重组金黄色葡萄球菌肠毒素C3生物学活性的初步研究  

Expression and preliminary bioactivity analysis of the recombinant Staphylococcus aureus enterotoxin C3

作　　者：李先平[1] 王敏[1] 武婷[2] 梅城 曹虹[1] 

机构地区：[1]中南大学湘雅二医院检验科,长沙410011 [2]中南大学湘雅医学院医学检验系2006级,长沙410013

出　　处：《中国免疫学杂志》2012年第7期589-593,共5页Chinese Journal of Immunology

基　　金：湖南省自然科学基金(06JJ4051)

摘　　要：目的:表达和纯化重组金黄色葡萄球菌肠毒素C3(rSEC3),并初步探讨rSEC3的生物学活性。方法:将转化有pET-32a(+)-SEC3重组质粒的大肠杆菌BL21经IPTG诱导表达、纯化后,MTT法检测0.1、0.5、1、5、10、50、100μg/ml的rSEC3促进人外周血单个核细胞(PBMC)增殖及抑制人宫颈癌细胞(Hela细胞)、人食管癌细胞(KYSE细胞)的作用。结果:重组金黄色葡萄球菌肠毒素C3在大肠杆菌BL21中成功表达。与空白对照相比,0.1～100μg/ml的rSEC3对PBMC均有显著的促增殖作用(P<0.05),1μg/ml时促增殖作用最强(t=113.851,P=0.000),和100μg/ml PHA作用相似(t=0.693,P=0.527)。以Hela细胞为靶细胞,培养48小时后,与空白对照相比,0.5、1、5μg/ml的rSEC3能显著增强PBMC对Hela细胞杀伤作用(P<0.05),抑瘤率分别为(70.44±9.27)%、(93.65±8.05)%、(103.40±6.65)%;以KYSE细胞为靶细胞,与空白对照相比,3个浓度的rSEC3也都有抑瘤作用(P<0.05),抑瘤率分别为(28.10±2.72)%、(46.62±4.17)%、(19.35±3.05)%。结论:rSEC3蛋白具有良好促人外周血单个核细胞增殖活性,并能够增强PBMC对肿瘤细胞的杀伤活性。Objective：To express the recombinant Staphylococcus aureus enterotoxin C3 （rSEC3） and preliminary investigate its activity. Methods：E. coli BL21 containing the recombinant plasmids pET-32a（ ＋ ）-SEC3 was expressed and purified. Then the human PBMCs was stimulated with different concentration of rSEC3 at 0.1,0.5,1,5,10,50,100 ug/ml and the promoting proliferation activity was detected by MTT method. Last, the tumor-cytotoxicity effects of Hela and KYSE were examined after being induced 48 h. Results ：The recombinant Staphylococcus aureus enterotoxin C3 was expressed efficiently in E. coli BL21. Compared with blank con- trol, 0.1-100 ug/ml of rSEC3 could significantly promote proliferation of human PBMCs （ P 〈 0.05 ）. 1 ug/ml of rSEC3 was the most efficient dose （ t = 113.851 ,P = 0. 000） and showed a similar effect with 100 ug/ml PHA （ t = 0. 693, P = 0. 527）. After cultured 48 h, rSEC3 at 0.5, 1, 5 ug/ml had a strong depressant effect on Hela by activating PBMC compared with blank control （ P 〈 0.05 ）. The inhibition rates were （ 70.44 ± 9.27 ） %, （93.65 ± 8.05 ） %, and （ 103.40 ± 6.65 ） %. Furthermore, the rSEC3 also enhanced the killing activity to KYSE cells （ P 〈 0.05 ）, and the inhibition rates were （ 28. 10 ± 2.72 ） %, （ 46.62 ± 4. 17 ） %, and （ 19.35 ±3.05 ）% , respectively. Conclusion：The recombinant protein rSEC3 has activity on human PBMCs proliferation, and it can enhance PBMCs killing effect on tumor cells.

关 键 词：金黄色葡萄球菌 肠毒素 表达 生物学活性 

分 类 号：R392[医药卫生—免疫学]