OPTIMIZED HIV -SPECIFIC CTL ANTIGENS FOR VACCINE DESIGN

用于疫苗设计的优化 HIV 特异性 CTL 抗原

• 批准号: 6511432
• 负责人: SYLVIE E BLONDELLE
• 金额: $ 27万
• 依托单位: TORREY PINES INST FOR MOLECULAR STUDIES
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6511432
• 关键词: AIDS vaccines; MHC class I antigen; T cell receptor; biomimetics; cell mediated lymphocytolysis test; combinatorial chemistry; cytokine; cytotoxic T lymphocyte; human immunodeficiency virus 1; human tissue; molecular cloning; peptide chemical synthesis; peptide library; peptides; vaccine development

DESCRIPTION: (Adapted from Applicant's Abstract) Despite the great advances in antiviral therapy for human immunodeficiency virus (HIV) infection, a successful global intervention for prevention and treatment of HIV infection will require an effective vaccine. Since the induction of cytotoxic T-lymphocyte (CTL) responses is now believed to be an important component in an effective HIV-1 vaccine, our approach to develop an HIV vaccine is based on the stimulation of CD8+ CTL response more efficiently than the natural infection. This will be accomplished by using mixture-based synthetic combinatorial libraries (SCLs) to identify optimized peptide ligands that would be effective as immunogens in stimulating T-cell mediated immune responses against HIV infection. The mixture-based SCL approach allows the rapid identification of highly active compounds from large pools of individual compounds. In particular, when generated in a positional scanning (PS) format, the key amino acid(s) of the active peptide sequence(s) can be determined directly from the initial screening of the library. CTLs recognize processed viral peptides generally 8 to 11 amino acids in length, which are presented as a molecular complex with MHC class I and Beta2-microglobulin. PS-SCLs of nonapeptides and decapeptides (having a carboxylic acid or carboxamide C-terminus, and an acetylated or non-acetylated N-terminal amino group) will therefore be used for the proposed studies. Each mixture will be screened for their ability to stimulate cytokine production and/or cytolytic activity by CTL clones having specificity for immunodominant epitopes of the Gag, reverse transcriptase, and Nef proteins. Following the deconvolution processes to identify epitope mimics from the libraries, the CTL reactivities to the identified agonists will be determined, and their immunogenicity will be assessed by performing in vitro stimulation experiments. Thus, the immunologic reactivity of cells pulsed with peptides to the immunizing native peptide and/or identified mimics will be measured by cytokine release or chromium release assays. Furthermore, the cross-recognition of the most potent identified peptides will be investigated using a cohort of HLA-A2-positive patients. Although not yet applied to HIV research, the success of the PS-SCL approach has been reported for the identification of peptides being several order of magnitude more effective than native peptide ligands to stimulate clonotypic populations of autoreactive CD4+, and tumor-specific or alloreactive CD8+ T-cells.

描述：（改编自申请人的摘要）尽管在 人类免疫缺陷病毒（HIV）感染的抗病毒治疗， 预防和治疗艾滋病毒感染全球干预措施取得成功 需要有效的疫苗由于诱导细胞毒性 现在认为T淋巴细胞（CTL）应答是免疫应答的重要组成部分。 有效的HIV-1疫苗，我们开发HIV疫苗的方法是基于 比自然感染更有效地刺激CD 8 + CTL应答。 这将通过使用基于混合物的合成组合 库（SCL），以确定优化的肽配体，将是有效的 作为免疫原刺激T细胞介导的抗HIV免疫应答 感染基于混合的SCL方法可以快速识别 高活性化合物从单个化合物的大池。在 特别地，当以位置扫描（PS）格式生成时，关键氨基酸 活性肽序列的氨基酸可以直接从氨基酸序列中确定。 对文库进行初步筛选。CTL识别加工的病毒肽 通常长度为8至11个氨基酸，其以分子形式存在 与MHC I类和β 2-微球蛋白的复合物。九肽的PS-SCL和 十肽（具有羧酸或羧酰胺C-末端，和 乙酰化或非乙酰化的N-末端氨基）将因此用于 建议的研究。将筛选每种混合物的能力， 通过CTL克隆刺激细胞因子产生和/或细胞溶解活性，所述CTL克隆具有 对Gag的免疫显性表位、逆转录酶和 Nef蛋白。遵循去卷积过程以鉴定表位模拟物 从文库中，CTL对所鉴定的激动剂的反应性将被 测定，并通过体外试验评估其免疫原性 刺激实验因此，细胞的免疫反应性脉冲与 肽与免疫天然肽和/或鉴定的模拟物的结合将被 通过细胞因子释放或铬释放测定来测量。而且 将研究最有效的已鉴定肽的交叉识别 使用HLA-A2阳性患者队列。虽然尚未应用于艾滋病毒 研究，PS-SCL方法的成功已被报道为 肽鉴定比肽鉴定有效几个数量级 天然肽配体刺激自身反应性的克隆型群体 CD 4+和肿瘤特异性或同种异体反应性CD 8 + T细胞。