NATURAL HIV-1 PEPTIDES AND CD8+ CYTOTOXIC T-CELLS

通过破坏病毒感染的细胞，CD8+细胞毒性T淋巴细胞（CTL） 有助于终止病毒感染， 在抵抗许多病毒的免疫防御中起重要作用。 虽然CTL 在人类感染的个体中发现了大量的 免疫缺陷病毒1型（HIV-1），并可能有助于降低病毒 在这些人的负担，这些细胞不能成功地消除 HIV-1感染。 为了帮助理解为什么这些细胞 有效的HIV-1感染，并帮助奠定基础， 更合理地开发疫苗， 生产和这些细胞的有效性，我们将确定和 分析HIV-1中HIV-1来源的天然肽 被感染的细胞 分析的肽将主要是那些 与I类MHC类蛋白结合被CD8 + 来自HIV-1感染个体的CTL克隆或系。 我们还将 分析与I类MHC相关的最丰富的HIV-1肽 感染细胞的蛋白质。 所有这些肽将被 根据它们的a）氨基酸序列，B）丰度 作为I类MHC蛋白的加合物，所述I类MHC蛋白分离自 产生HIV-1，和c）它们的亲和力（内在平衡 结合常数）。 为了实现这些目标，我们 将使用特别构建的稳定的培养细胞系， 每个细胞都能产生HIV病毒并表达I类MHC蛋白 选择代表人类中最常见的一种 并限制人细胞毒性T细胞对HIV-1肽的识别 细胞克隆 这些特别构建的HIV-1生产细胞系 和抗原加工突变细胞（如T2）， 外源性合成肽将被用作刺激细胞 从HIV-1感染者中诱导新的CD8 + CTL细胞系和克隆， 合适HLA类型（例如HLA-H2）的个体;细胞溶解性 将评估新克隆的有效性， 它们识别的（与MHC-1相关）也将被分析为 介绍了

By destroying virus-infected cells, CD8+ cytotoxic T lymphocytes (CTL) contribute to the termination of viral infections and play an important role in immune defenses against many viruses. Although CTL are found in substantial numbers in individuals infected with human immunodeficiency virus type 1 (HIV-1) and may help lower the viral burden in these individuals, these cells do not succeed in eliminating HIV-1 infection. To help understand why these cells are not more effective in HIV-1 infections and to help lay the groundwork for a more rational development of vaccines aimed at enhancing the production and effectiveness of these cells, we will identify and analyze the naturally occurring peptides of HIV-1 origin in HIV-1 infected cells. The peptides analyzed will be principally those that are recognized in association with class I MHC class proteins by CD8+ CTL clones or lines from HIV-1 infected individuals. We will also analyze the most abundant HIV-1 peptides associated with class I MHC proteins from infected cells. All of these peptides will be characterized in terms of their a) amino acid sequence, b) abundance as adducts of class I MHC proteins isolated from cells that are producing HIV-1, and c) their affinity (intrinsic equilibrium association constant) for MHC-I proteins. To achieve these goals we will use specially constructed stable, cultured cell lines in which every cell produces HIV virus and expresses a class I MHC protein chosen to represent one of the more common ones in human populations and that restrict recognition of HIV-1 peptides by human cytotoxic T cell clones. These specially constructed HIV-1 producing cell lines and antigen-processing mutant cells (such as T2) that can be loaded exogenously with synthetic peptides will be used as stimulator cells to elicit new CD8+ CTL cell lines and clones from HIV-1 infected individuals of appropriate HLA type (e.g. HLA-H2); the cytolytic effectiveness of the new clones will be evaluated and the peptides they recognize (in association with MHC-1) will also be analyzed as described.