Loss Of CX3CR1Protect Heptic Granuloma In Schistosomiasis Japonicum Infection And The Mechanism Of Research

From the perspective of immunology, schistosomiasis is, in fact, a kind of participation of cellular and humoral immunity, egg granulomatous disease.Recruitent of macrophages to granuloma mostly contribute to schistosomiasis granuloma development. The precise role of chemokine receptor CX3CR1which mediates both the chemoattractant and adhesion of macrophages is just been elucidated in variety inflammation, but results disparate. In this study, we observed heightened CX3CR1levels in the liver following infection with schistosomiasis japonicum. Here we generated a model in which CX3CR1deficiency mice, produced the following effects when CX3CR1deficiency mouse compared with wild-type mice groups after8weeks in schistosomiasis japonicum infection:Increase the body weight, decreased hepatomegaly, decreased splenomegaly, decreased swelling of the colon, effective protection the function of the liver. Loss of CX3CR1significantly impaired macrophages in granulomatous infiltration, reduced the area of the granuloma. In vitro, produced the following effects when CX3CR1deficiency macrophages compared with wild-type macrophages after soluble egg antigen stimulate96hours: reduced TNF-a, TGF-beta, iNOS content, increased Arg-1and IL-10content, make more to M2macrophages (F4/80+CDllb+CD206+). Mechanistic studies revealed that CX3CR1deficiency macrophages enhanced phosphorylation STAT-6and downstream molecules PPAR-r signaling. These data are not only important for the understanding of the exact role CX3CR1played in schistosome granuloma, but also important for assessing CX3CR1-based therapeutic strategies in clinical settings. Our data show that in CX3CR1knock out mice protection from schistosome granuloma after infection with schistosomiasis japonicum, characterized by the promotion of M2polarization.Objective:Study chemokine receptor CX3CR1, after missing in mice infected with schistosoma japonicum, influence on liver granuloma formation, through the animal and cell experiment and observe the pathological changes of animal specimens and discussed from the view of immunology to elucidate the mechanism of molecular biology.In the future to further follow-up study, evaluation on the basis of CX3CR1, schistosomiasis japonica in the clinical treatment strategy provides an important animal experimental data, the mechanism and treatment of new ideas.Methods:Use6weeks of C57BL/6mice and rat age CX3CR1gene deletion of C57BL/6mice, through the abdominal wall skin infection cercaria method of schistosome infection.8weeks after infection, and using HE staining analysis area of schistosomiasis granuloma, immunofluorescence double dyeing technique is used to analyze qualitatively the macrophages in invasion of surrounding the granuloma.By extracting wild type C57BL/6mice and CX3CR1gene deletion of C57BL/6mice abdominal cavity the original generation of macrophages, and schistosoma japonicum extract antigen of schistosoma japonicum eggs, eggs together after96hours, application method of enzyme linked immunosorbent assay (ELISA) determination of TNF-a, IL4, IL-10, TGF-beta content, protein immunoblot method (Western blot) examined CX3CR1, iNOS, Arg-1, The application of flow cytometry to detect macrophages subtypes M1, M2, surface markers and the proportion of respectively and the STAT-6protein molecules on the macrophage signaling pathway, phosphorylation of STAT-6and downstream protein molecules PPAR-r.Results using mean value plus or minus the standard error deviation (SEM).T test statistical analysis of data.In all cases, are thought to have statistical significance for p<0.05.Results:(1) Loss of CX3CR1mice get a effective protection in the biopsy samples after infected S. japonicum. In the wild type mice and CX3CR1gene deletion mice group with statistical significance (p<0.05).(2) Loss of CX3CR1damaged cell signaling pathways, reduced the eggs deposited parts inflammatory cell infiltration especially the macrophages in the liver, and then, reduced area of the egg granuloma. In the wild type mice and CX3CR1gene deletion mice group with statistical significance (P<0.05).(3) CX3CR1knock out mice protection from schistosome hepatic granuloma after infection with S. japonicum, characterized by the promotion of Th2polarization. In the wild type mice and CX3CR1gene deletion mice group with statistical significance (P<0.05).(4) The macrophage deficiency CX3CR1leads to more expression of the M2subtype protein than WT macrophage. In the wild type mice and CX3CR1gene deletion mice group with statistical significance (P<0.05).(5) Signaling pathway IL-4(CD206)-STAT6-PPAR-r in CX3CR1-/-macrophages upregulate M2macrophage-specific genes programming in the wild type mice and CX3CR1gene deletion mice group with statistical significance (P<0.05).Conclusion:After CX3CR1knock out mice infected with schistosomiasis japonica, can effectively reduce the pathological process of schistosomiasis granuloma, its main mechanism is to reduce the macrophages in the perimeter of the granuloma intrusion and promote inflammation more to M2macrophages of the macrophage polarization. These data not only make us to understand CX3CR1role in Japanese schistosomiasis granuloma formation provides important data, in order to further follow-up study at the same time, the assessment on the basis of CX3CR1, clinical treatment of schistosomiasis japonica, provides an important train of thought on strategy.