PHA与PMA对外周血单个核细胞产生IL-4、IFN-γ水平的研究

Comparison of phytohemagglutinin and phorbol myristate acetate + ionomycin stimulative effect on peripheral blood mononuclear cells

目的 :比较 PHA(植物血凝素 )与 PMA(佛波醇乙酯 )对 PBMCs(外周血单个核细胞 )的刺激效果。方法 :选择健康人群 ,取外周静脉血 ,分离得到 PBMCs,分别加入 PHA+ ionomycin(离子霉素 )及 PMA+ ionomycin进行刺激并孵育 2 h、4h、6h、12 h、2 4h、48h,采用双抗体夹心 ABC-ELISA法 ,分别测定不同时相培养上清中 IL -4和 IFN-γ水平。结果 :刺激 2 h后 ,PHA组与 PMA组 IFN-γ和 IL -4水平无差别 ;刺激 4h后 ,PMA组 IFN-γ和 IL-4水平均高于 PHA组 (P<0 .0 1) ;刺激 6h后 ,PHA组 IFN-γ水平低于 PMA组 (P<0 .0 5 ) ,IL -4水平高于 PMA组 (P<0 .0 5 ) ;刺激 12 h后 ,PHA组 IFN-γ与 PMA组接近 ,两组 IL -4水平亦无差别 ;刺激 2 4h和 48h后 ,PHA组 IFN-γ和 IL-4水平均高于 PMA组 (P<0 .0 5 )。结论 :PHA和 PMA对 PBMCs不同时间的刺激效果不同。

Objective:To compare stimulative effect of phytohemagglutinin(PHA)+ionomycin and phorbol myristate acetate(PMA) +ionomycin on peripheral blood mononuclear cells(PBMCs).Methods:Choosing healthy persons,taking and separating peripheral venous blood to gain PBMCs.Adding PHA+ionomycin and PMA+ionomycin respectively to stimulate and incubate,with the method of enzyme-linked immunosorbant assay(ELISA) to measure the level of IL-4 and IFN-γ in the serum.Results:After 2 hours of stimulation, the level of IFN-γ and IL-4 in both groups is similar. After 4 hours of stimulation, the level of IFN-γ and IL-4 in PMA group is both higher than that in PHA group(P<0.01).After 6 hours of stimulation, the level of IFN-γ in PHA group is lower than that in PMA group(P<0.05),the level of IL-4 in PHA group is higher than that in PMA group(P<0.05).After 12 hours of stimulation, the level of IFN-γ in PHA group is close to that in PMA group,the level of IL-4 in both groups is similar.After 24 hours and 48 hours of stimulation,the level of IFN-γ and IL-4 in PHA group is both higher than that in PMA group(P<0.05).Conclusion:Stimulative effect of PHA and PMA on peripheral blood mononuclear cell is different due to different stimulation times.