DESIGN OF MINI-GENES AS DNA VACCINES FOR HIV THERAPY

接种疫苗对预防和控制流行性感冒产生了深远的影响 几个世纪以来的传染病。注射减毒活的或死的 病毒，产生特定的体液和细胞免疫，以及 重要的是，它能诱导免疫记忆。这保护了一个人 当稍后受到致病病毒的攻击时。最近，政府 产生免疫力的单一重组病毒蛋白遇到了 有限的成功。 目前还没有对抗人类免疫缺陷病毒(HIV-1)的疫苗。这 是因为病毒感染并破坏T淋巴细胞和巨噬细胞 它们是特定免疫力的关键调节器。越来越多的证据 表明病毒的蛋白质产品本身对人体有害 细胞不仅在外周，而且在大脑中。例如， 病毒的包膜蛋白，称为gp120，即使在没有 感染，可以调节大脑中的毒性效应，类似于 在艾滋病痴呆综合征(ADC)中观察。 注射编码完整病毒蛋白的DNA是一种令人兴奋的和 成功的疫苗接种新方法。在这份提案中，我们将评判 设计微基因DNA载体对CTL生成的影响 编码最小病毒表位，有和没有信号序列，在 T辅助表位的存在和缺失以及评估 细胞因子IL-2和IL-12对CTL诱导的协同刺激作用 这种方法确保了动物不会接触到整个病毒 蛋白质及其潜在的有害影响。这一模式系统将 为制定……总体战略奠定潮汐基础 创建包含多个病毒表位的载体，可用于 在人类群体中产生免疫力。 在这个提案中，我们计划了四个具体目标：(1)为 仅编码动物多肽序列的DNA疫苗 在小鼠体内产生gp120特异性细胞毒性T细胞(CTL)；(2)检测 CD4+T辅助细胞在诱导这些gp120- 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， 连同编码gp120 CTL表位的载体；(4)在 与L.Mucke博士(斯克里普斯)合作，评估 Gp120特异性CTL对表达gp120的转基因小鼠大脑的影响 Gp120特异性存在于星形胶质细胞中。

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.