C6细胞热休克蛋白抗原肽复合物提纯及抑瘤作用研究

目的提纯大鼠脑胶质瘤C6细胞热休克蛋白抗原肽复合物(HAC),免疫SD大鼠,观察HAC的抑瘤作用。方法采用免疫亲和层析方法提纯大鼠脑胶质瘤C6细胞HAC,免疫20只大鼠为实验组,以另20只大鼠作为对照组,于免疫后1 w,采用立体定向脑内接种方法,以C6细胞攻击两组大鼠,于肿瘤细胞攻击后第二周,取两组动物外周静脉血,测定外周静脉血淋巴细胞计数,并应用流式细胞仪技术测定外周血中CD3 /CD4 和CD3 /CD8 T淋巴细胞的比例。观察饲养过程中实验动物出现的症状、体征和第四周实验动物存活率。于第四周处死存活动物,取脑组织进行HE染色病理组织学检查,并用免疫组化方法分析脑胶质瘤浸润区T淋巴细胞分布情况。结果实验组大鼠外周血淋巴细胞计数显著高于对照组(P<0.01),CD3 /CD4 和CD3 /CD8 T淋巴细胞的比例实验组均显著高于对照组(P<0.01)。实验组动物症状出现时间显著晚于对照组动物(P<0.01),实验组动物四周末存活率显著高于对照组(P<0.05)。实验组胶质瘤局部浸润的CD3 和CD4 细胞数均显著高于对照组(P<0.01),实验组胶质瘤局部浸润的CD8 细胞数与对照组比较无显著差异(P>0.05),实验组胶质瘤局部浸润T淋巴细胞CD4 /CD8 显著高于对照组(P<0.01)。结论C6细胞中HAC可以诱导大鼠产生对C6细胞的细胞免疫,提高大鼠存活率。

Research on tumor-suppressive effect of HAC purified from C6 glioma cells

Objective To study the tumor-suppressive effect of purified heat shock protein-antigen complex(HAC) in SD rats immunized with HAC.Methods Immuno-affinity chromatography was used to purify HAC from C6 cells.Twenty rats were immunized with HAC to form the experimental group,and other 20 non-immunized rats served as the control group.One week after immunization,C6 cells were inoculated into the brains of the rats by(stereotaxic) method.Two weeks later,peripheral venous blood was drawn,total lymphocyte count was done,and the rates of CD3~( )/CD4~( ), CD3~( )/CD8~( )T lymphocytes were measured by flow cytometry.Four weeks after inoculation,the symptoms manifested by the animals were observed and their survival rates were estimated.All the rats were(killed) and their brain tissues were sent for histopathologic and immunohistochemical examinations.Results The(number) of total peripheral blood lymphocytes and CD3~( )/CD4~( ),CD3~( )/CD8~( ) T lymphocytes of the experimental group rats were significantly higher than those in the control group rats(P<0.01).The quantity of CD3~( ) and CD4~( )T(lymphocytes) that infiltrated into the tumor tissues of the experimental group rats was significantly larger than that of the control group rats(P<0.01).There was no significant difference in CD8~( ) T lymphocyte infiltration between these two groups.The ratio of CD4~( )/CD8~( ) T lymphocytes of the experimental group rats was significantly higher than that of the control group rats(P<0.01).The survival rate of the experimental group rats was significantly higher than that of the control group rats(P<0.05).Conclusion Rats immunized with HAC purified from C6 cells(develop) immuno-resistance against C6 glioma cells,and their survival rates are improved.