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[Imbalance of Th17/Tregs promotes egg granuloma formation of liver with Schistosomiasis japonicum in mice].

Objectives To investigate the effect of the imbalance of Th17/Treg on egg granuloma formation of liver with Schistosomiasis japonicum. Methods The BALB/c mice were infected with Schistosoma japonicum cercariae to establish a model of Schistosomiasis japonica. The blood samples, liver tissues and spleen tissue were harvested at the 2nd, 4th, 6th, 8th week, respectively. HE staining and Masson staining were performed to assess the pathological characteristics of the liver. Flow cytometry (FCM) was conducted to evaluate the proportion of CD4+ T cell subsets including Th17 cells and Tregs in liver and spleen tissue. The quantitative real-time PCR (qRT-PCR) was carried out to investigate the mRNA level of cytokines including RORγt, FOXP3, IL-6, IL-17, IL-23 and IL-10 in liver tissues. Finally, ELISA was performed to assess the serum level of cytokines including IL-6, IL-17, IL-23 and TGF-β. Schistosoma japonicium soluble egg antigen (SjSEA) were prepared to stimulate mouse spleen cells in vitro. qRT-PCR was carried out to investigate the mRNA level of cytokine including RORγt and FOXP3 and ELISA was performed to assess the expression level of cytokines including IL-6, IL-17, IL-23 and TGF-β at different time points. Results HE and Masson staining demonstrated that inflammatory cell infiltration, schistosome egg granuloma formation and the collagen deposition increased in the liver tissue after the 4th week. The longer the infection, the more severe the liver pathology. In the liver and spleen tissues, the percentage of Th17 cells of infection group (2nd, 4th and 6th weeks) were significantly higher than the healthy group. The percentage of Tregs in the liver tissues of infection group (4th, 6th and 8th weeks) were significantly higher than the healthy group, and the percentage of Tregs in the spleen of infection group (2nd and 4th weeks) were significantly higher than the healthy group. Th17/Treg ratios in the liver of infection group were lower than the healthy group. Th17/Treg ratios in the spleen of infection group (2nd and 4th weeks) were lower than the healthy group, while it increased in the 6th week. At the same time, the levels of Th17 cells and Tregs related nuclear transcription factors and cytokines showed similar dynamic changes as the percentages of T cell subsets. SjSEA can induce the differentiation of Th17 and Tregs and the expression of related cytokines and transcription factors. Conclusion Th17 cells may play a major role in liver pathology, and the imbalance of Th17 cells/Tregs was closely related to the schistosome egg granuloma formation.