过继肿瘤特异性淋巴细胞联合mIL-21瘤苗抗肿瘤效应研究

目的 在环磷酰胺(Cy)所致小鼠淋巴细胞急剧减少状态下,过继mIL-21转染的瘤苗细胞(mIL-21-Sp2/0)免疫致敏的同系小鼠淋巴细胞,联合mIL-21-Sp2/0瘤苗免疫,探讨过继免疫抗瘤效应机制.方法 用Cy 100 mg/kg预处理BALB/c小鼠2 d后,以灭活mIL-21瘤苗免疫的同系小鼠脾及淋巴结的淋巴细胞作为肿瘤抗原特异性淋巴细胞过继给Cy预处理鼠,同时用mIL-21瘤苗免疫,7 d后以野生型Sp2/0细胞攻击,观察小鼠的肿瘤生长情况.流式细胞仪(FCM)分析CD4+、CD8+、CD4+CD25+T细胞等亚群的变化,分别以CFSE和7-AAD标记,FCM检测淋巴细胞增殖能力和效应细胞的细胞毒活性,用ELISPOT检测分泌IFN-γ的淋巴细胞数量.结果 与对照组相比,Cy预处理小鼠接受过继效应细胞及mIL-21瘤苗免疫后,可有效地抵抗野生型Sp2/0细胞的攻击.过继的肿瘤特异性淋巴细胞的体内增殖能力和体外杀伤活性显著增强,分泌IFN-γ效应细胞的数量显著增高.结论 在小鼠淋巴细胞减少期,过继mIL-21瘤苗免疫致敏的肿瘤抗原特异性淋巴细胞并同时给予mIL-21瘤苗免疫,利于输入的效应细胞及自身免疫细胞的增殖和特异性抗肿瘤功能的形成与维持.

Anti-tumor mechanisms of lymphopenic mice transferred with tumor-specific lymphocytes and immunized with mIL-21 tumor vaccine

Objective To explore the mechanism of anti-tumor effects of transferring tumor-specif-ic lymphocytes obtained from pre-immunized BALB/c mice with inactive rolL-21 tumor vaccine (mIL-21-Sp2/0)to syngeneic mice, associated with mIL-21 tumor vaccine immunization, in the condition of cyclo-phosphamide (Cy)-induced lymphopenia. Methods Activated lymphocytes of spleen and lymph nodes ob-tained from pre-immunlzed syngeneic mice with irradiated mIL-21-Sp2/0 cells were infused into BALB/cmice treated with Cy 2 days before, subsequently vaccinated with mlL-21 tumor vaccine, after 7 days, chal-lenged with Sp2/0 tumor cells, observed the growth of tumor of mice. T lymphocyte subsets differentiation was measured by flow cytometry (FCM) analysis. The proliferation and cytotoxie activities of activated lym-phocytes were analyzed by FCM, respectively, staining with CFSE and 7-AAD. The number of IFN-γ-secre-ting cells was evaluated by ELISPOT. Results The lymphopenic mice were transferred with activated lym-phocytes and inoculated with raiL-21 tumor vaccine might provide superior anti-tumor immunoprotection, re-tard tumor growth of the mice. The proliferating capabilities and killing rate of transferred tumor Ag-specific lymphocytes enhanced obviously, the number of IFN-γ-secreting cells was significantly higher compared with the control groups. Conclusion Under Cy-induced lymphopenia condition, tumor Ag-specific lymphocytes sensitized by raiL-21 tumor vaccine were transferred to mice and immunized with mlL-21 tumor vaccine at the same time, benefit the proliferation of transferred effective cells and immune cells itself, assist to form and sustain special anti-tumor effects.