T CELL EPITOPES OF THE MALARIA CS PROTEINS

为了优化疟疾合成肽的免疫应答 疫苗，寄生虫衍生的“通用”T细胞表位，即 可以与大多数II类分子结合，将在 疟疾红细胞前期抗原及其特异性和效应物 确定了含有这些表位的疫苗所激发的功能。在……里面 目前的拨款，我们建议研究通用T细胞表位，我们 已经用两种实验方法进行了鉴定。第一种方法是 直接的，或经验性的，并利用疟疾特异性人类T细胞克隆来 鉴定P.A.AA326-345中可能的通用T细胞表位 恶性疟原虫(NF54)CS蛋白。用这种方法定义的表位显示了 体外与多个DR和DQ分子结合并诱导抗体 不同遗传背景的小鼠的反应。第二种方法是我们 通过分析使用了相同的“理论”通用T细胞表位 含有多个高亲和力人类白细胞抗原区域的蛋白质序列 多肽结合模体。由于我们提交了最初的申请，我们 已经获得了初步结果，证明了使用Motif的能力 分析鉴定P. 恶性疟原虫TRAP/SSP2蛋白。混杂结合的分子基础 对经验性定义的和预测的P. 恶性疟原虫通用T细胞表位将用体外多肽确定 有约束力的分析。含有疟疾通用T细胞的合成肽 将测试抗原表位以供免疫的子孢子细胞识别 志愿者和不同单倍型的小鼠品系。可持续发展的新方法 含多种寄生虫的复合多聚肽的构建 衍生的T和B细胞表位，通过MAP技术或多肟合成 多肽化学，也将进行探索。三表位图或多肟 含有恶性疟原虫CS或Trap通用T细胞的载体 将测试与CS重复B细胞表位结合的表位 诱导疟疾特异性抗体和细胞介导性抗体的能力 不同遗传背景的小鼠品系的免疫反应。这个 疟疾CS和TRAP通用表位类似物的保护性研究 抗子孢子挑战。多肽/MHC相互作用的相关性 在体外具有功能免疫原性的体内将验证使用 开发可诱导的疟疾疫苗的途径 人的体液免疫和细胞免疫的保护水平 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.