Protein Tyrosine Phosphatase Non-Receptor Type 2 Function in Dendritic Cells Is Crucial to Maintain Tissue Tolerance - PubMed (original) (raw)

doi: 10.3389/fimmu.2020.01856. eCollection 2020.

Egle Katkeviciute 1, Marlene Schwarzfischer 1, Philipp Busenhart 1, Claudia Gottier 1, Dunja Mrdjen 2, Juliana Komuczki 2, Marcin Wawrzyniak 1, Silvia Lang 1, Kirstin Atrott 1, Burkhard Becher 2, Gerhard Rogler 1 3, Michael Scharl 1 3, Marianne R Spalinger 1

Affiliations

Protein Tyrosine Phosphatase Non-Receptor Type 2 Function in Dendritic Cells Is Crucial to Maintain Tissue Tolerance

Larissa Hering et al. Front Immunol. 2020.

Abstract

Protein tyrosine phosphatase non-receptor type 2 (PTPN2) plays a pivotal role in immune homeostasis and has been associated with human autoimmune and chronic inflammatory diseases. Though PTPN2 is well-characterized in lymphocytes, little is known about its function in innate immune cells. Our findings demonstrate that dendritic cell (DC)-intrinsic PTPN2 might be the key to explain the central role for PTPN2 in the immune system to maintain immune tolerance. Partial genetic PTPN2 ablation in DCs resulted in spontaneous inflammation, particularly in skin, liver, lung and kidney 22 weeks post-birth. DC-specific PTPN2 controls steady-state immune cell composition and even incomplete PTPN2 deficiency in DCs resulted in enhanced organ infiltration of conventional type 2 DCs, accompanied by expansion of IFNγ-producing effector T-cells. Consequently, the phenotypic effects of DC-specific PTPN2 deficiency were abolished in T-cell deficient Rag knock-out mice. Our data add substantial knowledge about the molecular mechanisms to prevent inflammation and maintain tissue tolerance.

Keywords: IFNγ; PTPN2; dendritic cells; inflammatory diseases; loss of tolerance; systemic inflammation.

Copyright © 2020 Hering, Katkeviciute, Schwarzfischer, Busenhart, Gottier, Mrdjen, Komuczki, Wawrzyniak, Lang, Atrott, Becher, Rogler, Scharl and Spalinger.

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Figures

Figure 1

Figure 1

Altered immune cell populations upon partial PTPN2 deletion in DCs. Immune cells were analyzed in 5-weeks-old, 13-weeks-old, and 22-weeks-old PTPN2fl/fl (WT) and PTPN2fl/fl × CD11cCre (KO) mice. _t_-SNE maps displaying live, CD45+ single cells in skin, liver, lung, and kidney. Colors correspond to FlowSOM-guided clustering of cell populations. Pie charts represent relative numbers among CD45+ cells. (A) _t_-SNE maps of CD45+ cells in 5-weeks-old mice. (B) Total counts of CD45+ cells among indicated tissues in 5-weeks-old-mice. (C) Relative numbers of neutrophils, monocytes, macrophages, and DCs among CD45+ cells among indicated tissues in 5-weeks-old mice. (D,E) _t_-SNE maps of CD45+ cells in 13-weeks-old (D) and 22-weeks-old (E) mice. Data are representative of two independent experiments with n ≥ 4 mice (A–E). *p < 0.05; **p < 0.01 [two-tailed Mann Whitney test **(C)**]. Data are shown as mean ± s.d. (C).

Figure 2

Figure 2

Partial loss of PTPN2 increases cDC2 accumulation in skin and liver. (A) Representative flow cytometry plots for identification of CD11b+ cDC1s (brown) and CD24+ cDC2s (green). (B) IRF4 and IRF8 expression in cDC1s and cDC2s. (C) Pie charts of the proportion of cDC1s and cDC2s across tissues in young (5-weeks-old) PTPN2fl/fl (WT) and PTPN2fl/fl × CD11cCre (KO) mice. (D–F) Frequencies of cDC subset distribution in 5-weeks-old (D), 13-weeks-old (E), and 22-weeks-old (F) mice. Data are representative of two independent experiments with n ≥ 4 mice (C–F). *p < 0.05 [two-tailed Mann Whitney test **(D–F)**]. Data are shown as mean ± s.d. (D–F).

Figure 3

Figure 3

PTPN2 in DCs affects T cell infiltration and activation. (A) Absolute numbers of CD3+ T cells in PTPN2fl/fl (WT) and PTPN2fl/fl × CD11cCre (KO) mice. (B) Absolute numbers of CD4+ and CD8+ T cells. (C–F) tSNE map of CD3+ cells from 5-weeks-old mice in skin (C), liver (D), lung (E), and kidney (F). Colors correspond to FlowSOM-guided clustering of cell populations. (C–F) Relative frequencies of T cell subsets in skin (C), liver (D), lung (E), and kidney (F). (G) Manually gated IFNγ expression of CD4+ and CD8+ T cells. Data are representative of two independent experiments with n ≥ 4 mice (A–F). *p < 0.05; **p < 0.01 [two-tailed Mann Whitney test **(A–F)**]. Data are shown as mean ± s.d. (A–F).

