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目的:观察转染4-1BBL重组腺病毒载体的小鼠前列腺癌细胞瘤苗体外诱导抗肿瘤免疫作用及机制。方法:利用复制缺陷型腺病毒AdEasy-1系统,通过基因重组技术构建Ad-m4-1BBL及Ad-eGFP。将Ad-m4-1BBL和Ad-eGFP感染小鼠前列腺癌细胞RM-1,以丝裂霉素C(MMC)处理,制成肿瘤细胞疫苗(TCV,TCV-Ad-eG-FP,TCV-Ad-m4-1BBL),经体外与同系小鼠脾淋巴细胞共同培养,ELISA法检测脾细胞分泌细胞因子(IL-2,INF-γ)的影响,CCK-8试剂检测脾淋巴细胞特异性杀伤活性。结果:转染4-1BBL基因的小鼠前列腺癌细胞瘤苗TCV-Ad-m4-1BBL高表达4-1BBL蛋白,培养上清中细胞因子IL-2[(180.24±2.22)pg/ml]和INF-γ[(1 512.46±23.64)pg/ml]表达水平均明显高于TCV和TCV-Ad-eGFP组(P<0.05),TCV-Ad-m4-1BBL瘤苗诱导RM-1细胞特异性杀伤率[(34.24±2.64)%],相比TCV-Ad-eGFP[(14.65±3.21)%]和TCV[(9.82±1.48)%],差异有显著性(P<0.05)。但针对Hepal-6细胞,3种瘤苗均无效。结论:表达4-1BBL的小鼠前列腺癌细胞瘤苗能诱导有效的抗肿瘤免疫反应。
OBJECTIVE: To investigate the anti-tumor immunity induced by 4-1BBL recombinant adenovirus vector and its mechanism in vitro. Methods: Ad-m4-1BBL and Ad-eGFP were constructed by recombinant adenovirus AdEasy-1 system. TCV-Ad-eG-FP and TCV-Ad were transfected into mouse prostate cancer cells RM-1 infected with Ad-m4-1BBL and Ad-eGFP by mitomycin C (MMC) -m4-1BBL). The spleen lymphocytes were co-cultured in vitro and in vivo. The secretion of cytokines (IL-2, INF-γ) by spleen cells was detected by ELISA. The specific cytotoxic activity of spleen lymphocytes was detected by CCK-8 . Results: 4-1BBL protein was highly expressed in mouse prostate cancer cell vaccine TCV-Ad-m4-1BBL transfected with 4-1BBL gene. The cytokine IL-2 in the culture supernatant was (180.24 ± 2.22) pg / ml] The expression of INF-γ [(1 512.46 ± 23.64) pg / ml] was significantly higher than TCV and TCV-Ad-eGFP groups (P <0.05). The TCV-Ad-m4-1BBL vaccine induced RM- The killing rate [(34.24 ± 2.64)%] was significantly higher than that of TCV-Ad-eGFP [(14.65 ± 3.21)%] and TCV [(9.82 ± 1.48)%] (P <0.05). However, for Hepal-6 cells, all three tumor vaccines were ineffective. CONCLUSIONS: Mouse prostate cancer cell vaccine expressing 4-1BBL induces potent anti-tumor immune responses.