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Blockade of IL-36 receptor signaling does not prevent from TNF-induced arthritis.

Derer A, Groetsch B, Harre U, Böhm C, Towne J, Schett G, Frey S, Hueber AJ - PLoS ONE (2014)

Bottom Line: Furthermore, blockade of IL-36 signaling did not change histological signs of TNF-induced arthritis.Additionally, no alteration on bone homeostasis was observed in ex vivo murine and human osteoclast differentiation assays.Thus we conclude that IL-36α does not affect the development of inflammatory arthritis.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine 3 and Institute for Clinical Immunology, University Hospital Erlangen, Erlangen, Germany; Department of Radiation Oncology, University Hospital Erlangen, Erlangen, Germany.

ABSTRACT

Introduction: Interleukin (IL)-36α is a newly described member of the IL-1 cytokine family with a known inflammatory and pathogenic function in psoriasis. Recently, we could demonstrate that the receptor (IL-36R), its ligand IL-36α and its antagonist IL-36Ra are expressed in synovial tissue of arthritis patients. Furthermore, IL-36α induces MAP-kinase and NFκB signaling in human synovial fibroblasts with subsequent expression and secretion of pro-inflammatory cytokines.

Methods: To understand the pathomechanism of IL-36 dependent inflammation, we investigated the biological impact of IL-36α signaling in the hTNFtg mouse. Also the impact on osteoclastogenesis by IL-36α was tested in murine and human osteoclast assays.

Results: Diseased mice showed an increased expression of IL-36R and IL-36α in inflamed knee joints compared to wildtype controls. However, preventively treating mice with an IL-36R blocking antibody led to no changes in clinical onset and pattern of disease. Furthermore, blockade of IL-36 signaling did not change histological signs of TNF-induced arthritis. Additionally, no alteration on bone homeostasis was observed in ex vivo murine and human osteoclast differentiation assays.

Conclusion: Thus we conclude that IL-36α does not affect the development of inflammatory arthritis.

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Related in: MedlinePlus

Osteoclastogenesis is not influenced by IL-36α treatment.(A) Quantification of human osteoclast assays from four different donors, done in two independent experiments; each of them was done in triplicates, treated with rhIL36α alone, rhIL36α plus IL36RA or rhIL36α plus recombinant receptor. (B) Quantitative real-time PCR for CtsK, IL-1RAcP and IL-36R of human osteoclast precursor cells stimulated with M-CSF and RANKL to achieve osteoclast differentiation. Relative Expression was calculated from the ratio of the gene of interest to the housekeeping gene Hsp90ab1. Data from three (CtsK) or four (IL-1RAcP and IL-36R) independent donors are shown. (C) Representative pictures and quantification of three independent murine osteoclast assay untreated (control) or stimulated with 100 ng/ml recombinant murine IL36α. (D) Quantitative real-time PCR for CtsK and TRAP of murine osteoclast precursor cells stimulated with M-CSF, M-CSF and RANKL or M-CSF, RANKL and rmIL-36α to achieve osteoclast differentiation. Data from three independent experiments are shown. Relative expression was calculated from the ratio of the gene of interest to the housekeeping gene β-actin. Graphs depict mean ±SEM.
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pone-0101954-g005: Osteoclastogenesis is not influenced by IL-36α treatment.(A) Quantification of human osteoclast assays from four different donors, done in two independent experiments; each of them was done in triplicates, treated with rhIL36α alone, rhIL36α plus IL36RA or rhIL36α plus recombinant receptor. (B) Quantitative real-time PCR for CtsK, IL-1RAcP and IL-36R of human osteoclast precursor cells stimulated with M-CSF and RANKL to achieve osteoclast differentiation. Relative Expression was calculated from the ratio of the gene of interest to the housekeeping gene Hsp90ab1. Data from three (CtsK) or four (IL-1RAcP and IL-36R) independent donors are shown. (C) Representative pictures and quantification of three independent murine osteoclast assay untreated (control) or stimulated with 100 ng/ml recombinant murine IL36α. (D) Quantitative real-time PCR for CtsK and TRAP of murine osteoclast precursor cells stimulated with M-CSF, M-CSF and RANKL or M-CSF, RANKL and rmIL-36α to achieve osteoclast differentiation. Data from three independent experiments are shown. Relative expression was calculated from the ratio of the gene of interest to the housekeeping gene β-actin. Graphs depict mean ±SEM.

