞壁的LPS（Ba- LPS）作为灭活的HIV-1病毒粒子、gp 120 (SF2)糖蛋白或衍生自HIV-1（MN）V3环的肽 env.使用不同的缀合物以不同的抗体免疫小鼠。 T细胞缺乏的程度。- 来自正常人的PBL的体外研究 评估人类以及HIV-1感染患者的 淋巴因子的产生响应Ba和Ba-LPS。pcr和 使用生物测定法。 (3)主要发现：- Ba与a 含有B细胞表位和CTL表位的肽（N3 v3），产生了 中和抗体和能够杀死HIV的细胞毒性T细胞- 被感染的目标 CD 4耗竭的小鼠保留了它们产生 用Ba-N3 V3免疫后的抗HIV中和Ab和CTL 共轭。从安全性角度考虑，Ba或其LPS要少得多 对动物的毒性比E.大肠杆菌衍生的LPS。 - 发现Ba和Ba-LPS 直接活化纯化的人CD 4阳性TH 1细胞， 程度较轻，CD 8阳性细胞，通过诱导 淋巴因子IL 2和IFN-γ。 来自HIV-1感染者的PBL是 也有反应。 Ba和Ba-LPS还可以通过以下方式激活IL 12分泌： 人类单核细胞发表论文：（1）H.戈尔丁，P. D 'Souza，J. Bradac， B。Mathieson，and P. Fast. “HIV-1的中和作用：技术和 艾滋病疫苗临床试验分析试剂 研究和人类逆转录病毒。1994. 10：633-643.

(1) Goals of Project: - To identify HIV-1 subunits which will generate widely cross neutralizing antibodies. - To identify a carrier which will increase the immunogenicity of the HIV-1 subunits, and will be able to recall anti-HIV B memory cells in patients with pre- existing immune deficiency. - To identify a carrier which would augment the TH1/TH2 ratio of infected individuals and will favor generation of cellular responses including cytotxic cells (CTL). (2) Experimental approaches: - The gram negative Brucella abortus (Ba), and LPS derived from its cell wall (Ba- LPS), were tested as carriers for either inactivated HIV-1 virions, gp120 (SF2) glycoprotein, or peptide derived from the V3-loop of HIV-1 (MN) env. The different conjugates were used to immunize mice with different degrees of T cell deficiency. -In vitro studies with PBL from normal human as well as HIV-1 infected patients were assessed for their lymphokine production in response to Ba and Ba-LPS. Both PCR and biological assays were used. (3) Major findings: - Ba conjugated to a peptide containing B-cell epitope and CTL epitope (N3v3), generated both neutralizing antibodies and cytotoxic T cells capable of killing HIV- infected targets. CD4 - depleted mice retained their ability to generate anti-HIV neutralizing Ab and CTL after immunization with the Ba-N3V3 conjugate.- From a safety point of view, Ba or its LPS are much less toxic to animals than E. Coli derived LPS. - Ba and Ba-LPS were found to directly activate purified human CD4-positive TH1 cells, and to a lesser degree, CD8-positive cells, as judged by induction of the lymphokines IL2 and IFN-gamma. PBL from HIV-1 infected individuals were also responsive. Ba and Ba-LPS can also activate IL12 secretion by human monocytes.- Publications: (1) H. Golding, P. D'Souza, J. Bradac, B. Mathieson, and P. Fast. "The neutralization of HIV-1: Technology and reagents for analysis of prohylactic vaccines clinical trials" AIDS Research and Human Retroviruses. 1994. 10: 633-643.