IL-10CD5CD1d、IFN-γTh1及M1型巨噬细胞在多房棘球蚴感染SD大鼠后的免疫调节初步分析

Preliminary immunomodulatory analysis on IL-10CD5CD1d, IFN-γ Th1 and M1 macrophages after SD rat infected by the echinococcus multilocularis

目的初步分析IL-10CD5CD1d、IFN-γTh1、M1型巨噬细胞在多房棘球蚴感染SD大鼠后的免疫调节机制。方法采用STRING及Cytoscape 3.7.1软件通过生信分析预测大鼠IFN-γ、IL-10、CD4、CD5、CD80、CD1d免疫表型之间的联系,将实验分为对照组和模型组,模型组6只SD大鼠通过肝原位接种1 000个活的多房棘球蚴感染SD大鼠,对照组6只,不做任何处理。3个月后利用流式细胞术及ELISA分析免疫细胞表型及炎症因子。结果生信分析发现IFN-γ、IL-10、CD4、CD5、CD80、CD1d之间相互作用紧密,尤其是IL-10、CD5及CD80发挥了主要作用,同时流式结果显示模型组IL-10CD5CD1d比正常对照组显著降低(t=6.164,P=0.000 106),差异有统计学意义,提示模型组CD5CD1d产生的IL-10减少,对Th1及M1型巨噬细胞的作用减弱;CD4^(+)IFN-γ^(+)(t=2.558,P=0.028 462)及CD80;显著升高(t=3.293,P=0.008 110),差异有统计学意义,提示Th1细胞产生的IFN-γ升高,M1型巨噬细胞被激活;同时ELISA结果显示模型组肝组织中IL-10含量降低(t=4.169,P=0.001 920)、IFN-γ含量升高(t=5.187,P=0.000 409),差异有统计学意义,提示IL-10及IFN-γ整体水平与流式检测的结果一致。结论 CD5CD1d在SD大鼠感染多房棘球蚴后IL-10的分泌可能受到抑制,从而导致IFN-γTh1的分泌及M1型巨噬细胞极化。

Objective To primarily analyze the immunomodulatory mechanism of IL-10 CD5 CD1 d, IFN-γTh1 and M1 macrophages after SD rats infected by the echinococcus multilocularis.Methods The connections among the immunophenotypes of IFN-γ,IL-10,CD4,CD5,CD80 and CD1 d were predicted by STRING and Cytoscape 3.7.1 software by means of bioinformatics analysis, and the experiment was divided into the model group and control group.The 6 SD rats in the model group, were injected by orthotopic liver inoculation of 1 000 live echinococcus multilocularis.The 6 SD rats in the control group, weren’t subject to any treatment.The flow cytometry and ELISA were used to analyze immunophenotype and the inflammatory factors three months later.Results The bioinformatics analysis revealed there were closely relations between IFN-γ,IL-10,CD4,CD5,CD80 and CD1 d, especially IL-10,CD5 and CD80,and played an important roles, the flow cytometry showed that IL-10 CD5 CD1 d in the model group was lower than that in the control group(t=6.164,P=0.000 106),and the difference was statistically significant, indicating that IL-10 produced CD5 CD1 d was decreased to weak the effect of Th1 and M1 macrophage;both CD4^(+)IFN-γ^(+)(t=2.558,P=0.028 462) and CD80;significantly increased(t=3.293,P=0.008 110),and the difference was statistically significant, indicating that IFN-γ produced by Th1 increased and M1 macrophage was activated.ELISA showed that the level of IL-10 significantly decreased(t=4.169,P=0.001 920) and IFN-γ increased(t=5.187,P=0.000 409) in model group, and the difference was statistically significant, indicating that the overall level of IL-10 and IFN-γ matched the results of flow cytometry.Conclusion The secretion of IL-10 may be inhibited for CD5 CD1 d when SD rats are infected with echinococcus multilocularis,thereby causing the secretion of IFN-γTh1 and polarization of M1 macrophages.