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

通过破坏感染病毒的细胞，CD8+细胞毒性T淋巴细胞(CTL) 为终止病毒感染做出贡献，并发挥 在对多种病毒的免疫防御中发挥重要作用。虽然CTL 在感染人类的人身上发现了大量的 免疫缺陷病毒1型(HIV-1)，可能有助于降低病毒 这些个体的负担，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没有更多 在HIV-1感染中有效，并帮助为 更合理地开发疫苗，旨在提高 这些细胞的生产和有效性，我们将确定和 HIV-1中HIV-1来源的天然多肽的分析 被感染的细胞。分析的多肽将主要是那些 被CD8+识别为与I类MHC类蛋白相关 来自HIV-1感染者的CTL克隆或系。我们还将 分析与I类MHC相关的最丰富的HIV-1多肽 来自受感染细胞的蛋白质。所有这些多肽都将是 其特征在于a)氨基酸序列，b)丰度 作为从细胞中分离的I类MHC蛋白的加合物 产生HIV-1，以及c)它们的亲和力(内在平衡 MHC-I蛋白的结合常数)。为了实现这些目标，我们 将使用专门构建的稳定的、培养的细胞系 每个细胞都会产生HIV病毒，并表达一种I类MHC蛋白 被选来代表人类中最常见的一种 限制人细胞毒T细胞对HIV-1多肽的识别 细胞克隆。这些特殊构建的HIV-1产生细胞系 和抗原处理突变细胞(如T2)，可以加载 外源性合成多肽将被用作刺激细胞 从HIV-1感染者体内诱导新的CD8+CTL细胞株和克隆 适当的人类白细胞抗原类型的个体(例如，人类白细胞抗原-H2)；细胞溶解 将对新克隆的有效性进行评估，并将多肽 他们认识到(结合MHC-1)也将被分析为 描述。

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.