经卡介苗活化的负载PANC1裂解产物的树突状细胞疫苗体外抗胰腺癌作用  

In vitro anti-tumor effect of autologous mixed lymphocyte primed by BCG activated dendritic cells based PANC1 lysate

作　　者：杨德红[1] 刘文佳[1] 陈敏[1] 吴育美[1] 邹晓平[1] 

机构地区：[1]南京大学医学院附属鼓楼医院消化科,南京210008

出　　处：《中华胰腺病杂志》2010年第4期267-271,共5页Chinese Journal of Pancreatology

摘　　要：目的探讨负载胰腺癌细胞株PANCl裂解产物（tumor lysate，TL）的树突状细胞（dendritic cells，DC）经卡介苗（CGB）活化后诱导的自体淋巴细胞体外抗胰腺癌效应。方法应用rhGM—CSF和rhIL-4从健康人外周血单个核细胞诱导培养DC，用TL负载DC，并用CGB促其成熟。倒置相差显微镜观察DC形态，流式细胞仪检测细胞CD1a、CD83、CD86及HLA—DR表型，ELISA法检测培养上清液中TNF—α和IL-12p 70含量。CCK-8检测DC诱导自体混合淋巴细胞的增殖率以及活化淋巴细胞对PANC1、PaTu8988及SCG7901细胞的杀伤率。结果CGB活化负载TL的DC后，CD83和CD86阳性率分别从（3．7±0．3）％和（38．6±5．0）％显著增加至（16．5±0．6）％和（76．5±2．5）％（P〈0．05）；DC分泌的IL-12p70和TNF—α量分别从（20．18±2．06）pg／ml和（61．38±1．19）pg／ml显著增加至（62．48±3．80）pg／ml和（592．53±17．96）pg／ml（P〈0．01）；DC与自体混合淋巴细胞以1：100、1：50、1：10、1：5的比例共培养，淋巴细胞增殖率分别从（3．90±1．41）％、（4．07±0．40）％、（3．39±1．05）％、（2．82±0．39）％显著增加至（55．38±3．58）％、（75．00±2．54）％、（77．07±3．40）％、（99．07±2．40）％（P〈0．01）；DC活化淋巴细胞对PANCl细胞的杀伤率在效靶比为1：5、1：10、1：20、1：50时分别可达（71．73±0．46）％、（49．44±0．98）％、（31．76±2．77）％、（19．03±3．04）％，但对PaTu8988和SCG7901细胞的杀伤率显著降低。结论经CGB活化的胰腺癌DC疫苗成熟度增加，体外表现出高效特异的抗胰腺癌细胞的效应。Objective To evaluate in vitro anti tumor effect of host lymphocyte primed by Calmette- Guerin bacillus （CGB） activated dendritic cells （DC） based PANC1 lysate. Methods DCs were obtained from peripheral blood mononuclear cells of healthy volunteer and cuitured by rhGM CSF and rhlL 4. DC vaccines for pancreatic cancer were loaded with PANC1 tumor lysate （TL） and were further maturated by CGB. CD1a, CD83, CD86 and HLA-DR phenotype was characterized by flow cytometer, and IL-12p70 and TNF-α concentration in DC culture supernatant were measured by ELISA. Autologous mixed lymphocyte proliferation and the cytotoxicity of cytotoxic T lymphocytes primed by activated DCs to PANC1, PaTu8988 and SCG7901 tumor cells was measured by CCK 8 test. Results When DCs based PANC1 lysate were activated by CGB, the expression rates of CDs3 and CD86 were increased from （ 3.7±0.3 ） % and （ 38.6±5. 0） % to （ 16.5±0.6）% and （76.6 ±2.5）% （ P 〈 0.05）. The concentrations of cytokines ILp12p70 and TNF-α were increased from （ 20.18±2.06 ） pg/ml and （ 61.38±1.19 ） pg/ml to （ 62.48±3.80 ） pg/ml and （ 592.53±17.96）pg/ml （P 〈0.01）. When co-cultured with CGB activated DCs based PANCI lysate in proportion of 1： 100,1：50, 1 ： 10 and 1 ： 5, the proliferation rate of the lymphocytes from （3.90±1.41 ）%, （4.07±0.40）%, （3.39± 1.05）%, （2.82 ±0.39）% significantly increased to （55.38 ± 3.58）%, （75.0 ±2.54） %, （ 77.07± 3.4 ） %, （ 99.07±2.4） % （ P 〈 0.01 ）, respectively. The killing effects of lymphocytes stimulated by DCs at E/T of 1：5, 1： 10, 1：20, 1：50 were （71.73 ±0.46）%, （49.44±0.98）%, （31.76 ±2.77） %, （ 19.03 ± 3.04） % ; but the killing effects on PaTu8988 and SCG7901 were significantly decreased. Conclusions DC vaccines for pancreatic cancer could be more maturated when activated by CGB, and could show a high capability of anti-tumor in vitro.

关 键 词：胰腺肿瘤 树突细胞 卡介苗 

分 类 号：R392[医药卫生—免疫学] R735.9[医药卫生—基础医学]