SALMONELLA-HIV CHIMERA AS VACCINE

沙门氏菌-HIV 嵌合体作为疫苗

• 批准号: 3141946
• 负责人: BRUCE A STOCKER
• 金额: $ 25万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-12-01 至 1993-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3141946
• 关键词: AIDS vaccines; Salmonella; Salmonella vaccines; T lymphocyte; antibody formation; chimeric proteins; drug screening /evaluation; flagellin; flagellum; genetic manipulation; guinea pigs; human immunodeficiency virus; laboratory mouse; leukocyte activation /transformation; live vaccine; molecular cloning; neutralizing antibody; synthetic nucleotide; virus antigen

The long-term objective is to construct Salmonella-based anti-HIV vaccine strains for trial in volunteers, consisting of an aromatic-dependent strain of S. typhi (needing p-aminobenzoate, not available in host tissues, therefore non-pathogenic) carrying genetic information such that its administration will cause humoral and cellular immune responses to both the bacterial carrier and to the HIV epitope encoded by the passenger DNA. The foreign DNA will be a short synthetic oligonucleotide specifying an amino acid sequence identified as an epitope of an HIV protein antigen able to cause production of virus-neutralizing antibody or other protective response. The oligo will be inserted in a cloned flagellin gene at a point such that the amino acid sequence it specifies is exposed at the flagellar surface, accessible to antibody and expected to be highly immunogenic. Another form of the vaccine will consist of flagella broken off from the live-vaccine bacteria, concentrated and purified for use as a product vaccine, to be given by injection. Specific Aim 1 is to test immune responses of mice and guinea pigs to parenteral or oral administration of a Salmonella live vaccine with flagella made of flagellin specified by a cloned gene with oligos determining short amino acid sequences from gp160 of HIV inserted at the EcoRV site, by use of already constructed or to be made derivatives of plasmid pLS408 placed in a flagellin-negative aroA live-vaccine strain of S. dublin. Aim 2 is development of a method for quantitation of flagellin and the investigation of what features of an amino acid insert determine continued production of flagella or the accumulation or secretion of flagellin without production of flagella; and of genetic methods of increasing amount of flagellin produced by a strain and the effect on immunogenicity, etc, of having two or more inserts, specifying the same or different epitopes, in tandem in a single flagellin molecule. Aim 3 is the construction of a replacement for a live-vaccine strain of S. typhi already tested in volunteers by one attenuated by two deletions in the aro (aromatic biosynthesis) pathway, instead of by an aroA deletion and a purA deletion, and investigation of effect of variation of parameters of dosage, presentation, etc, on efficacy of orally administered live vaccine.

长期目标是构建基于沙门氏菌的抗HIV疫苗 在志愿者身上试验的菌株，由一种芳香依赖菌株组成 伤寒沙门氏菌(需要对氨基苯甲酸盐，宿主组织中没有， 因此非致病的)携带遗传信息，使得其 给药会引起体液和细胞免疫反应 细菌载体和乘客DNA编码的HIV表位。这个 外来DNA将是一种短的合成寡核苷酸，指定一种氨基酸 被鉴定为HIV蛋白抗原表位的酸性序列 导致产生病毒中和抗体或其他保护性措施 回应。该寡核苷酸将被插入到克隆的鞭毛蛋白基因中。 这样它指定的氨基酸序列就暴露在鞭毛上 表面，可与抗体接触，预计具有高度的免疫原性。 另一种形式的疫苗将由从疫苗表面脱落的鞭毛组成 活疫苗细菌，浓缩和提纯后作为产品使用 疫苗，以注射方式给予。 具体目的1是测试小鼠和豚鼠对 注射或口服沙门氏菌活疫苗 由寡聚基因克隆的鞭毛蛋白制成的鞭毛 人类免疫缺陷病毒gp160的短氨基酸序列测定 EcoRV网站，通过使用已经建造或将制成的衍生品 将pLS408载体置于鞭毛蛋白阴性的AroA活疫苗株中 美国都柏林。 目标2是建立鞭毛蛋白的定量方法和 对氨基酸插入的哪些特征的研究继续进行 鞭毛的产生或鞭毛蛋白的积累或分泌 不产生鞭毛；以及增加数量的遗传方法 一株菌株产生的鞭毛蛋白及其对免疫原性等的影响 有两个或两个以上的插入，指定相同或不同的表位 在单个鞭毛分子中串联。 目标3是构建一种活疫苗株的替代品。 伤寒已经在志愿者中进行了测试，其中一人因两个缺失而变弱 ARO(芳香生物合成)途径，而不是通过ARO缺失和 Pura删除，并考察了参数变化的影响 关于口服活疫苗效力的剂量、陈述等。