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Follicular helper T cells promote liver pathology in mice during Schistosoma japonicum infection.

Chen X, Yang X, Li Y, Zhu J, Zhou S, Xu Z, He L, Xue X, Zhang W, Dong X, Wu H, Li CJ, Hsu HT, Kong W, Liu F, Tripathi PB, Yu MS, Chang J, Zhou L, Su C - PLoS Pathog. (2014)

Bottom Line: Following Schistosoma japonicum (S. japonicum) infection, granulomatous responses are induced by parasite eggs trapped in host organs, particular in the liver, during the acute stage of disease.While excessive liver granulomatous responses can lead to more severe fibrosis and circulatory impairment in chronically infected host.In addition, our results showed that the generation of Tfh cells driven by macrophages is dependent on cell-cell contact and the level of inducible costimulator ligand (ICOSL) on macrophages which is regulated by CD40-CD40L signaling.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathogen Biology & Immunology, Jiangsu Key Laboratory of Pathogen Biology, Nanjing Medical University, Nanjing, Jiangsu, P. R. China.

ABSTRACT
Following Schistosoma japonicum (S. japonicum) infection, granulomatous responses are induced by parasite eggs trapped in host organs, particular in the liver, during the acute stage of disease. While excessive liver granulomatous responses can lead to more severe fibrosis and circulatory impairment in chronically infected host. However, the exact mechanism of hepatic granuloma formation has remained obscure. In this study, we for the first time showed that follicular helper T (Tfh) cells are recruited to the liver to upregulate hepatic granuloma formation and liver injury in S. japonicum-infected mice, and identified a novel function of macrophages in Tfh cell induction. In addition, our results showed that the generation of Tfh cells driven by macrophages is dependent on cell-cell contact and the level of inducible costimulator ligand (ICOSL) on macrophages which is regulated by CD40-CD40L signaling. Our findings uncovered a previously unappreciated role for Tfh cells in liver pathology caused by S. japonicum infection in mice.

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Macrophages drive Tfh cell development through a cell-cell contact-dependent mechanism.(A, B) In a transwell system, macrophages from infected mice were cultured in triplicate wells in the lower chambers. CD4+ T cells from normal mice were cultured in the upper chambers. Expression of CXCR5 versus PD-1 on CD4+ T cells (gated as CD3+CD4+) after co-culture. Data are representative of two independent experiments. Numbers represent the frequency of the boxed population within the CD4+ T cell population. ***, P<0.001 (Student's t-test); (C) Flow cytometry of CD4+CXCR5+F4/80+ doublets in the spleens of S. japonicum-infected mice. Data are representative of three experiments with three mice in each group; (D) Gating strategy defining CD4+CXCR5+F4/80− singlet cells (R3), CD4−CXCR5−F4/80+ singlet cells (R2) or CD4+CXCR5+F4/80+ doublets (R4), (D right). Size (forward scatter (FSC)) of CD4+CXCR5+F4/80− singlet cells, CD4−CXCR5−F4/80+ singlet cells or CD4+CXCR5+F4/80+ doublets in the spleens of S. japonicum-infected mice (D left). Data are representative of three experiments. (E) Flow cytometry of CD3+CD4+CXCR5+F4/80+ doublets sorted from the pooled spleens from three S. japonicum-infected mice and then treated with 2 mM EDTA. Data are representative of three independent experiments.
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ppat-1004097-g005: Macrophages drive Tfh cell development through a cell-cell contact-dependent mechanism.(A, B) In a transwell system, macrophages from infected mice were cultured in triplicate wells in the lower chambers. CD4+ T cells from normal mice were cultured in the upper chambers. Expression of CXCR5 versus PD-1 on CD4+ T cells (gated as CD3+CD4+) after co-culture. Data are representative of two independent experiments. Numbers represent the frequency of the boxed population within the CD4+ T cell population. ***, P<0.001 (Student's t-test); (C) Flow cytometry of CD4+CXCR5+F4/80+ doublets in the spleens of S. japonicum-infected mice. Data are representative of three experiments with three mice in each group; (D) Gating strategy defining CD4+CXCR5+F4/80− singlet cells (R3), CD4−CXCR5−F4/80+ singlet cells (R2) or CD4+CXCR5+F4/80+ doublets (R4), (D right). Size (forward scatter (FSC)) of CD4+CXCR5+F4/80− singlet cells, CD4−CXCR5−F4/80+ singlet cells or CD4+CXCR5+F4/80+ doublets in the spleens of S. japonicum-infected mice (D left). Data are representative of three experiments. (E) Flow cytometry of CD3+CD4+CXCR5+F4/80+ doublets sorted from the pooled spleens from three S. japonicum-infected mice and then treated with 2 mM EDTA. Data are representative of three independent experiments.

