【摘要】 目的研究gp96多肽复合物树突状细胞(DC)疫苗特异性抗胃癌的免疫反应。方法应用层析技术从胃癌组织中提取gp96多肽复合物,制备gp96多肽复合物DC疫苗;采用流式细胞仪检测gp96多肽复合物DC疫苗表面分子的表达;ELISA检测gp96多肽复合物DC疫苗致敏的效应 T淋巴细胞分泌IFN-γ和IL-10的水平;51Cr释放实验检测gp96多肽复合物DC疫苗诱导的特异性对胃癌细胞的杀伤效应。结果gp96多肽复合物DC疫苗表面分子CD1α(79．3±4．1)％、CD80 (84．3±2．4)％、CD83(85．7±3．2)％和HLA-DR(83．4±2．9)％的表达显著增高;对原代培养胃癌细胞的杀伤率(58．5±10．7)％显著高于对SGC 7901细胞的杀伤率(24．0±4．2)％,两者相比差异有统计学意义,P<0．01;对HepG2细胞(13．8±2．8)％则无明显杀伤作用。DC疫苗诱导的效应T淋巴细胞分泌IFN-γ(2875±178)pg／ml的量明显增加,分泌IL-10(36±7)pg／ml的量显著降低。结论 gp96多肽复合物DC疫苗能诱导出较强的对自体胃癌细胞的杀伤作用,且具有高度特异性。更多还原

【Abstract】 Objective To investigate the anti-gastric cancer effects of dendritic cells tumor vaccine loaded with heat shock protein gp96 polypeptide complex. Methods Con A sepharose affinity chromatography and mono Q ion exchange chromatography were applied to purify gp96 polypeptide complex from gastric cancer tissues and DC was loaded with gp96 polypeptide complex to prepare tumor DC vaccine. Fluorescence-activated cell sorter (FACS) was used to detect surface molecules expression of dendritic cells. Enzyme-linked immunosorbent assay ( ELISA) was used to detect IL-10 and interferon-γ (IFN-γ) of the supernatant secreted by activated T lymphocytes and 51Cr release test was performed to evaluate the gp96 peptide specific CTL response. Results DCs derived from patient’s peripheral blood mononuclear cells loaded with gp96 polypeptide complex had high express surface molecule of GD1α (79. 3 ±4. 1)% , CD80 (84. 3 ± 2. 4) % , CD83 (85. 7 ± 3. 2) % and HLA-DR( 83.4 ± 2. 9) %. The effector T lymphocytes induced by DCs vaccine loaded with gp96 polypeptide complex had rised killing effect to primary culture gastric tumourcells(58.47±10. 71)% when compared with SGC 7901 cells(23. 98 ±4. 16)%. The level of IFN-γ (2875 ± 177.66 pg/ml) significantly increased and that of IL-10 (36 ± 6.72 pg/ml) decreased. Conclusion Autologous heat shock protein gp96 polypeptide complex/DC vaccine induces strong peptide complex specific CTL response with high specificity.更多还原