摘要
目的探讨分泌人共刺激分子B7—2的重组卡介苗（rBCG）对人外周血单个核细胞（PBMC）IFN-α表达、免疫增强及杀伤膀胱癌细胞的作用。方法SDS—PAGE和ELISA检测rBCG的表达产物hB7—2；rBCG和野生型BCG（wBCG）分别与PBMC共同培养，以单纯PBMC为对照，在不同时段收集培养液上清，经ELISA法检测上清液中IFN—α的表达水平；将BCG激活杀伤细胞（BCG activated kille rcell，BAK细胞）与人膀胱癌EJ细胞共同培养，采用乳酸脱氢酶（LDH）释放试验检测活化免疫细胞对膀胱癌细胞的杀伤作用。结果ELISA法检测出培养上清液hB7—2含量为3．8U／ml。rBCG诱导PBMC产生细胞因子的表达明显高于同浓度的wBCG组（P〈0．05）；rBCG诱导的PBMC抗癌效应高于对照组诱导的抗癌效果（P〈0．05）。结论rBCG诱导人PBMC高表达IFN-α，并通过增强人PBMC细胞的作用来提高抗膀胱肿瘤作用。
Objective To investigate the expression of IFN-α of the peripheral blood monocytes （PBMC） stimulated by bacille Calmette-Guerin （BCG） expressing recombinant human B7-2, and the antitumor effect of BCG activated killer cells（BAK）. Methods Expression of human B7-2 was detected by SDS-PAGE and ELISA. Recombinant BCG and wild-type BCG were used to stimulate PBMC in different concentrations in vitro. Supernatant was collected at various time points and IFN-α was detected by an enzyme-linked immunosorbent assay （ELISA）. MTT assay was used to observe the effects of recombinant BCG on proliferation of T cell, and LDH assay was used to study antitumor cytotoxicity of BAK cells. Results The concentration of human B7-2 in culture was 3.8 U/ml by ELISA. Compared with wild-type BCG, recombinant BCG can induce more IFN-α. The results of the LDH release assay showed that the anti-tumor activity of BAK cells stimulated by recombinant BCG was 2.14 fold higher than that of wild-type BCG. Conclusion The expression of IFN-α in PBMC stimulated by recombinant BCG is higher than that stimulated by wild-type BCG, suggesting that enhanced antitumor activity of BAK when bladder cancer cells could be enchanced by using recombinant BCG.
出处
《肿瘤研究与临床》
CAS
2009年第10期654-656,659,共4页
Cancer Research and Clinic
关键词
膀胱肿瘤
卡介苗
细胞毒性
免疫
Urinary bladder neoplasms
BCG vaccine
Cytotoxicity, immunologic
