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目的在日本血吸虫早期感染模型和SEA致敏模型中,研究IGTP和IRG-47缺失小鼠在抵抗日本血吸虫感染过程中IL-10的免疫调节作用。方法以IGTP基因敲除(IGTP-/-)小鼠、IRG-47基因敲除(IRG-47-/-)小鼠和野生型(WT)小鼠为研究对象,建立日本血吸虫早期感染模型和可溶性虫卵抗原(SEA)致敏模型。在早期感染模型中,以HE染色观察耳廓的病理变化,间接ELISA法检测耳廓皮片培养上清中Th1/Th2型细胞因子的表达水平;在SEA致敏模型中,以间接ELISA法检测血清中日本血吸虫特异性IgG抗体、脾单个核细胞培养上清中Th1/Th2型细胞因子的表达水平;以及流式细胞术检测脾细胞亚群比例的变化。结果日本血吸虫感染7d后,IGTP-/-、IRG-47-/-和WT小鼠的耳廓病理变化无显著差异;但IRG-47-/-小鼠耳廓培养上清中IL-10水平显著低于IGTP-/-小鼠。SEA致敏小鼠3周后,IGTP-/-小鼠血清中SEA特异性IgG抗体水平显著低于WT小鼠;IRG-47-/-小鼠脾细胞在体外SEA刺激下,产生的IL-10水平显著高于IGTP-/-小鼠和WT小鼠;流式结果显示IRG-47-/-小鼠脾中Th2细胞比例显著低于WT小鼠。结论SEA是IRG-47缺失小鼠抵抗日本血吸虫、促使宿主产生保护性应答的重要调节因素,IL-10在此过程中是重要的免疫调节分子。
Objective To investigate the immunomodulatory effects of IL-10 on IGTP and IRG-47-deficient mice against Schistosoma japonicum infection in the early stage of Schistosoma japonicum infection and SEA sensitization. Methods IGTP knockout (IGTP - / -) mice, IRG-47 knockout (IRG-47 - / -) mice and WT mice were used as study objects to establish an early infection model of Schistosoma japonicum Soluble egg antigen (SEA) sensitization model. In the early infection model, the pathological changes of the auricle were observed by HE staining and the expression of Th1 / Th2 cytokines in the culture supernatant of the pinna were detected by indirect ELISA. In the SEA sensitization model, indirect ELISA The serum level of Schistosoma japonicum specific IgG antibody and the expression level of Th1 / Th2 cytokines in the spleen mononuclear cell culture supernatant, and the change of the proportion of splenocyte subsets detected by flow cytometry. Results There was no significant difference in the auricular pathological changes between IGTP - / -, IRG-47 - / - and WT mice 7 days after infection with Schistosoma japonicum; however, the level of IL-10 in the auricle culture supernatant of IRG-47 - / - Significantly lower than IGTP - / - mice. After 3 weeks of SEA sensitized mice, the levels of SEA-specific IgG antibodies in IGTP - / - mice serum were significantly lower than those in WT mice. The splenocytes of IRG-47 - / - mice were stimulated by SEA in vitro, 10 levels were significantly higher than IGTP - / - mice and WT mice; flow cytometry results showed that the proportion of Th2 cells in the spleen of IRG-47 - / - mice was significantly lower than that of WT mice. Conclusion SEA is an important regulator of IRG-47-deficient mice against Schistosoma japonicum, which can induce host’s protective response. IL-10 is an important immunomodulatory molecule in this process.