金黄色葡萄球菌PV杀白细胞素S组分体外诱导单核白血病细胞株THP-1巨噬细胞的研究

Study on the induction of mononuclearleukemia cell THP-1 drived macrophages by S component of Panton-Valentine leukocidin in vitro

目的目前关于金黄色葡萄球菌PV杀白细胞素S组分(LukS-PV)体外诱导人源单核白血病细胞株THP-1巨噬细胞研究较少。文中主要探讨LukS-PV能否体外诱导人源THP-1巨噬细胞极化。方法佛波酯体外诱导THP-1为THP-1巨噬细胞,THP-1经佛波酯刺激48 h、72 h后,流式检测细胞表面标志物CD11b、CD14。采用0.1、0.2、0.4μmol/L的LukS-PV作用THP-1巨噬细胞,分别为0.1μmol/LLukS-PV组、0.2μmol/LLukS-PV组和0.4μmol/LLukS-PV组,另采用佛波酯诱导THP-1巨噬细胞为对照组。流式细胞术检测各组24 h、48 h CD80、CD206的表达,qRT-PCR检测细胞IL-12、TNF-α、TGF-β的mRNA表达水平。结果0.1μmol/LLukS-PV组、0.2μmol/LLukS-PV组、0.4μmol/LLukS-PV组细胞表面CD80表达量较对照组明显增高(P<0.01),0.2μmol/L LukS-PV组表达量最高(P<0.01)。0.2μmol/L LukS-PV组24h IL-12、TNF-αmRNA表达水平(7.32±0.91、5.29±0.51)较对照组(0.71±0.11、0.73±0.14)明显升高(P<0.01),48 h时IL-12、TNF-αmRNA表达水平(11.16±0.78、6.70±0.68)较对照组亦明显升高(P<0.01)。0.2μmol/L LukS-PV组24h、48h细胞表面CD80的表达较对照组明显升高(P<0.000)。结论LukS-PV能够刺激人源THP-1巨噬细胞向M1极化。

Objective There are few studies on the in vitro induction of human leukemia cell line THP-1 macrophages by Staphylococcus aureus PV leukin S component(LukS-PV).This study aims to investigate whether LukS-PV could induce the polariza-tion of Human THP-1 derived macrophage in vitro.Methods THP-1 monocytes were induced to macrophages by Phorbol myristate acetate(PMA).The cell surface markers CD11 b and CD14 were detected by flow cytometry at 48 h and 72 h after the induction,respec-tively.Using 0.1,0.2 and 0.4μmol/LlukS-PV treated THP-1 macrophages as the 0.1μmol/LLukS-PV group,0.2μmol/LLukS-PV group,and 0.4μmol/LLukS-PV group,respectively.The THP-1 driven macrophage induced by PMA was used as the control group.Flow cytometry was used to detect the expression of CD80 and CD206 in each group at 24 h and 48 h.The mRNA expression levels of IL-12,TNF-αand TGF-βwere detected by real-time fluorescence-based quantitative PCR(qRT-PCR).Results The expression of CD80 on the cell surface in the 0.1μmol/LLukS-PV group,0.2μmol/LLukS-PV group and 0.4μmol/LLukS-PV significantly increased(P<0.01),with the highest expression in the 0.2μM LukS-PV group(P<0.01),when comparing to that of the control group.The mRNA expressions of IL-12 and TNF-αin the 0.2μM LukS-PV group significantly increased(7.32±0.91 and 5.29±0.51,respective-ly)at 24 h(P<0.01),and raised in 0.2μM LukS-PV group(11.16±0.78、6.70±0.68)at 48 h(P<0.01),when comparing to those of control group(0.71±0.11 and 0.73±0.14,respectively).Compared with the control group,expression of CD80 on cell surface in the0.2μM LukS-PV group significantly increased at 24 h and 48 h(P<0.000).Conclusion LukS-PV could stimulate the polarization of human THP-1 macrophages toward M1.