PRODUCTION OF ANTI-HIV-1 THERAPEUTIC VACCINE

(1)项目目标：确定将增加 HIV-1亚基的免疫原性，并将能够召回抗HIV 既往免疫缺陷患者的B记忆细胞。至 确定可增加感染者TH1/TH2比率的携带者 个体，并将有利于产生细胞反应，包括 细胞毒细胞(CTL)。(2)实验方法：革兰氏阴性 流产布鲁氏菌(BA)及其细胞壁来源的内毒素(BA-LPS)分别为 作为灭活的HIV-1病毒粒子gp120(SF2)的携带者检测 糖蛋白，或源自HIV-1(MN)env的V3环的多肽。这个 用不同的结合物免疫不同程度的小鼠 T细胞缺乏症。四只恒河猴也接种了疫苗 BA-V3结合物。体液免疫和细胞毒性免疫反应都是 测量包括全身和粘膜抗体反应。生物学上的 合胞体抑制试验检测相关抗体。在……里面 人T细胞和正常人外周血单核细胞的体外研究 以及对HIV-1感染患者的淋巴因子进行评估 用聚合酶链式反应和生物测定法测定对巴和巴-内毒素的反应产物。 (3)主要发现：BA与含有B细胞表位的多肽结合 和CTL表位(N3v3)，产生中和抗体和 能够杀死HIV感染目标的细胞毒T细胞。CD-4耗尽 小鼠保留产生抗HIV中和抗体和CTL的能力 用巴-N3V3结合物免疫后。从安全点上看 认为，巴或其内毒素对动物的毒性远低于大肠杆菌来源 LP。BA和BA-内毒素可直接激活纯化的人 CD4阳性的TH1细胞，程度较轻的CD8阳性细胞，AS 通过对淋巴因子IL2和干扰素-γ的诱导来判断。来自HIV-1的PBL 受感染的人也有反应。BA和BA-内毒素也可以 激活人单核细胞分泌IL-12。生产恒河猴 血清和粘膜中的高效价HIV-1中和抗体 表面。

(1) Goals of Project: 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. Four Rhesus macaques were also vaccinated with a Ba-V3 conjugate. Both humoral and cytotxic immune responses were measured including systemic and mucosal antibody responses. Biologically relevant antibodies were measured in syncytia inhibition assays. In vitro studies with human T cells and elutriated monocytes from normal as well as HIV-1 infected patients were assessed for their lymphokine production in response to Ba and Ba-LPS by PCR and biological assays. (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. Rhesus macacques produced high titer HIV-1-neutralizing antibodies in the serum and mucosal surfaces.