细胞信号转导抑制剂对日本血吸虫感染小鼠Th1/Th2免疫偏移的调控

REGULATION OF SIGNALING INHIBITORS ON SPECIFIC TH1/TH2 IMMUNE DEVIATION OF MICE INFECTED WITH SCHISTOSOMA JAPONICUM

目的 观察酪氨酸蛋白激酶 (TPK)、蛋白激酶 C(PKC)和磷酯酰肌醇 - 3-激酶 (PI- 3K)特异性抑制剂(Tyrphostin- 2 5、D- sphingosine和 Wortmannin)分别特异性抑制 TPK、PKC和 PI- 3K后 ,对小鼠脾 CD4 + T淋巴细胞经虫卵可溶性抗原 (SEA)诱生 IFN- γ和 IL- 4的表达水平及对 Th1/Th2免疫偏移的影响。 方法 分别于小鼠感染日本血吸虫后 0、4、8和 12周 ,用单克隆抗体分离脾 CD4 + T淋巴细胞体外培养 ,并进行细胞信号转导抑制试验 ,用 EL ISA夹心法和硝酸还原酶法分别测定小鼠脾 CD4 + T淋巴细胞体外培养上清 IFN-γ、IL - 4和一氧化氮 (NO)表达水平。 结果 Tyrphostin- 2 5对 IFN-γ和 IL - 4水平的抑制作用均非常显著 (P<0 .0 1) ,D- sphingosine主要影响 IL - 4的表达 (P<0 .0 1) ,而 Wortmannin则主要影响 IFN-γ的表达 (P<0 .0 1) ,对反映 Th1/Th2免疫平衡的 Th2分化指数分析表明 ,D-sphingosine可使 Th2免疫应答优势减弱 ,而 Wortmannin则可使 Th2免疫应答优势增强。对脾 CD4 + T淋巴细胞培养上清 NO水平检测结果显示 ,NO水平动态与 IFN-γ相一致 ,NO水平主要受 TPK抑制剂 Tyrphostin- 2 5和 PI- 3K抑制剂Wortmaninn的影响 ,结合该两种信号分子抑制剂对 Th1/Th2细胞因子表达水平的作用结果分析 ,

Objective To observe the effects of signaling inhibitors of tyrosine protein kinase(TPK), protein kinaseC(PKC) and phosphatidylinositol 3 kinase(PI 3K) (Tyrphostin 25, D sphingosine and Wortmannin respectively) on the expression of IFN γ and IL 4 and Th1/Th2 immune deviation in mice infected with Schistosoma japonicum . Methods Tyrphostin 25, D sphingosine and Wortmannin was respectively used in mice splenic CD4 + T lymphocyte culture stimulating with solution egg antigen(SEA) at 0?4?8 and 12 weeks after infection. The kits of ELISA and NO were respectively used for the quantitative determination IFN γ, IL 4 and NO in murine splenic CD4 + T lymphocyte culture medium and the effects on Th1/Th2 immune deviation were observed. Results Tyrphostin 25 can significantly inhibit the expression of IFN γ and IL 4( P <0.01), D sphingosine mainly inhibit the expression of IL 4 and Wortmannin mainly inhibit the expression of IFN γ( P <0.01). Th2 differentiation index was used to evaluate the immune regulation balance of Th1/Th2, the results showed that Th2 response was reduced using D sphingosine, and Th2 response was enhanced while Wortmannin was used. The kinetic of expression of nitric oxide(NO) were correspondence with IFN γ and the expression of NO were mainly inhibited by Tyrphostin 25 and Wortmannin. These data further demonstrated above the mechanism. Conclusion These findings suggest that signaling inhibitors can modulate Th1/Th2 immune deviation and further elucidate the mechanism of regulation of egg granuloma formation of S. japonicum . It may represent a novel potential therapeutic approach for controlling egg granuloma formation of S. japonicum.