| 摘要: |
| 目的观察人乳头瘤病毒（HPV）-16E7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-16E7载体转染DC后其细胞表型，流式细胞术检测HPV-16E7负载DC与CIK细胞共培养后CIK细胞的细胞表型。ELISA法检测HPV-16E7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-16E7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-16E7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-16E7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。 |
| 关键词: 树突状细胞 细胞因子诱导的杀伤细胞 人乳头瘤病毒 宫颈癌 肿瘤杀伤效应 |
| DOI：10.12056/j.issn.1006-2785.2018.40.24.2018-1915 |
| 分类号: |
| 基金项目:浙江省科技计划项目（2016C33241） |
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| HPV-16 E7-pulsed dendritic cells promote cytotoxicity of CIK cells against cervical carcinoma cells |
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| Abstract: |
| 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. |
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