HPV-16 E7负载DC激活CIK细胞增强宫颈癌细胞杀伤效应的研究

HPV-16 E7-pulsed dendritic cells promote cytotoxicity of CIK cells against cervical carcinoma cells

目的观察人乳头瘤病毒(HPV)-16 E7特异性高表达的树突状细胞(DC)对细胞因子诱导的杀伤(CIK)细胞激活及宫颈癌细胞杀伤效应的影响。方法采用流式细胞术检测HPV-16 E7载体转染DC后其细胞表型,流式细胞术检测HPV-16 E7负载DC与CIK细胞共培养后CIK细胞的细胞表型。ELISA法检测HPV-16 E7负载DC-CIK细胞、阴性对照负载DC-CIK细胞、DC-CIK细胞、单纯CIK细胞中CIK细胞分泌的IL-12、IFN-γ、TNF-α等细胞因子的表达水平,乳酸脱氢酶(LDH)释放法检测CIK细胞对宫颈癌Hela细胞的杀伤活性。结果 HPV-16 E7负载可促使DC成熟,细胞表面标志物CD83、CD86及HLA-DR的表达上调,CD14表达下调。经过HPV-16 E7负载DC共培养的CIK细胞,CD3^+CD8^+、CD3^+CD56^+细胞比例均上调,细胞因子IL-12、IFN-γ、TNF-α的水平亦均上调,CIK细胞表现出更强的宫颈癌Hela细胞杀伤活性。结论 HPV-16 E7负载DC不仅能激活CIK细胞,还可增强CIK细胞的宫颈癌细胞杀伤效应。这为宫颈癌的细胞免疫治疗提供新思路。

Objective To investigate the effect of human papillomavirus(HPV)-16 E7-pulsed dendritic cells(DC) on cytotoxicity of cytokine-induced killing(CIK) cells against cervical cancer cells. Methods The phenotype of DCs transfected with pcDNA3. 1-HPV-16 E7 was detected by flow cytometry( FCM). CIK cells were co-cultured with HPV-16 E7-pulsed DCs, the surface markers on CIK cells were detected by FCM, and the levels of cytokines IL-12, IFN-γ, TNF-α secreted by CIK cells were measured by ELISA. The cytotoxicity CIK cells, DC-CIK cells and HPV-16 E7-pulsed DC-CIK cells against to cervical cancer Hela cells was detected by LDH release method. Results Over-expression of HPV-16 E7 promoted DCs maturation, increased the levels of CD83, CD86 and HLA-DR, decrease CD14 expression. Co-culture of the CIK cells with HPV-16 E7-pulsed DCs significantly increased the positive rate of CD3+CD56+ cells, and the levels of IL-12, IFN-γ and TNF-α; and had the highest cytotoxic effect on Hela cells. Conclusion DCs pulsed by HPV-16 E7 can activate CIK cells and enhance its cytotoxicity to cervical carcinoma cells, which provides an experimental basis for cell immunotherapy of cervical carcinoma.