基于热休克蛋白的MUC1治疗性疫苗的研究

发布时间：2018-02-02 05:54

本文关键词： 结核分支杆菌热休克蛋白65 MUC1 治疗性疫苗　出处：《中国人民解放军军事医学科学院》2005年硕士论文　论文类型：学位论文

【摘要】：MUC1是Mucins粘蛋白中的一种,存在于正常腺管上皮及其来源的肿瘤细胞表面。它是一种大分子的糖蛋白,由多肽核芯和侧枝糖链构成,其多肽核芯的胞外段含有20个氨基酸的串联重复序列(Variable numbers tandem repeats,VNTRs)。在多种肿瘤中,MUC1异常表达,主要表现为表达量增高,细胞表面极性分布的改变和结构改变等,成为免疫细胞攻击的靶点。研究发现MUC1核芯肽的PDTRP表位不仅能被MUC1抗体识别,而且也能被CTL细胞识别和杀伤,且不受MHC的限制。由于MUC1具有上述特点,成为理想的抗肿瘤靶分子。本课题的目的即是开发一种基于热休克蛋白的针对非小细胞肺癌(Non small cell lung cance,NSCLC)的MUC1治疗性疫苗。近年来发现,热休克蛋白—多肽复合物可通过受体(CD91)介导的内吞作用被巨噬细胞吞噬,然后经过蛋白酶体降解,最终进入内源性抗原递呈途径,从而激活细胞免疫。我们选择结核分支杆菌热休克蛋白65(HSP65)与鼠BP24或人BP25基因融合来作为肿瘤的治疗性疫苗。它可以把BP24或BP25蛋白呈递给MHC-I抗原呈递途径,进而激活细胞免疫。我们通过克隆得到鼠BP24和人BP25基因,构建了HSP65-BP24-Ztag和HSP65-BP25-Ztag的pET28a表达载体,并在大肠杆菌中得到了高效表达。目的蛋白经硫酸铵沉淀、阳离子层析、疏水层析获得了纯度超过90%的样品。经纯化的蛋白作为非小细胞肺癌的治疗性疫苗。应用HSP65-BP24-Ztag融合蛋白在小鼠Lewis肺癌细胞模型上进行免疫治疗和预防试验。结果证明,预防免疫可以使小鼠成瘤小于对照组;治疗免疫可抑制小鼠肿瘤生长,生存期延长。
[Abstract]:MUC1 is one of Mucins mucin , which is present in the surface of tumor cells of normal glandular epithelium and its origin . It is a large molecule glycoprotein composed of polypeptide core and side branch sugar chain . The extracellular portion of the polypeptide core contains 20 amino acids in tandem repeats ( Variable numbers tandem repeats , VNTR ) . MUC1 expression in various tumors is mainly expressed as an increase in the expression level , changes in the polarity distribution of the surface of the cell and structural changes , etc . , and becomes a target for immune cell attacks . In recent years , heat shock protein - polypeptide complexes can be phagocytosed by macrophages by endocytosis mediated by receptor ( CD91 ) and then degraded by proteasome to finally enter endogenous antigen presenting pathway , thus activating cell immunity . We chose Mycobacterium tuberculosis heat shock protein 65 ( HSP65 ) to fuse with murine BP24 or human BP25 gene as a therapeutic vaccine for tumors . The expression vector pET28a of HSP65 - BP24 - Ztag and HSP65 - BP25 - Ztag was constructed by cloning mouse BP24 and human BP25 gene . The purified protein was used as a therapeutic vaccine for non - small cell lung cancer . The purified protein was used as a therapeutic vaccine for non - small cell lung cancer .

【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392.1

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