重组蛋白PSCA-HSP70的免疫活性及抗肿瘤效应观察  

Evaluation of immune effect of recombinant fusion protein targeting the prostate stem cell antigen based on PSCA and HSP70

作　　者：董磊[1] 刘娟[2] 马红雨[1] 全首祯[1] 吕建晓[1] 李莹[1] 朱美财[1] 

机构地区：[1]空军总医院临床检验中心,北京100142 [2]空军总医院输血科,北京100142

出　　处：《解放军医学杂志》2014年第9期714-719,共6页Medical Journal of Chinese People's Liberation Army

基　　金：空军总医院基金(kz2013021)~~

摘　　要：目的探讨重组前列腺干细胞抗原(PSCA)-热休克蛋白70(HSP70)融合蛋白对前列腺肿瘤模型小鼠的免疫激活作用及抗肿瘤活性。方法健康雄性C57BL/6小鼠25只,随机均分为PSCA组、HSP组、PSCA+HSP组、PSCAHSP组及PBS组(n=5)。小鼠皮下注射1×106个前列腺肿瘤细胞(RM-PSCA/Luc细胞),3d后分别采用相应重组蛋白进行免疫(100μg/只),2周后采用相同剂量加强免疫1次,观察肿瘤生长及小鼠存活状况。加强免疫后10d处死动物,采用ELISPOT法和流式细胞术检测PSCA特异性T细胞反应,ELISA法检测PSCA特异性抗体水平。结果 ELISPOT法检测显示,PSCA-HSP组分泌IFN-γ的CD8+T细胞数明显高于其他各组(P<0.05);流式细胞术检测结果显示,PSCA-HSP组CD8+/IFN-γ+T细胞数明显高于其他各组(P<0.05);ELISA检测显示,PSCA、PSCA+HSP、PSCA-HSP均可诱导产生PSCA特异的体液免疫反应,且3组间抗体水平差异无统计学意义(P>0.05)。PSCA-HSP组肿瘤体积明显小于其余各组,小鼠存活时间明显长于其余各组(P<0.05)。结论 HSP70是一种效应显著的分子伴侣,重组PSCA-HSP70融合蛋白在前列腺癌的治疗中具有重要的潜在应用价值。Objective To explore the immune effect and antitumor activity of recombinant prostate stem cell protein （PSCA） and heat shock protein 70 （HSPT0） in a murine model of prostate cancer. Methods Twenty-five healthy male C57BL/6 mice were randomly divided into 5 groups （5 each）： PSCA, HSP, PSCA＋HSP, PSCA-HSP and control group. Mice in the first 4 groups were vaccinated with the corresponding proteins, and those in control group were faked with injection of phosphate buffer saline （PBS）. After immunization with recombinant proteins, the PSCA-specific cellular immune responses were monitored with ELISPOT, intracellular cytokine staining assay, and flow cytometry, and ELISA assay was used to detect humoral immune responses. The tumor growth and survival ofvaccined mice were observed. Results ELISPOT revealed that the mice in PSCA-HSP group generated much more IFN-γ spot-forming cells than those in other groups （P〈0.0S）, and they could generate strong anti-PSCA antibody response. Results of flow cytometry showed that the number of CD8^＋/IFN-γ^＋ T cells was significantly higher in PSCA- HSP group than that in other groups （P〈0.05）. ELISA results revealed that all the mice in PSCA, PSCA＋HSP and PSCA-HSP group were induced to generate the PSCA-specific humoral immune response, and no statistical difference was found on the antibody levels among the three groups. Animal experiment showed that PSCA-HSP could inhibit the growth of PSCA-expressing tumors and prolong the survival time of vaccinated mice. Conclusion HSP70 is a chaperone with significant effect for protein vaccines, and the recombinant fusion protein PSCA-HSP70 could be of potential value for prostate cancer treatment.

关 键 词：前列腺干细胞抗原 HSP70热休克蛋白质类 肿瘤 实验性 

分 类 号：R737.14[医药卫生—肿瘤]