Mentions: In parallel cultures, CD4+ T cells were separated from macrophages by a porous (0.4-µm) membrane in otherwise identical conditions. Results showed that compared to the co-culture group, the separation of CD4+ T cells from macrophages increased only the expression of PD-1, instead of CXCR5 (Figures 5A and 5B). Moreover, a small fraction of CXCR5+CD4+ T cells sorted from S. japonicum-infected mice spleens also expressed the macrophage marker F4/80, which suggested that these may represent stable macrophage-T cell conjugates (Figure 5C). In addition, the mean FSC value of the cells which expressed both T cell and macrophage markers was approximately 500, compared to T cells (CD4+CXCR5+F4/80−) and macrophages (CD4−CXCR5−F4/80+) that were approximately 200 and 300, respectively (Figure 5D), which suggested again that they were macrophage-T cell conjugates. Data showed that almost all CXCR5+CD4+F4/80+ cells expressed the T cell antigen receptor marker CD3, and further suggested that they were macrophage-T cell conjugates (Figure 5D). Next, we sorted the CD3+CXCR5+CD4+F4/80+ cells to high purity and then treated the cells with EDTA to dissociate cell-cell contacts. The resulting single cells segregated into approximately equal numbers of CD4+ and F4/80+ single-positive populations (Figure 5E), which further confirmed the stable conjugates. The macrophage-T cell conjugates in S. japonicum-infected mice livers were also detected (Figure S5). Taken together, these data suggest that direct cell-cell contact is required for Tfh cell development driven by macrophages, and also suggest a physiological role of these conjugates in vivo.


Follicular helper T cells promote liver pathology in mice during Schistosoma japonicum infection.

Chen X, Yang X, Li Y, Zhu J, Zhou S, Xu Z, He L, Xue X, Zhang W, Dong X, Wu H, Li CJ, Hsu HT, Kong W, Liu F, Tripathi PB, Yu MS, Chang J, Zhou L, Su C - PLoS Pathog. (2014)

Macrophages drive Tfh cell development through a cell-cell contact-dependent mechanism.(A, B) In a transwell system, macrophages from infected mice were cultured in triplicate wells in the lower chambers. CD4+ T cells from normal mice were cultured in the upper chambers. Expression of CXCR5 versus PD-1 on CD4+ T cells (gated as CD3+CD4+) after co-culture. Data are representative of two independent experiments. Numbers represent the frequency of the boxed population within the CD4+ T cell population. ***, P<0.001 (Student's t-test); (C) Flow cytometry of CD4+CXCR5+F4/80+ doublets in the spleens of S. japonicum-infected mice. Data are representative of three experiments with three mice in each group; (D) Gating strategy defining CD4+CXCR5+F4/80− singlet cells (R3), CD4−CXCR5−F4/80+ singlet cells (R2) or CD4+CXCR5+F4/80+ doublets (R4), (D right). Size (forward scatter (FSC)) of CD4+CXCR5+F4/80− singlet cells, CD4−CXCR5−F4/80+ singlet cells or CD4+CXCR5+F4/80+ doublets in the spleens of S. japonicum-infected mice (D left). Data are representative of three experiments. (E) Flow cytometry of CD3+CD4+CXCR5+F4/80+ doublets sorted from the pooled spleens from three S. japonicum-infected mice and then treated with 2 mM EDTA. Data are representative of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4006917&req=5

