PRODUCTION OF ANTI-HIV-1 THERAPEUTIC VACCINE

(1)项目目标： - 鉴定HIV-1亚单位， 中和抗体 - 为了鉴定将增加免疫原性的载体， HIV-1亚基，并将能够回忆起抗HIV B记忆细胞， 预先存在免疫缺陷的患者。 - 为了确定一个携带者，这将增加TH 1/TH 2比率的感染， 并将有利于产生细胞反应，包括 细胞毒性细胞（CTL）。 (2)实验方法： - 革兰氏阴性牛种布氏杆菌（Brucellabortus，Ba）及其细胞内脂多糖（LPS） 壁（Ba-LPS）作为灭活HIV-1或 病毒粒子、gp 120（SF 2）糖蛋白或衍生自V3环的肽 HIV-1（MN）env.将不同的偶联物用于免疫小鼠 都有不同程度的T细胞缺陷 - 使用来自正常人以及HIV-1感染者的PBL的体外研究 评估患者对Ba 和Ba-LPS。使用PCR和生物测定。 (3)主要发现： - Ba与含有B细胞表位和CTL表位的肽缀合 （N3 v3），产生中和抗体和细胞毒性T细胞 能够杀死感染艾滋病毒的目标 保留CD 4缺失小鼠 它们产生抗HIV中和Ab和CTL的能力， 用Ba-N3 V3缀合物免疫。 - 从安全的角度来看，Ba或其LPS对人的毒性要小得多。 动物比E.大肠杆菌衍生的LPS。 - Ba和Ba-LPS可直接激活纯化的人血浆， CD 4阳性TH 1细胞，以及在较小程度上，CD 8阳性细胞，如 通过淋巴因子IL 2和IFN-γ的诱导来判断。 来自HIV-1的PBL 感染者也有反应。 Ba和Ba-LPS还可以 激活人单核细胞的IL 12分泌。 出版物：（1）B。放大图片作者：J. Blackburn，J. Manischewitz和H. Golding 1955. J. Virology 69：3299-3307. (2)M. Zaitseva，H. Golding，M. Brtts，A. Yamauchi，E.布卢姆湖巴特勒湖 斯蒂文和B。戈尔丁1995年感染。和豁免权。63：2720-2728

(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 cytotoxic 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) B. Golding, J. Inman, P. Highet,, R. Blackburn, J. Manischewitz and H. Golding 1955. J. Virology 69:3299-3307. (2) M. Zaitseva, H. Golding, M. Brtts, A. Yamauchi, E. Bloom, L. Butler, L. Stevan, and B. Golding. 1995 Infec. and Immunity. 63: 2720-2728