T CELL EPITOPES OF THE MALARIA CS PROTEINS

为了优化疟疾合成肽的免疫应答 疫苗、寄生虫来源的"通用" T细胞表位，即 可以结合大多数II类分子，将在 疟疾红细胞前期抗原及其特异性和效应物 确定含有这些表位的疫苗引起的功能。 在 在目前的资助下，我们建议研究我们认为的通用T细胞表位 用两种实验方法鉴定了。 一种是 直接或经验性的，并利用疟疾特异性人类T细胞克隆， 鉴定P.的aa 326 - 345中的潜在通用T细胞表位。 恶性疟原虫（NF 54）CS蛋白。 用该方法确定的抗原表位， 在体外与多个DR和DQ分子结合并引发抗体 不同遗传背景的小鼠的反应。 第二种方法我们 已经使用身份"理论"通用T细胞表位，通过分析 含有多个高亲和力HLA的区域的蛋白质序列 肽结合基序。 自提交原始申请以来，我们 已经获得了初步的结果，证明了使用基序的能力， 分析以鉴定P. TRAP/SSP 2蛋白。 混杂结合的分子基础 HLA-II类分子的免疫学定义和预测的P。 恶性疟原虫通用T细胞表位将使用体外肽来定义 结合测定。 含有疟疾通用T细胞的合成肽 将测试表位被子孢子免疫的细胞识别 志愿者和不同单倍型的鼠品系。 的新方法 构建含有多个寄生虫的复合多聚体肽- 衍生的T和B细胞表位，通过MAP技术或多肟合成 肽化学，也将探索。 三表位MAP或多肟 构建体，其含有恶性疟原虫CS或Trap通用T细胞 表位，与CS重复B细胞表位组合，将被测试 因为它们能够诱导疟疾特异性抗体和细胞介导的 不同遗传背景的小鼠品系的免疫应答。 的 CS和TRAP通用表位保护性疟疾类似物的研究 对抗子孢子攻击。 肽/MHC相互作用的相关性 在体外与体内功能免疫原性将验证使用 开发能够诱发疟疾疫苗的方法 保护水平的体液和细胞免疫的个人 不同的遗传背景。

In order to optimize the immune response to malaria synthetic peptide vaccines, parasite-derived "universal" T cell epitopes, i.e. epitopes that can bind to the majority of class II molecules, will be identified in malaria preerythrocytic stage antigens and the specificity and effector functions elicited by vaccines containing these epitomes determined. In the current grant, we propose to study universal T cell epitopes that we have identified using two experimental methods. The first method was direct, or empirical, and utilized malaria-specific human T cell clones to identify a potential universal T cell epitope in aa 326-345 of the P. falciparum (NF54) CS protein. The epitope defined by this method was shown to bind to multiple DR and DQ molecules in vitro and elicit antibody responses in mice of diverse genetic backgrounds. The second method we have used identities "theoretical" universal T cell epitopes by analyzing protein sequences for regions that contain multiples high affinity HLA peptide binding motifs. Since submission of our original application, we have obtained preliminary results demonstrating the ability to use motif analysis to identify three potential universal T cell epitopes in the P. falciparum TRAP/SSP2 protein. The molecular basis of promiscuous binding to HLA class II molecules of the empirically-defined and the predicted P. falciparum universal T cell epitopes will be defined using in vitro peptide binding assays. Synthetic peptides containing the malaria universal T cell epitopes will be tested for recognition by cells of sporozoite-immunized volunteers and murine strains of different haplotypes. New methods for the construction of complex multimeric peptide containing multiple parasite- derived T and B cell epitopes, synthesized by MAP technology or polyoxime peptide chemistry, will also be explored. Tri-epitope MAP or polyoxime constructs, containing the P. falciparum CS or Trap universal T cell eptiopes, incombination with the CS repeat B cell epitope, will be tested for their ability to induce malaria-specific antibody and cell-mediated immune responses in murine strains of diverse genetic backgrounds. The muring malaria analogues of CS and TRAP universal epitopes inprotectin against sporozoite challenge. The correlation for peptide/MHC interactions in vitro with function immunogenicity in vivo will validate the use of approaches for the development of malaria vaccines capable of eliciting protective levels of humoral and cellular immunity in individuals of diverse genetic backgrounds.