DESIGN OF MINI-GENES AS DNA VACCINES FOR HIV THERAPY

疫苗接种对预防和控制 几个世纪以来的传染病减毒活或死疫苗注射 病毒，产生特异性体液和细胞免疫， 重要的是，诱导免疫记忆。这保护了一个人 当受到致病病毒的攻击时。最近，行政 产生免疫力的单一重组病毒蛋白质已经遇到了 有限的成功。 目前还没有防治人体免疫缺陷病毒（HIV-1）的疫苗。这 是因为病毒感染并破坏了T淋巴细胞和巨噬细胞 它们是特异性免疫的关键调节剂。越来越多的证据 表明病毒的蛋白质产物本身对人类有害， 不仅是周围的细胞，还有大脑中的细胞。比如说 病毒的包膜蛋白，称为gp 12 O，即使在没有 感染，可以介导大脑中的毒性作用， 艾滋病痴呆综合征（ADC）。 注射编码完整病毒蛋白的DNA是一种令人兴奋的， 成功的疫苗接种新方法。在这份提案中，我们将判断 设计小基因DNA载体对CTL产生的影响 编码最小病毒表位，有和没有信号序列， 辅助性T细胞表位的存在和不存在，以及评估 细胞因子IL-2和IL- 12对CTL诱导的共刺激作用。 这种方法确保动物不会暴露于整个病毒， 蛋白质及其潜在的有害作用。该模型系统将 为制定发展总体战略奠定基础 产生含有多个病毒表位的载体， 在人群中产生免疫力。 在这个建议中，我们计划了四个具体目标：（1）设计载体， DNA疫苗接种仅编码所需的动物肽序列 在小鼠中产生gp 120特异性细胞毒性T细胞（CTL）;（2）检测 CD 4 + T辅助细胞在诱导这些gp 120- 特异性CTL;（3）确定是否可以增强CTL生成 通过共注射细胞因子表达载体，IL-2或IL- 12， 与编码gp 120 CTL表位的载体一起;（4）在 与L博士合作Mucke（Scripps），以评估 gp 120特异性CTL在转基因小鼠脑中表达 gp 120在星形胶质细胞中的特异性表达。

Vaccination has had a profound effect on the prevention and control of infectious disease for centuries. Injection of attenuated live or dead viruses, generates specific humoral and cellular immunity, and importantly, induces immunological memory. This protects an individual when challenged later by the pathogenic virus. Recently, administration of single recombinant viral proteins to generate immunity has met with limited success. No vaccine exists to combat human immunodeficiency virus (HIV-1). This is because the virus infects and cripples T lymphocytes and macrophages which are critical regulators of specific immunity. Increasing evidence suggests that protein products of the virus are themselves harmful to cells not only in the periphery, but also in the brain. For example, the envelope protein of the virus, called gp12O, even in the absence of infection, can mediate toxic effects in the brain which resemble those observed in AIDS dementia complex (ADC). Injection of DNA encoding complete viral proteins is an exciting and successful new method for vaccination. In this proposal, we will judge the efficacy for CTL generation of designing mini-gene DNA vectors encoding minimal viral epitopes, with and without signal sequences, in the presence and absence of T helper epitopes as well as assess the costimulatory effects of the cytokines IL-2 and IL- 12, on CTL induction. This approach ensures that animals are not exposed to entire viral proteins with their potential deleterious effects. This model system will lay tide foundation for the development of a general strategy for creating vectors containing multiple viral epitopes which can be used to generate immunity in human populations. In this proposal, we plan four Specific Aims: (1) to design vectors for DNA vaccination encoding only animal peptide sequences required to generate gp120-specific cytotoxic T cells (CTLs) in mice; (2) to examine the role which CD4+ T helper cells play in the induction of these gp120- specific CTLs; (3) to determine whether CTL generation can be enhanced by coinjection of a cytokine expression vector, either IL-2 or IL- 12, together with the vector encoding the gp120 CTL epitope; (4) in collaboration with Dr. L. Mucke (Scripps), to assess the effect that gp120-specific CTLs have on the brains of transgenic mice expressing gp120 specifically in astrocytes.