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. 2014 May 1;10(5):e1004097. doi: 10.1371/journal.ppat.1004097

Figure 2. Tfh cells promote liver pathology.

Figure 2

(A) Mouse spleens, mesenteric LN, and livers from WT and ICOSL KO mice infected with or without S. japonicum were harvested, and cells were stained with CD3-percpcy5.5, CD4-FITC, CXCR5-APC, and PD-1-PE antibodies. Data shown are gated on CD3+CD4+ cells. Numbers represent the frequency of the boxed population within the CD4+ T cell population; (B) Data are expressed as the mean ± SD of 18 mice from three independent experiments, ***, P<0.001 (Student's t-test); (C) The absolute numbers of CXCR5highPD-1high cells in spleens, mesenteric LN, and livers from WT and ICOSL KO mice infected with or without S. japonicum were calculated. Data are expressed as the mean ± SD of 18 mice from three independent experiments, ***, P<0.001 (Student's t-test); (D) Three weeks after adoptive transfer of none, PBS, non-Tfh CD4+ T cells, Tfh cells, or Th2 as described in Materials and Methods, liver sections from S. japonicum-infected ICOSL KO recipient mice were Paraffin-embedded, formalin fixed and stained with H&E. Images shown are representative of two independent experiments. Original magnification, ×100; (E) For each mouse, the sizes of 30 granulomas around single eggs were quantified with AxioVision Rel 4.7. Data are expressed in area units. Values are given as mean ± SD of 12 mice from two independent experiments, ***, P<0.001 (Student's t-test), compared with control mice; (F) Serum samples were collected from mice three weeks after adoptive transfer of none, PBS, Tfh cells, non-Tfh CD4+ T cells, or Th2 cells. Levels of serum ALT/AST were determined. Data are expressed as the mean ± SD of 12 mice from two independent experiments, **, P<0.01; *, P<0.05 (Student's t-test); (G) Paraffin-embedded sections were stained with sirius red. Images shown are representative of two independent experiments. Original magnification, ×100; (H) The mean optical density of collagen fibers by sirius red staining was digitized and analyzed on Image-Pro Plus software. Values are given as mean ± SD of 12 mice from two independent experiments, ***, P<0.001 (Student's t-test), compared with control mice.