《t细胞表位的研究总结》ppt模板课件.pptVIP

Add the author and the accompanying title;第三章 T细胞表位 Antigen ;此文档后面有赠送常用PPT图标，方便大家修订排版编辑;第一部分T细胞表位的基本概念;抗原结合价 Antigenic valence 能与抗体分子结合的抗原表位的总数。;2、抗原表位的类型;T细胞表位与B细胞表位;3、共同抗原表位(common epitope);; 抗原肽和MHC相互作用的分子基础 * 锚定残基（anchor residue） 在抗原肽-MHC分子复合物中，抗原肽的两个或两个以上专司和MHC分子结合的氨基酸残基称为锚定残基。 ;;MHC I 类分子胞外区的结构;抗原肽和HLA相互作用的分子基础;T细胞表位的研究方法 1.多肽片段的来源 降解；表达；人工合成 2. MHC分子的结合 直接结合；预测 3. 抗原特异性T细胞的来源 感染或者免疫动物或病人 筛选方法 细胞系的建立 4. 检测指标 活化；增殖；细胞因子；杀伤作用 ;T细胞表位的研究意义 1.检测 刺激细胞使用 Tetramer技术 MHC 2. 疫苗 表位疫苗 3. 治疗 ; 第二部分 SARS-CoV S蛋白T细胞表位的研究 ;1. SARS-CoV S DNA 疫苗;2.免疫方法;3. 单细胞悬液的准备;4. SARS-CoV S蛋白多肽;;Fig. 2. Verification of potential SARS CoV S epitopes Potential SARS CoV S epitopes P50, P51, P59 and P60 in pool 3, and P141, P144, P151 and P152 in pool 8 were used to stimulate splenocytes from SARS CoV S DNA immunized BALB/c mice. IFN-γ production was detected by ELISA (A) and ELISPOT (B), respectively. Each symbol represents the results of an individual experiment (n=3~7). In addition, intracellular cytokine staining (C) was performed to determine CD4+ or CD8+ T cell population. “0” represents the non-peptide stimulated control. Numbers at the corner in each sample represent the percentage of positive cells. Representative results of three independent experiments were shown. ;Fig. 3. Identification of new synthetic 10aa CD4 and CD8 epitopes The overlapped amino acids between P50 and P51 (N50), and between P59 and P60 (N60) were synthesized. Peptides N50 and N60 were used to stimulate splenocytes from SARS CoV S DNA vaccine-immunized mice. P50 and P60 were used as positive controls, respectively. ELISA (A) and ELISPOT (B) were performed as described above. Furthermore, N50 and 60 were serial diluted to stimulate splenocytes from the DNA immunized mice, ELISPOT (C) was performed to detect numbers of antigen specific IFN-γ producing cells. Experiments were carried out in duplicate and representative results were shown. “0” represents non-peptide-cultured neg