Fig 1.
Quantitative changes in splenic Treg and Th17 cell percentages and the relationship between Treg and Th17 subsets.
Single-cell spleen suspensions from C. sinensis-infected and control mice (NC) were used to analyze Th17 and CD4+CD25+Foxp3+regulatory T cell percentages among CD4+ T cells by flow cytometry (A). Representative blots of IL-17+ and CD25+Foxp3+cells in splenic CD4+cell populations. B and C show quantitative changes in splenic Treg and Th17 cell percentages in CD4+T cells. The Treg/Th17 ratio was calculated (D). Data are expressed as mean ± S.E.M. Asterisks indicate statistically significant differences between infected and NC mice, **P<0.01, ***P<0.001, compared with normal control mice.
Fig 2.
Hepatic expression levels of Foxp3 and RORγt in the mice infected by Clonorchis sinenis.
Liver sections from C. sinesis-infected and NC mice were homogenized; and the expression levels of RORγt (A) and Foxp3 (B) were determined by western-blot. Data are expressed as mean ± S.E.M. Asterisks indicate statistically significant differences between infected and NC mice; **P<0.01, compared with normal control mice.
Fig 3.
Kinetics of serum IL-17A, IL-6, IL-2, and TGF-β1.
Serum from each mouse was screened with cytometric bead array (CBA) to detect IL-6 (A), IL-17A (B), and IL-2 (C); enzyme-linked immunosorbent assays (ELISA) was used to measured TGF-β1 (D). Data are expressed as mean ± S.E.M. Asterisks indicate statistically significant differences between infected mice and NC mice at a particular time point, *P<0.05, **P<0.01, and ***P<0.001, compared with normal control mice.
Fig 4.
Representative histology of liver sections stained with hematoxylin and eosin (H&E).
A and a show liver sections from NC mice. In infected mice, massive inflammatory cells were infiltrated around the portal areas accompanied by cholangiocyte proliferation on day 14 post-infection (PI). (B and b). The numbers of inflammatory cells decreased, but bile duct hyperplasia was exaggerated on day 28 PI. (C and c) and 56 PI. (D and d). Capitals and lowercase letters indicate magnifications of ×100 and ×400 times respectively.
Fig 5.
Collagen depositions in the liver of Clonorchis sinensis-infected BALB/c mice.
Compared to NC mice (A), collagen fibers were slightly stained blue by Masson’s trichrome staining on day 14 post-infection (PI) (B), and the degree of staining were increased on day 28 PI (C) and 56 PI (D). Correlations were calculated between the degree of Treg/Th17 imbalance and liver fibrosis caused by C. sinensis (E).