The Role Of Tfh Cell And Related Molecules In Echinococcus Multilocularis Infection

Objective:To study Tfh cells and transcription factor Bcl-6 related cytokines CXCR5,ICOS Echinococcus multilocularis(Em)Changes infection in Balb/c mice to explore the role of infection in alveolar hydatid.Methods:60 female BALB/c mice were randomly divided into a control group and the experimental group and the control group by intraperitoneal injection of saline,the experimental group were inoculated intraperitoneally with the former head larva preparation of alveolar hydatid infection mouse model.Were infection 2,8,30,90,180 and 300 days of each 5 mice were sacrificed,and the liver tissue and spleen tissue,the pathological changes of the liver tissue was observed by HE staining;Through immunohistochemical detection of mice infected with alveolar hydatid liver and spleen of CXCR5,ICOS,Bcl-6 expression site and distribution;Take the liver tissue,alveolar hydatid infection related phenotypes CXCR-5,ICOS and transcription factor Bcl-6 detection,to analyze the expression level of the mouse liver by real-time quantitative PCR(QRT-PCR);Organization of the spleen and lymph nodes,lymph nodes and spleen cells of infected mice by flow cytometry alveolar hydatid Tfh cell subsets in;Serum,alveolar hydatid infected mouse serum cytokines IL-4 and IL-10 levels by ELISA method by CBA method detection level of the serum of mice infected with alveolar hydatid antibody IgGl subclasses;Results:1)alveolar hydatid mid-infected mice visible obvious alveolar hydatid vesicles late vesicle Center necrosis and difficult to peel and the surrounding tissue,intra-abdominal adhesions;2)HE staining under light microscope observations:early liver tissue structure in the experimental group of mice infected with alveolar hydatid showed no abnormal change liver cell boundary is clear and neat;The infection medium-term visible to a large number of inflammatory cell infiltration,bile duct hyperplasia,liver library whether cell proliferation and focal necrosis of liver cells;advanced stage of infection can be seen around the portal area of liver diffuse fibrous proliferation,fibrosis,and fibrous tissue in the number of liver cells surrounding;3)immunohistochemistry results showed:alveolar hydatid infection early,CXCR5,ICOS and BCL-6 was not expressed or expressed at low levels;The alveolar hydatid infection in late,with the increase in the liver of mice alveolar hydatid increase,CXCR5,ICOS and BCL-6 expression were significantly increased(P<0.05);4)QRT-PCR show display compared with the control group:In the early stages of infection ICOS mRNA and CXCR5 mRNA are expressed at low levels,infection interim ICOS mRNA expression was significantly increased(P<0.05),and advanced stage of infection continued to maintain a high level;CXCR5 mRNA expression in infected mid-significantly increased(P<0.05),and continued to maintain a high level in the late stage of infection;5)flow cytometry results showed that compared with the control group:the mouse lymph Tfh cells,Tfh proportion of cells in the infected late significantly increased(P<0.05);mouse spleen Tfh cells,Tfh cell ratio infection in late increased significantly;6)the CBA test results show that compared with the control group:infected the early mouse serum IL-4 levels without significant changes began to increase infection of the mid-level of IL-4,then to maintain a higher level(P<0.05);7)ELISA test results show that compared with the control group:the level of infection of early mouse serum IgG1 maintained at 0.64pg/ml infection interim IgG1 levels began to increase,and then to maintain a high level(P<0.05).Conclusion:Tfh cells in alveolar hydatid infection as early as in the late and peripheral lymphoid organs change characteristics of alveolar hydatid infection can cause changes in mouse liver fibrosis,aggravate and prolong the course of time;The early stages of infection,alveolar hydatid liver tissue structure no obvious abnormal changes Tfh cells and their cytokine CXCR5,ICOS,BCL-6,IL-4,also no abnormal changes in serum IgG1 levels maintained in 0.64pg/ml.Alveolar hydatid infection mid mouse liver,spleen tissue Tfh cell ratio were increased the Tfh cell factor CXCR5,ICOS,BCL-6,IL-4 were significantly higher proportion of alveolar hydatid infection may be involved in the development;Alveolar hydatid advanced stage of infection,mouse liver,spleen tissue and lymph nodes Tfh cells increased the proportion of Tfh cytokines CXCR5,ICOS,BCL-6,IL-4 increased the proportion of jointly participate in the development of persistent alveolar hydatid infection;In short,the the Tfh cell and the transcription factor Bcl-6,CXCR5,ICOS increase in the amount of expression of related cytokines may promote the development of the persistent occurrence of alveolar hydatid infection.