ppat-1004097-g005: Macrophages drive Tfh cell development through a cell-cell contact-dependent mechanism.(A, B) In a transwell system, macrophages from infected mice were cultured in triplicate wells in the lower chambers. CD4+ T cells from normal mice were cultured in the upper chambers. Expression of CXCR5 versus PD-1 on CD4+ T cells (gated as CD3+CD4+) after co-culture. Data are representative of two independent experiments. Numbers represent the frequency of the boxed population within the CD4+ T cell population. ***, P<0.001 (Student's t-test); (C) Flow cytometry of CD4+CXCR5+F4/80+ doublets in the spleens of S. japonicum-infected mice. Data are representative of three experiments with three mice in each group; (D) Gating strategy defining CD4+CXCR5+F4/80− singlet cells (R3), CD4−CXCR5−F4/80+ singlet cells (R2) or CD4+CXCR5+F4/80+ doublets (R4), (D right). Size (forward scatter (FSC)) of CD4+CXCR5+F4/80− singlet cells, CD4−CXCR5−F4/80+ singlet cells or CD4+CXCR5+F4/80+ doublets in the spleens of S. japonicum-infected mice (D left). Data are representative of three experiments. (E) Flow cytometry of CD3+CD4+CXCR5+F4/80+ doublets sorted from the pooled spleens from three S. japonicum-infected mice and then treated with 2 mM EDTA. Data are representative of three independent experiments.
Mentions: In parallel cultures, CD4+ T cells were separated from macrophages by a porous (0.4-µm) membrane in otherwise identical conditions. Results showed that compared to the co-culture group, the separation of CD4+ T cells from macrophages increased only the expression of PD-1, instead of CXCR5 (Figures 5A and 5B). Moreover, a small fraction of CXCR5+CD4+ T cells sorted from S. japonicum-infected mice spleens also expressed the macrophage marker F4/80, which suggested that these may represent stable macrophage-T cell conjugates (Figure 5C). In addition, the mean FSC value of the cells which expressed both T cell and macrophage markers was approximately 500, compared to T cells (CD4+CXCR5+F4/80−) and macrophages (CD4−CXCR5−F4/80+) that were approximately 200 and 300, respectively (Figure 5D), which suggested again that they were macrophage-T cell conjugates. Data showed that almost all CXCR5+CD4+F4/80+ cells expressed the T cell antigen receptor marker CD3, and further suggested that they were macrophage-T cell conjugates (Figure 5D). Next, we sorted the CD3+CXCR5+CD4+F4/80+ cells to high purity and then treated the cells with EDTA to dissociate cell-cell contacts. The resulting single cells segregated into approximately equal numbers of CD4+ and F4/80+ single-positive populations (Figure 5E), which further confirmed the stable conjugates. The macrophage-T cell conjugates in S. japonicum-infected mice livers were also detected (Figure S5). Taken together, these data suggest that direct cell-cell contact is required for Tfh cell development driven by macrophages, and also suggest a physiological role of these conjugates in vivo.

Bottom Line: Following Schistosoma japonicum (S. japonicum) infection, granulomatous responses are induced by parasite eggs trapped in host organs, particular in the liver, during the acute stage of disease.While excessive liver granulomatous responses can lead to more severe fibrosis and circulatory impairment in chronically infected host.In addition, our results showed that the generation of Tfh cells driven by macrophages is dependent on cell-cell contact and the level of inducible costimulator ligand (ICOSL) on macrophages which is regulated by CD40-CD40L signaling.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathogen Biology & Immunology, Jiangsu Key Laboratory of Pathogen Biology, Nanjing Medical University, Nanjing, Jiangsu, P. R. China.

ABSTRACT
Following Schistosoma japonicum (S. japonicum) infection, granulomatous responses are induced by parasite eggs trapped in host organs, particular in the liver, during the acute stage of disease. While excessive liver granulomatous responses can lead to more severe fibrosis and circulatory impairment in chronically infected host. However, the exact mechanism of hepatic granuloma formation has remained obscure. In this study, we for the first time showed that follicular helper T (Tfh) cells are recruited to the liver to upregulate hepatic granuloma formation and liver injury in S. japonicum-infected mice, and identified a novel function of macrophages in Tfh cell induction. In addition, our results showed that the generation of Tfh cells driven by macrophages is dependent on cell-cell contact and the level of inducible costimulator ligand (ICOSL) on macrophages which is regulated by CD40-CD40L signaling. Our findings uncovered a previously unappreciated role for Tfh cells in liver pathology caused by S. japonicum infection in mice.

Show MeSH
Related in: MedlinePlus