负载自体肿瘤抗原的成熟DC体外联合小剂量IL-2及IL-12增强CTL抗胃癌活性研究  

DENDRITIC CELLS LOADED WITH AUTOLOGOUS TUMOR LYSATES AUGMENT CYTOLYTIC ACTIVITY OF CTL AGAINST GASTRIC CANCER IN COMBINATION WITH LOW DOSE IL -2 AND IL -12 EX VIVO

作　　者：谢绍建[1] 崔乃鹏[1] 张俊岭[1] 陈小贺[2] 张峰[2] 蔡建辉[1] 

机构地区：[1]河北医科大学第二医院生物治疗肿瘤科,河北石家庄050000 [2]河北医科大学第二医院外科,河北石家庄050000

出　　处：《河北医科大学学报》2011年第6期625-629,F0003,共6页Journal of Hebei Medical University

基　　金：河北省科技厅博士基金项目(04547002D-9);河北省卫生厅重大攻关项目(2003001)

摘　　要：目的探讨在体外条件下负载自体胃癌细胞裂解物的成熟DC(matured dendritic cells loaded withautologous tumor lysates,TL-mDC)联合小剂量白细胞介素-2(interleukin-2,IL-2)及白细胞介素-12(interleukin-12,IL-12)增强细胞毒性T淋巴细胞(cytotoxic T lymphocytes,CTL)抗胃癌免疫效应。方法建立短期培养的原代胃癌细胞系作为靶细胞。负载自体胃癌细胞裂解物的成熟DC联合应用小剂量IL-2及IL-12体外刺激自体T细胞,制备胃癌特异性CTL。应用淋巴细胞增殖实验检测联合刺激后T细胞的增殖能力;应用细胞计数试剂盒(cell counting kit-8,CCK-8)试剂盒检测CTL对肿瘤的特异性杀伤活性;应用逆转录聚合酶链反应法测定CTL.培养液中干扰素γ(interferon-γ,INF-γ的分泌水平。结果 TL-mDC体外联合小剂量IL-2、IL-12刺激组(组1),与单纯TL-mDC刺激组(组2)比较,具有高效刺激自体T细胞增殖的能力(P<0.01);2组分别与T细胞混合培养1周后,所获CTL上清液中INF-γ的浓度分别为(3200.00±172.57)ng/L(组1)、(2700.00±83.18)ng/L(组2),差异有统计学意义(P<0.01);CTL(组1)对自体胃癌细胞的杀伤率显著高于CTL(组2)(P<0.05),且CTL,(组1及组2)对自体胃癌细胞的杀伤率显著高于对2株异体胃癌细胞的杀伤率(P<0.01)。结论 TL-mDC体外联合小剂量IL-2及IL-12有效提高了CTL对自体胃癌细胞的特异杀伤活性。Objective The efficiency of adoptive immunotherapy against gastric cancer induced by matured dendritic cells loaded with autologous tumor lysates （TL - mDC） was investigated in combination with low dose IL - 2 and IL - 12 ex vivo. Methods Primary gastric cancer cells, as targets in the present study, were prepared by short - term culturing. TL - mDC was subjected to autologous gastric lysate to generate activated T lymphocytes in combination with low dose IL -2 and IL - 12 ex vivo. Proliferation abilities of co - stimulated naive T lymphocytes were tested by mixed leukocyte response （MLR） assay. The specific cytotoxicity of eytotoxic T lymphoeytes （CTL） for tumor was tested by cellcounting kit - 8 （ CCK - 8 ）. The concentrations of interferon - γ （ INF - γ） in supernatant of activated T lymphocytes were examined by using ELISA. Results A remarkable increased proliferation ability of autologous T lymphoeytes was observed in the first group co - cultured with TL - mDC plus low dose IL - 2 and IL- 12 than that in the second group co-cultured with TL-mDC alone （P〈0.01）. The concentration of INF - γ/in the supernatant of activated T lymphocytes in the first group was significantly higher, as compared with that in the second group [ （3200.00 ± 172.57 ） ng/L us （2700.00 ± 83.18 ） ng/ L,P 〈0.01 ]. More powerful tumor specific cytotoxicity against autologous tumor cells was exhibited by the activated T lymphocytes in the first group than those in the second group （ P 〈 0.05 ）, and both two group CTLs had strikingly higher specific cytotoxicity against autologous tumor cells than against allogeneic tumor cells （ P 〈 0.01 ; P 〈 0. 01 ）. Conclusion TL - mDC in combination with low dose IL - 2 and IL - 12 might mediate CTL to generate significantly enhanced tumor speeific cytolytic activity against autologous gastric eancer cells.

关 键 词：胃肿瘤 树突细胞 T淋巴细胞细胞毒性. 

分 类 号：R735.2[医药卫生—肿瘤]