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. 2016 Nov 29;11(11):e0167324.
doi: 10.1371/journal.pone.0167324. eCollection 2016.

Receptor-Type Protein-Tyrosine Phosphatase ζ and Colony Stimulating Factor-1 Receptor in the Intestine: Cellular Expression and Cytokine- And Chemokine Responses by Interleukin-34 and Colony Stimulating Factor-1

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Free PMC article

Receptor-Type Protein-Tyrosine Phosphatase ζ and Colony Stimulating Factor-1 Receptor in the Intestine: Cellular Expression and Cytokine- And Chemokine Responses by Interleukin-34 and Colony Stimulating Factor-1

Stephanie Zwicker et al. PLoS One. .
Free PMC article

Abstract

Differential intestinal expression of the macrophage growth factors colony stimulating factor-1 (CSF-1), interleukin (IL)-34, and their shared CSF-1 receptor (CSF-1R) in inflammatory bowel disease (IBD) has been shown. Diverse expression between CSF-1 and IL-34, suggest that IL-34 may signal via an alternate receptor. Receptor-type protein-tyrosine phosphatase ζ (PTPRZ1, RPTP-ζ), an additional IL-34 receptor, was recently identified. Here, we aimed to assess PTPRZ1 expression in IBD and non-IBD intestinal biopsies. Further, we aimed to investigate cellular PTPRZ1 and CSF-1R expression, and cytokine- and chemokine responses by IL-34 and CSF-1. The expression of PTPRZ1 was higher in non-IBD colon compared to ileum. PTPRZ1 expression was not altered with inflammation in IBD, however, correlated to IL34, CSF1, and CSF1R. The expression patterns of PTPRZ1 and CSF-1R differed in peripheral blood mononuclear cells (PBMCs), monocytes, macrophages, and intestinal epithelial cell line. PBMCs and monocytes of the same donors responded differently to IL-34 and CSF-1 with altered expression of tumor-necrosis factor α (TNF-α), IL-1β, interferon γ (IFN-γ), IL-13, IL-8, and monocyte chemotactic protein-1 (MCP-1) levels. This study shows that PTPRZ1 was expressed in bowel tissue. Furthermore, CSF-1R protein was detected in an intestinal epithelial cell line and donor dependently in primary PBMCs, monocytes, and macrophages, and first hints also suggest an expression in these cells for PTPRZ1, which may mediate IL-34 and CSF-1 actions.

Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. PTPRZ1 is differentially expressed in normal human ileum and colon but not regulated with inflammation.
(A) PTPRZ1 relative mRNA expression in ileum and colon, presented as the mean per colon sites for each non-IBD subjects. (B) PTPRZ1 relative mRNA in different sites of the colon from non-IBD subjects. PTPRZ1 relative mRNA expression in (C) colon and (D) ileum from non-IBD, IBD, UC and CD patients. Comparisons between ileum and colon were calculated using Mann–Whitney U tests, and between different sites of colon by ANOVA with post-hoc LSD tests. N = 18 for ileum, n = 24 for colon. Results are means ± SEM *P<0.05; **P<0.01; ***P<0.001. (E) PTPRZ1 relative mRNA expression compared between non-inflamed and inflamed in IBD patients. PTPRZ1 relative mRNA expression in colon presented as the mean per colon sites for each patient in IBD patients subdivided into CD and UC. Correlations of PTPRZ1 with (F) IL34, (G) CSF1, (H) CSF1R, (I) TNFA and (J) CD68 in IBD patients. Comparisons were evaluated using Mann–Whitney U tests. Correlations were assessed by Spearman’s correlation coefficients. N = 18 for non-inflamed IBD, n = 23 for inflamed IBD, n = 6 for non-inflamed CD, n = 6 for inflamed CD, n = 11 for non-inflamed UC, n = 16 for inflamed UC. Results are mean ± SEM *P<0.05; **P<0.01; ***P<0.001.
Fig 2
Fig 2. Expression of CSF-1R and PTPRZ1 in PBMCs, monocytes, macrophages and colonic epithelial cells.
Immunoblotting of lysates from PBMCs, monocytes, CSF-1 monocyte derived and non-polarized macrophages, Caco-2 and A549 cells analysed for (A) CSF-1R and (B) PTPRZ1. β-actin was used as a loading control (10 and 15 μg protein per lane, respectively).
Fig 3
Fig 3. Regulation of pro—inflammatory cytokines and chemokines through IL-34 and CSF-1 in PBMCs and monocytes IL1B, TNFA, IFNG, IL8 and MCP1 relative mRNA expression in PBMCs.
(A, B) and monocytes (C, D) stimulated with IL-34, CSF-1 for 1 h or left untreated were analysed by q-PCR and normalized to GAPDH. IL1B, TNFA, IFNG, IL8 and MCP1 relative mRNA expression from PBMCs (E, F) and monocytes (G, H) stimulated with IL-34, CSF-1 for 6 h or left untreated were analysed by q-PCR and normalized to GAPDH. Data represent mean + SEM, *P<0.05; **P<0.01; ***P<0.001, Student’s T-test, n = 5–6 donors. (I) Secreted MCP-1 in supernatants from Caco-2 cells, PBMCs and monocytes stimulated with IL-34 or CSF-1 for 24 h, as analysed by ELISA. Data represent mean + SEM, *P< 0.05; **P< 0.01; ***P< 0.001, Student’s T-test, n = 5–6 donors. (J) Secreted MCP-1 in supernatants from PBMCs stimulated with IL-34 or CSF-1 (10 or 50 ng/ml for 24h) were analysed by ELISA. Data represent mean + SEM, *P<0.05; **P<0.01; ***P<0.001, Student’s T-test, n = 6 donors.
Fig 4
Fig 4. CSF-1R dependent regulation of pro-inflammatory cytokines and chemokines in PBMCs and monocytes IL1B, TNFA, IFNG, IL8 and MCP1 relative mRNA expression from PBMCs.
(A, B) and monocytes (C, D) stimulated with IL-34, PBMCs (E, F) and monocytes (G, H) stimulated with CSF-1, after blocking CSF-1R for 6 h. IgG1 was used as a control. Gene expression was analysed by q-PCR and normalized to GAPDH. Data represent mean + SEM, *P<0.05; **P<0.01; ***P<0.001, Student’s T-test, n = 5–6 donors. (I-J) Secreted MCP-1 in supernatants from PBMCs (I) and monocytes (J) stimulated with IL-34 or CSF-1 after blocking CSF-1R for 24 h, IgG1 was used as a control, were analysed by ELISA. Data represent mean + SEM, *P<0.05; **P<0.01; ***P<0.001, Student’s T-test, n = 6 donors.
Fig 5
Fig 5. Diverse regulation of IL-10, IL-1β, TNF-α and MCP-1 in macrophages differentiated in the presence of IL-34 and CSF-1.
(A-C) IL1B, TNFA and IL10 mRNA expression in IL-34 and/or CSF-1 differentiated macrophages and polarized to an M1-like phenotype, an M2-like phenotype or non-polarized were analysed by q-PCR and normalized to GAPDH. Data represent mean + SEM, *P<0.05; **P<0.01; ***P<0.001, ANOVA, n = 5–6 donors. (D-F) Secreted IL-10, MCP-1 and IL-1β in the supernatant from IL-34 and/or CSF-1 differentiated macrophages and polarized to an M1-like phenotype, an M2-like phenotype or non-polarized were analysed by ELISA. Data represent mean + SEM, *P<0.05; **P< 0.01; ***P<0.001, ANOVA, n = 9 donors.

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Grant support

This work was supported by grants from the Swedish Research Council (523-2012-2480 to EAB), Stockholm County Council/ ALF project (20140367 to EAB), Karolinska Institutet Funds (EAB), Åke Wiberg Foundation (EAB) and the Ihre Foundation (SA). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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