重组BCG的抑瘤活性及免疫学机制研究  

Antitumor activity and immunological mechanisms of recombinant BCG

作　　者：薛庆节[1] 李秀真[1] 李运清[1] 杨媛媛[1] 胡文洁[1] 章洪华[1] 吕厚东[1] 李士根[1] 

机构地区：[1]济宁医学院病原生物学教研室,医学免疫学省级重点学科,济宁272067

出　　处：《中国免疫学杂志》2015年第8期1060-1065,共6页Chinese Journal of Immunology

基　　金：山东省自然科学基金项目(ZR2012HM037);山东省高等学校科技计划项目(No.J12LK56,J11LF03);教育部留学回国人员科研启动基金资助项目(20101174)

摘　　要：目的:对重组GM-CSF和EB病毒LMP2A融合基因的BCG(卡介苗)进行抑瘤活性及免疫学机制研究。方法:建立EB病毒阳性肿瘤动物模型,对小鼠成瘤时间、存活情况、肿瘤重量进行分析,检测重组BCG的抑瘤活性;用ELISA方法对重组BCG刺激机体产生的特异性抗体进行检测,乳酸脱氢酶法检测特异性CTL杀伤效应,ELISPOT法检测IFN-γ的分泌水平,流式细胞术、HE染色检测重组BCG免疫小鼠的肿瘤组织中的淋巴细胞浸润情况。用统计学方法对重组BCG免疫效果进行初步分析和评价。结果:肿瘤成瘤时间与其他对照相比明显延迟,肿瘤生长明显缓慢,小鼠生存期延长。ELISA检测结果表明,重组BCG免疫后能产生针对GM-CSF和LMP2A的特异性Ig G抗体,重组BCG免疫组的小鼠能检测到特异性CTL活性,用ELISPOT法检测到免疫小鼠淋巴细胞有IFN-γ分泌;流式细胞术及形态学观察的方法检测到重组BCG免疫的小鼠肿瘤组织中有淋巴细胞的浸润生长。结论:重组BCG诱导C57BL/6小鼠机体产生了体液免疫和细胞免疫反应,重组BCG对EB病毒阳性肿瘤细胞具有明显抑制作用。Objective： To study the antitumor activity and immunological mechanisms of r BCG including GM-CSF and EB virus LMP2 A gene fusion. Methods： Animal models of EB virus-positive tumors was built. The formation time of tumors in mice,survival time,tumor weight was analyzed to detect r BCG anti-tumor activity; ELISA method was used to detect the specific antibodies which was produced in the mice stimulated by r BCG,specific CTL killing effect was detected by lactic dehydrogenase assay,ELISPOT was used to assay the secretion of IFN-γ and flow cytometry,HE staining of tumor tissue was used to detected lymphocyte infiltration in mice immunized with recombinant BCG. Statistical methods were used for r BCG immunization effect preliminary analysis and evaluation. Results：Comparing to other control,tumor formation time was significantly delayed and tumor growth was slow,survival time of mice prolonged. ELISA test results showed that the r BCG immunization groups of mice could produce specific Ig G antibodies of GM-CSF and LMP2 A. Specific CTL activity was detected in mice immunized with r BCG. IFN-γ secretion was detected by ELISPOT method,flow cytometry and morphological observation detected tumor tissue infiltration of lymphocytes growth in mice immunized with r BCG. Conclusion： The r BCG induced a humoral and cellular immune responses and induced C57 BL /6 mice to produce a strong antitumor effect and the EB virus-positive tumor cells was significantly inhibited.

关 键 词：重组BCG 抗肿瘤 细胞免疫 EB病毒 

分 类 号：R755[医药卫生—皮肤病学与性病学]