人41BBL抗CD20融合蛋白增强抗CD3抗CD20双功能抗体的细胞毒作用及其机制研讨.pdfVIP

所 院 ： 血液学研究所 姓 名 ： 刘荣 指导教师： 熊冬生教授 导师小组： 熊冬生朱惠芳杨纯正许元富 学科专业： 肿瘤药理学 研究方向： 基因工程抗体 完成日期： 二O—o年五月 二零一零年五月 一令一奄，牛直月 目 录 ··········································-·············3 ························································8 4．1BBL胞外区／抗CD20融合蛋白表达载体的构建···12 ······································-················12 ·····················································-·13 ·······················································23 ·······················································27 细胞毒作用及其机制研究············…················28 l实验材料·························································28 2实验方法························································29 3实验结果··························································34 讨 论···························································42 结 论···························································56 参考文献··········……········…······…················……·57 论文综述···························································66 附录一、英文缩略词表····················…·····…·········……曝9 附录二、溶液配制·……··…····…··········…·············……91 致 谢···························································96 信号即第二信号。T细胞激活诱导阶段若缺乏共刺激信号，会引起T细胞通过活化诱 导的细胞死亡途径凋亡和克隆特异性无反应性，进而导致抗肿瘤免疫功能低下，而 通过低表达共刺激分子降低免疫原性，使得识别它的抗原提呈细胞或淋巴细胞得不 到充分的活化信号，是肿瘤细胞免疫逃逸的重要机制之一，临床实践中，这类肿瘤 的治疗效果和预后不佳。：共刺激信号途径在T细胞的活化和增殖过程中起着重要作 用。CD28／B7分子是研究最多的协同刺激分子，认为是初始T细胞激活发生最主要的 达在T细胞和抗原提呈细胞上，被确定有助于扩增和多元化T细胞反应，深入增强和 延伸T细胞激活的功能，许多动物实验表明干预4—1BB／4—1BBL共刺激途径可调节T细 胞和抗原呈递细胞的功能产生抗肿瘤免疫作用，为肿瘤的免疫治疗提供了新的靶 点。 CD20主要表达在前B细胞和成熟B细胞及95％以上的NHL淋巴瘤细胞，是可以用来 作为肿瘤定向免疫治疗的抗原。为了特异性定向肿瘤细胞和激活肿瘤特异性T细胞， 抗肿瘤免疫性能够被诱导和恢复，利用双特异抗体针对肿瘤相关抗原与效应细胞相 连，共同作用于肿瘤细胞，实现肿瘤的靶向治疗，我们已成功构建了抗CD3／