OPTIMIZED HIV -SPECIFIC CTL ANTIGENS FOR VACCINE DESIGN

描述：(改编自申请者摘要)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的成功全球干预措施 将需要一种有效的疫苗。自诱导细胞毒作用以来 T淋巴细胞(CTL)反应现在被认为是一个重要的组成部分 有效的HIV-1疫苗，我们开发HIV疫苗的方法是基于 刺激CD8+CTL反应比自然感染更有效。 这将通过使用基于混合的合成组合来实现 文库(SCL)，以确定将是有效的优化的多肽配体 作为免疫原刺激T细胞介导的抗HIV免疫反应 感染。基于混合的SCL方法允许快速识别 从大量单个化合物中提取高活性化合物。在……里面 特别是，当以位置扫描(PS)格式生成时，关键氨基 活性多肽序列(S)的酸(S)可直接从 对图书馆进行初步筛选。CTL识别加工过的病毒多肽 通常长度为8到11个氨基酸，它们以分子的形式出现 与MHC-I类和β2-微球蛋白的复合体。非肽类化合物的PS-SCLS研究 十肽(具有羧酸或羧胺C-末端，以及 乙酰化或非乙酰化N-末端氨基)因此将用于 建议进行的研究。将对每种混合物进行筛选，以确定它们是否具有 CTL克隆具有刺激细胞因子产生和/或细胞溶解活性 免疫优势表位GAG、逆转录酶和 NEF蛋白。遵循去卷积过程以识别表位模拟物 从文库中，CTL对已识别的激动剂的反应将是 它们的免疫原性将通过体外实验进行评估 刺激性实验。因此，受冲击的细胞的免疫反应性 免疫原生肽和/或经鉴定的模拟物将被 通过细胞因子释放或铬释放试验测量。此外， 将研究最有效的识别多肽的交叉识别。 使用一组人类白细胞抗原A2阳性患者。虽然还不适用于艾滋病毒 研究表明，PS-SCL方法的成功已被报道为 多肽的鉴定比多肽的鉴定有效几个数量级 天然多肽配体刺激自身反应性克隆型群体 CD4+和肿瘤特异性或同种异体反应的CD8+T细胞。

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.