The Study On Dynamic Changes And Immune Function Of Different Types Of Macrophages In Mice Infected With Echinococcus Multilocularis

Objective:To study the dynamic changes of monocytes,macrophages and their related factors in animal models of persistent infection of Echinococcus multilocularis,Analysis of its immune function in persistent infection of Echinococcus multilocularis,Explore the possible links between them and persistent infection with Echinococcus multilocularis.Method:80 female BALB/c mice were randomly divided into normal control group and experimental group by randomization method.The experimental group established a model of infection of the Echinococcus multilocularis by intraperitoneal injection of the original Solution of E.multilocularis.The mice in the control group were intraperitoneally injected with the same amount of normal saline.Collect the required specimens 2 days,8 days,30 days,90 days,180 days after infection.Observing the pathological changes of mouse liver by HE staining and establishing a table of the degree of infection score.Proportion Changes of Ly6C^hi and Ly6C^lo type monocytes in peripheral blood of mice by flow cytometry.Distribution and expression of chemokines and their receptors CX3CL1,CX3CR1,M1 macrophage-associated factor i NOS,M2 macrophage-specific molecule CD163 and monocyte-specific molecule CD11b in mouse liver by immunohistochemistry.The m RNA expression levels of M1 macrophage-associated factor TNF-αand M2 macrophage-associated factor Fizz1 in mouse liver were detected by real-time quantitative PCR.The expression levels of IFN-γ,IL-17,IL-4 and IL-10 in peripheral blood of mice were detected by ELISA.Results:1)HE staining results show that,in the early stage of Echinococcus multilocularis infection,only a few focal necrosis were observed in the liver of the experimental group,and slight inflammatory cell aggregation was observed accidentally around the portal area.In the middle stage of infection,fusion or even bridging necrosis can be seen in the hepatic lobules,and the inflammatory cells around the portal area are obviously increased,and the lesions wrapped by the wall of the Collagen fibers can be seen.In the late stage of infection,the structure of the lobule around the lesion is not obvious,and a large number of inflammatory cells are aggregated.The results of the infection degree score table showed that as the infection time prolonged,the total score showed an overall upward trend.2)The results of flow cytometry show that,compared with the control group,the proportion of Ly6C^hi type monocytes in the peripheral blood of the experimental group began to increase in the early stage of infection,but its proportion gradually decreased with the prolongation of infection time.The proportion of Ly6C^lo type monocytes began to decline significantly after a brief increase in early infection,but began to increase significantly until 90 days of infection,reaching a peak at 180 days of infection.3)The results of immunohistochemistry show that,in the process of infection of Echinococcus multilocularis,compared with the control group,the expression of CX3CL1 in the liver of the experimental group was mainly concentrated in the inflammatory foci and around the lesion.In addition,the expression level begins to increase in the early stage of infection,peaks in the middle stage,and declines in the late stage.The expression level of CX3CR1 in the liver of the experimental group was significantly increased in each infection period,and the overall trend was to increase,decrease,increase,and then decrease.The expression site is scattered in the early but concentrated in the late inflammatory sites.The expression levels of i NOS in the liver of the experimental group gradually rises and peaks in the early and middle stages of infection,but decreases in the late stage.The expression levels of CD163 in the liver of the experimental group increased with the prolongation of infection time,and reached the peak until the late infection.The expression of CD11b in the liver tissue of the experimental group was mainly concentrated in infiltrating monocytes.The expression levels of CD11b in the liver of the experimental group increased with the prolongation of infection time,and reached the peak until the late infection.4)The results of real-time fluorogentic quantitative PCR show that,compared with the control group,the expression level of TNF-αm RNA in the liver of the experimental group began to increase in the early and peaked in the middle stages,and then began to decrease in the late stage.The expression level of Fizz1 m RNA is continuously increased with the prolonged time of infection and peaked until late infection.5)ELISA test results show that,the cytokine of Th1type reaction increases in the early and middle stages of infection,but begins to decrease in the late stage of infection.The cytokine of Th2 type response did not increase significantly in the early stage of infection,but increased significantly in the middle stage,although decreased in the late stage but still higher than the normal control group.Conclusion:Monocytes in peripheral blood differentiate from early Ly6C^hi to late Ly6C^lo.Its amount recruited in liver tissue increases with increasing chemokines and their receptors.Indirectly indicating that macrophages of two different sources play a role in the infection of Echinococcus multilocularis.In the early stage of infection,both of them may be involved in the infection process of Echinococcus multilocularis through M1 type macrophage mediated Th1 immunity.In the late stage,the M2 type may be the main differentiation direction of them,and involved in the infection of Echinococcus multilocularis by Th2 type immunity.