CAJ | 학술논문

目的检测人外周血γδT细胞的信号转导分子ζ链相关蛋白-70 (ZAP-70)的表达.方法应用淋巴细胞分离液常规分离获取健康人外周血单个核细胞(PBMC),用结核杆菌低分子多肽抗原(Mtb-Ag)刺激PBMC增殖培养,10d后收集细胞,用免疫磁珠阳性分选法分离获取高纯度的γδT细胞；采用免疫印迹方法检测γδT细胞内的ZAP-70分子.结果新鲜分离的PBMC中γδT细胞的比例仅为4.9％,Mtb-Ag刺激培养10d后升为69.2％,免疫磁珠阳性分选后达99.3％.免疫印迹显示检测到γδT细胞内相对分子质量为70 000的ZAP-70分子的表达.结论 ZAP-70分子在活化增殖的人外周血γδT细胞内高表达.
목적 검측인외주혈γδT세포적신호전도분자ζ련상관단백-70 (ZAP-70)적표체.방법 응용림파세포분리액상규분리획취건강인외주혈단개핵세포(PBMC),용결핵간균저분자다태항원(Mtb-Ag)자격PBMC증식배양,10d후수집세포,용면역자주양성분선법분리획취고순도적γδT세포；채용면역인적방법검측γδT세포내적ZAP-70분자.결과 신선분리적PBMC중γδT세포적비례부위4.9％,Mtb-Ag자격배양10d후승위69.2％,면역자주양성분선후체99.3％.면역인적현시검측도γδT세포내상대분자질량위70 000적ZAP-70분자적표체.결론 ZAP-70분자재활화증식적인외주혈γδT세포내고표체.
Objective To detect the expression of signal transduction molecule ζ chain-associated protein 70 (ZAP-70) in proliferated γδT cells derived from human peripheral blood.Methods The peripheral blood mononuclear cells (PBMCs) were conventional separated by using lymphocyte separation liquid,and obtained from healthy volunteers.PBMCs were stimulated by low-molecular peptide antigen of mycobacterium tuberculosis (Mtb-Ag).Following incubation for 10 days,the T cells were harvested for purification of γδT cells by using the immunomagnetic beads selection algorithm.The expression of ZAP-70 in γδT cells was detected by Western blotting.Results The proportion of γδT cells were 4.9％ in fresh PBMCs,69.2％ at 10 days following incubation with Mtb-Ag,and 99.3％ after immunomagnetic beads selection.ZAP-70 (relative molecular mass of 70 000) expression in γδT cells was detected by Western blotting.Conclusion ZAP-70 is highly expressed in γδT cells following proliferation and activation.