MHC-BOUND, SIV-DERIVED, CTL AND HTL EPITOPES

在3000多万艾滋病毒感染者中，几乎没有比生产有效的艾滋病毒疫苗更紧迫的生物医学优先事项了。鉴于细胞毒性T淋巴细胞(CTL)和辅助T淋巴细胞(HTL)在控制病毒复制中的重要作用，这种疫苗刺激这些细胞反应是至关重要的。不幸的是，很少有MHC定义的恒河猴可用于疫苗研究。使用体外受精(IVF)和显微操作等辅助生殖技术，结合选定的胚胎移植，将产生完全由MHC定义的猕猴，表达多种MHC I和II类分子，其中存在SIV肽、四聚体和ELISPOT分析。在相关的病毒挑战模型中，MHC定义的和基因上相同的MHC定义的动物的效用将是巨大的，并对我们评估疫苗效力的能力产生巨大影响。我们已经确定了五对有多个后代的恒河猴。这五对动物将作为我们试管受精计划的卵子和精子捐赠者，并将构成我们MHC定义的育种计划的创始动物。为了确定体外受精捐赠者的MHC I和II类分子，我们将用SIV感染这五对猕猴的后代，然后确定对病毒的细胞免疫反应。这将使我们能够合成MHC I类和II类四聚体。具体目标1：我们将确定至少15个CTL表位及其限制四聚体合成的MHC I类等位基因。在特定目标2：我们将确定至少10个HTL表位及其限制四聚体合成的MHC II类等位基因。我们计划开发一个创新的原型动物生产项目，这将是地区灵长类中心未来的核心。利用辅助生殖技术生产的MHC定义的动物将补充猕猴四聚体和ELISPOT技术的最新发展，并为艾滋病疫苗研究提供宝贵的动物。新的MHC/多肽组合的定义和MHC定义的动物的生产一直是巴尔的摩AVRC委员会的优先事项(见巴尔的摩博士的支持信；附录)。

With more than 30 million HIV-infected individuals, there can be few other more pressing biomedical priorities than to produce an effective vaccine for HIV. Given the important role that cytotoxic T lymphocytes (CTLs) and helper T lymphocytes (HTLs) play in controlling viral replication, it is critical that this vaccine stimulate these cellular responses. Unfortunately, there are few MHC-defined rhesus macaques available for vaccine research. The use of assisted reproductive technologies such as in vitro fertilization (IVF) and micromanipulation in conjunction with selected embryo transfer will result in the production of completely MHC-defined macaques, expressing multiple MHC class I and II molecules for which SIV peptides, tetramers and ELISPOT assays exist. The utility of MHC-defined and genetically- identical MHC-defined animals in a relevant viral challenge model would be enormous and have tremendous impact on our ability to assess vaccine efficacy. We have identified five pairs of rhesus macaques that have multiple offspring. These five pairs of animals will serve as egg and sperm donors for our IVF program and will constitute the founding animals for our MHC-defined breeding program. To define the MHC class I and II molecules of the IVF donors, we will infect the offspring of these five pairs of macaques with SIV and then define the cellular immune response to the virus. This will enable us to synthesize both MHC class I and II tetramers. In Specific Aim 1: We will identify at least 15 CTL epitopes and their restricting MHC class I alleles for tetramer synthesis. In Specific Aim 2: We will identify at least 10 HTL epitopes and their restricting MHC class II alleles for tetramer synthesis. We plan to develop an innovative, prototypic animal production program that will be central to the future of the Regional Primate Centers. MHC-defined animals produced using assisted reproductive technologies will complement recent developments in tetramer and ELISPOT technologies in the rhesus macaque and provide invaluable animals for AIDS vaccine research. Definition of new MHC/peptide combinations and the production of MHC-defined animals has been a priority of the Baltimore AVRC Committee (see letter of support from Dr. Baltimore; Appendix).