【摘要】 目的研究肺炎链球菌热休克蛋白40(heat shock protein 40,HSP40)诱导巨噬细胞免疫应答的机制。方法表达重组HSP40蛋白,并在体外诱导培养小鼠骨髓来源巨噬细胞(bone marrow derived macrophage,BMDM);使用NF-κB、PI3K和JAK的抑制剂预处理BMDM后,ELISA检测其对HSP40诱导的TNF-α和IL-6表达的影响;HSP40刺激BMDM后,Western blot法检测NF-κBp65和Akt的磷酸化水平。结果 NF-κB和PI3K抑制剂可显著下调IL-6和TNF-α的表达,而JAK抑制剂无此效果。HSP40可增强NF-κBp65和Akt磷酸化水平。结论 NF-κB和PI3K-Akt通路参与调控肺炎链球菌HSP40诱导BMDM免疫应答的过程。更多还原

【Abstract】 Pneumococcal HSP40 was reported to be an effective candidate vaccine,and we have found that HSP40 could induce immune response in mouse bone marrow derived macrophage(BMDM) though p38 MAPK and JNK signaling pathways.In this study,we investigated the roles of NF-κB,PI3 K and JAK pathways in the course of immune response in macrophage induced by HSP40.Recombinant HSP40 protein was expressed and BMDM was cultured successfully in vitro.The effects of the pretreatment of NF-kB,PI3 K and JAK inhibitors on the secretion of TNF-α and IL-6 induced by HSP40 were detected by ELISA;the phosphorylation levels of NF-κBp65 and Akt in BMDM were measured by Western blotting after HSP40 stimulation.Data showed that the inhibitors of NF-κB and PI3 K could significantly down-regulate the expressions of IL-6 and TNF-α,but the inhibitor of JAK did not display such effect.HSP40 could enhance the phosphorylation levels of NF-κBp65 and Akt.These results indicate that NF-κB and PI3K-Akt signaling pathways are involved in regulating the immune response of BMDM induced by Pneumococcal HSP40.更多还原