截短性HSP110抗原肽复合物免疫小鼠的CTL活性研究  

Activity of cytotoxic lymphocyte in BALB/c mice immunized by truncated HSP110-ECD

作　　者：徐煌[1] 韩冬[1] 

机构地区：[1]嘉兴学院医学院生物化学教研室,浙江嘉兴314001

出　　处：《中华肿瘤防治杂志》2015年第17期1368-1372,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：浙江省科技厅公益技术应用研究项目(2015C33106);嘉兴市科技局一般科研项目(2014AY21038)

摘　　要：目的观察基因工程来源的截短性热休克蛋白110(truncated heat shock protein110,tHSP110)抗原肽复合物免疫BALB/c小鼠在体外诱导细胞毒性T淋巴细胞(cytotoxic lymphocyte,CTL)的活性。方法基因工程来源的tHSP110在体外与人类表皮生长因子受体2(human epidermal growth factor receptor 2,HER2/neu)的胞外区(extracellular domain,ECD),非共价耦联方法构建tHSP110-ECD抗原肽复合物,并通过免疫共沉淀方法鉴定。分3次(每周1次)用HSP110、tHSP110、HSP110-P106-114(全长HSP110与HER2/neu ECD的一个CTL表位肽组成的复合物)、tHSP110-P106-114和HSP110-ECD复合物免疫BALB/c小鼠,末次免疫1周后,采用酶联免疫斑点(enzyme-linked immunospot,Elispot)法对T细胞分泌干扰素-γ(interferon-γ,IFN-γ)的水平进行检测,通过颗粒酶释放法检测tHSP110-ECD诱导的杀伤性T细胞的抗肿瘤效应。结果 T细胞IFN-γ分泌测定结果显示,全长HSP110组蓝色斑点数量为(17.50±3.89)个,tHSP110组为(17.75±3.54)个,HSP110-P106-114组为(72.16±4.22)个,tHSP110-P106-114组为(70.16±2.29)个,HSP110-ECD组为(90.38±5.85)个,tHSP110-ECD组为(88.38±6.14)个。tHSP110-P106-114组与tHSP110-ECD组分泌IFN-γ的小鼠脾细胞数目差异有统计学意义,P<0.05;tHSP110-ECD组与HSP110-ECD组相比,差异无统计学意义,P=0.741。特异性CTL检测的结果显示,HSP110组的杀伤率为(9.63±1.67)%,tHSP110组为(9.00±1.60)%,HSP110-P106-114组为(36.38±2.62)%,tHSP110-P106-114组为(38.50±4.37)%,HSP110-ECD组为(72.88±3.68)%,tHSP110-ECD组为(73.38±3.66)%。tHSP110-P106-114组与tHSP110-ECD组靶细胞杀伤率差异有统计学意义,P<0.05;tHSP110-ECD组与HSP110-ECD组相比差异无统计学意义,P>0.05。结论 tHSP110-ECD复合物作为肿瘤疫苗可刺激T淋巴细胞增殖为CTL,诱发机体的细胞毒性T细胞免疫反应,tHSP110具有同全长HSP110相同的结合大分子抗原的能力和激活CTL反应的能力。OBJECTIVE To observe the specific cytotoxic T lymphocyte（CTL） induced by truncated heat shock protein 110-extracellular domain of human epidermal growth factor receptor 2 （tHSP110-ECD） complexes immunization in BALB/c mice. METHODS Female BALB/c mice were immunized with HSP110, HSP110-P106-114 （full length HSP110 and one CTL peptide of HER2/neu ECD complex）,tHSP110-P106-114 ,tHSP110-ECD, HSP110-ECD complexes 3 times at in- tervals of 3 weeks. One week after the last immunization, the frequency of Interferon-y（IFN-γ） producing cells and the CTL activity was elevated. RESULTS Treatment with HSP110,tHSP110,HSP110-P106-114 ,tHSP110-P106-114 ,HSP110 ECD, tHSP110-ECD complexes respectively, the number of spot-forming cells of the groups were （17.50 ± 3.89）, （17. 75 ± 3.54）, （72.16±4.22）, （70.16±2.29）, （90.38±5.85） and （88.38±6.14）. The number of spot-forming cells in the tH- SP110-ECD group was higher than the tHSP110-P106-114 （P〈0.05）. The number of IFN-γ secreting spleen lymphocytes in the HSP110 ,tHSP110, HSP110-P106-114, tHSP110-P106-114, HSP110-ECD, tHSP110-ECD complexes group were （9. 63 ± 1.67）%,（9.00±1.60）%,（36.38±2.62）%,（38.50±4.37）%, （72. 88±3. 68）% and （73.38±3.66）% respectively. The number of IFN-γ secreting spleen lymphocytes in the tHSP110-ECD group was significantly higher than that in thetHSP110-P106-114 group（P〈0.05）. There was no difference between tHSP110-ECD group and HSP110-ECD group in specific CTLs activity （P=0. 910） and the Elispot analysis （P= 0. 741）. CONCLUSIONS These findings support that this truncated HSP110-ECD vaccine has the same ability of binding tumor antigen and can induce specific CTLs activity. It plays an important role in host anti-tumor immunity by stimulating T lymphocyte proliferation,and inducing CTL production, tHSP110 has the same chaperone activity with HSP110.

关 键 词：截短性HSP110 T淋巴细胞 免疫应答 疫苗 

分 类 号：R73-36[医药卫生—肿瘤]