PRODUCTION OF ANTI-HIV-1 VACCINE

(1)项目目标：-确定艾滋病毒-1亚基将产生 广泛交叉中和抗体。-确定承运人将 提高HIV-1亚单位的免疫原性，并将能够 已有免疫的患者召回抗HIV B记忆细胞 缺乏症。-识别可增加TH1/TH2比率的载波 并将有利于产生细胞反应 包括细胞毒细胞(CTL)。(2)实验方法：--克 阴性流产布鲁氏菌(BA)和来源于细胞壁的内毒素(BA- 作为灭活的HIV-1病毒粒子gp120的携带者 (SF2)糖蛋白，或源自HIV-1(MN)V3环的多肽 环境用不同的结合物免疫不同类型的小鼠。 T细胞缺乏的程度。-正常PBL的体外研究 对人类和HIV-1感染患者进行了评估 巴和巴-内毒素对淋巴因子产生的影响。既有聚合酶链式反应又有 采用生物测定法。(3)主要发现：-BA与A 含有B细胞表位和CTL表位(N3v3)的肽，两者都产生 中和抗体和细胞毒性T细胞能够杀死HIV- 被感染的目标。CD4耗尽的小鼠保留了它们产生 巴-N3V3免疫后的抗HIV中和抗体和CTL 共轭。-从安全的角度来看，巴或它的脂多糖要少得多 对动物的毒性大于E.Coli来源的内毒素。-巴和巴-内毒素被发现 直接激活纯化的人CD4阳性TH1细胞，并 较小程度的CD8阳性细胞，通过诱导 淋巴因子IL-2和干扰素-γ。HIV-1感染者的PBL 反应也很灵敏。BA和BA-LPS还可通过以下途径激活IL12的分泌 人类单核细胞-出版：(1)H.Golding，P.D‘Souza，J.Bradac， B.Mathieson和P.Fast。“艾滋病毒-1的中和：技术和 催乳素疫苗临床试验分析试剂“艾滋病” 研究与人类逆转录病毒。1994年。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.