Figure 4

Figure 4

Spontaneous inflammatory infiltrates in liver and skin upon partial ablation of PTPN2 in DCs. (A–C) Representative pictures from H&E-stained sections of skin (A), kidney (B), and liver (C) of 25-weeks-old PTPN2fl/fl (WT) and PTPN2fl/fl × CD11cCre (KO) mice. (D) Spleen weight. (E) Serum levels of IgG and anti-dsDNA IgG. (F) Serum levels of amino transferases. (G) Serum levels of inidcated cytokines. Each dot represents one mouse [n = 9 mice (D), n ≥ 3 mice (E), n ≥ 8 mice **(F,G)**]. *p < 0.05; ****p < 0.00001 [two-tailed Mann Whitney test **(D–G)**]. Data are shown as ± s.d. (D–G).

Figure 5

Figure 5

Tissue inflammation is dependent on lymphocytes. PTPN2fl/fl × CD11cCre mice were crossed to RAG2−/− mice to generate PTPN2fl/fl × CD11cCre × RAG−/− mice. (A) Frequencies of cDCs subsets in 5-weeks-old PTPN2fl/fl (WT), PTPN2fl/fl × CD11cCre (KO), PTPN2fl/fl × RAG−/− (WT RAG), and PTPN2fl/fl × CD11cCre × RAG−/− (KO RAG). (B–D) Representative pictures from H&E-stained sections of skin (B), kidney (C), and liver (D) in 25-weeks-old mice. (E) Spleen weight in 25-weeks-old mice. Data are representative of two independent experiments [n ≥ 4 mice (A), n ≥ 9 mice **(E)**]. *p < 0.05; **p < 0.01; ***p < 0.001 [two-tailed Mann Whitney test **(A,E)**]. Data are shown as mean ± s.d. (A,E).

Figure 6

Figure 6

Increased DC activation via upregulation of the IFNγ-STAT1 pathway. (A–C) BMDCs were generated from PTPN2fl/fl (WT) and PTPN2fl/fl × CD11cCre (KO) mice. (A,B) Phosphorylation levels of STAT1 and STAT3 determined by Western Blot (A) and Flow Cytometry (B). (C) mRNA expression of IFNγ pathway-associated genes. (D) mRNA expression levels of IRF1 in lysates of indicated organs. (E) Mean fluorescence intensity (MFI) of co-stimulatory molecules on cDCs. (F,G) Relative numbers of neutrophils and monocytes in skin (F) and liver (G) of 5-weeks-old mice after antibiotics treatment. Data are pooled from two independent experiments (A–C), representative of two independent experiments (D), pooled from three independent experiments (E), or pooled from three independent experiments (F,G) [n = 6 mice (A,B), n ≥ 3 mice (C), n ≥ 3 mice (D), n ≥ 10 mice (E), n ≥ 6 mice **(F,G)**]. *p < 0.05; **p < 0.01; ****p < 0.00001 [one-way ANOVA, Tukey's multiple comparison test (A–C,F,G); two-tailed Mann Whitney test **(D,E)**]. Data are shown as mean ± s.d (A–G).

References

    1. Iberg CA, Jones A, Hawiger D. Dendritic cells as inducers of peripheral tolerance. Trends Immunol. (2017) 38:793–804. 10.1016/j.it.2017.07.007 - DOI - PMC - PubMed
    1. Ganguly D, Haak S, Sisirak V, Reizis B. The role of dendritic cells in autoimmunity. Nat Rev Immunol. (2013) 13:566–77. 10.1038/nri3477 - DOI - PMC - PubMed
    1. Burton PR, Clayton DG, Cardon LR, Craddock N, Deloukas P, Duncanson A, et al. Genome-wide association study of 14,000 cases of seven common diseases and 3,000 shared controls. Nature. (2007) 447:661–78. 10.1038/nature05911 - DOI - PMC - PubMed
    1. Khor B, Gardet A, Xavier RJ. Genetics and pathogenesis of inflammatory bowel disease. Nature. (2011) 474:307–17. 10.1038/nature10209 - DOI - PMC - PubMed
    1. Todd JA, Walker NM, Cooper JD, Smyth DJ, Downes K, Plagnol V, et al. Robust associations of four new chromosome regions from genome-wide analyses of type 1 diabetes. Nat Genet. (2007) 39:857–64. 10.1038/ng2068 - DOI - PMC - PubMed

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