Mentions: To investigate the impact of IL-36 on bone homeostasis, we analyzed the effect of IL-36α on osteoclastogenesis. The stimulation of human osteoclast precursors with recombinant human IL-36α did not alter their differentiation into mature osteoclasts (Figure 5A). Additional blockade of IL-36α signaling via its antagonist IL-36Ra or the soluble IL-36 receptor did not have further impact (Figure 5A). Quantitative PCR analysis revealed a rapid down-regulation of the IL-1RAcP and the IL-36R during osteoclastogenesis (demonstrated by the up-regulation of Cathepsin K (CtsK) as an osteoclast-specific marker) (Figure 5B), suggesting the absence of the IL-36 receptor on mature osteoclasts. These data were confirmed in murine osteoclast assays with no influence of IL-36α on osteoclast differentiation (Figure 5C) or markers for osteoclastogenesis (Cathepsin K and TRAP, Figure 5D).


Blockade of IL-36 receptor signaling does not prevent from TNF-induced arthritis.

Derer A, Groetsch B, Harre U, Böhm C, Towne J, Schett G, Frey S, Hueber AJ - PLoS ONE (2014)

Osteoclastogenesis is not influenced by IL-36α treatment.(A) Quantification of human osteoclast assays from four different donors, done in two independent experiments; each of them was done in triplicates, treated with rhIL36α alone, rhIL36α plus IL36RA or rhIL36α plus recombinant receptor. (B) Quantitative real-time PCR for CtsK, IL-1RAcP and IL-36R of human osteoclast precursor cells stimulated with M-CSF and RANKL to achieve osteoclast differentiation. Relative Expression was calculated from the ratio of the gene of interest to the housekeeping gene Hsp90ab1. Data from three (CtsK) or four (IL-1RAcP and IL-36R) independent donors are shown. (C) Representative pictures and quantification of three independent murine osteoclast assay untreated (control) or stimulated with 100 ng/ml recombinant murine IL36α. (D) Quantitative real-time PCR for CtsK and TRAP of murine osteoclast precursor cells stimulated with M-CSF, M-CSF and RANKL or M-CSF, RANKL and rmIL-36α to achieve osteoclast differentiation. Data from three independent experiments are shown. Relative expression was calculated from the ratio of the gene of interest to the housekeeping gene β-actin. Graphs depict mean ±SEM.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4128584&req=5

pone-0101954-g005: Osteoclastogenesis is not influenced by IL-36α treatment.(A) Quantification of human osteoclast assays from four different donors, done in two independent experiments; each of them was done in triplicates, treated with rhIL36α alone, rhIL36α plus IL36RA or rhIL36α plus recombinant receptor. (B) Quantitative real-time PCR for CtsK, IL-1RAcP and IL-36R of human osteoclast precursor cells stimulated with M-CSF and RANKL to achieve osteoclast differentiation. Relative Expression was calculated from the ratio of the gene of interest to the housekeeping gene Hsp90ab1. Data from three (CtsK) or four (IL-1RAcP and IL-36R) independent donors are shown. (C) Representative pictures and quantification of three independent murine osteoclast assay untreated (control) or stimulated with 100 ng/ml recombinant murine IL36α. (D) Quantitative real-time PCR for CtsK and TRAP of murine osteoclast precursor cells stimulated with M-CSF, M-CSF and RANKL or M-CSF, RANKL and rmIL-36α to achieve osteoclast differentiation. Data from three independent experiments are shown. Relative expression was calculated from the ratio of the gene of interest to the housekeeping gene β-actin. Graphs depict mean ±SEM.
Mentions: To investigate the impact of IL-36 on bone homeostasis, we analyzed the effect of IL-36α on osteoclastogenesis. The stimulation of human osteoclast precursors with recombinant human IL-36α did not alter their differentiation into mature osteoclasts (Figure 5A). Additional blockade of IL-36α signaling via its antagonist IL-36Ra or the soluble IL-36 receptor did not have further impact (Figure 5A). Quantitative PCR analysis revealed a rapid down-regulation of the IL-1RAcP and the IL-36R during osteoclastogenesis (demonstrated by the up-regulation of Cathepsin K (CtsK) as an osteoclast-specific marker) (Figure 5B), suggesting the absence of the IL-36 receptor on mature osteoclasts. These data were confirmed in murine osteoclast assays with no influence of IL-36α on osteoclast differentiation (Figure 5C) or markers for osteoclastogenesis (Cathepsin K and TRAP, Figure 5D).

Bottom Line: Furthermore, blockade of IL-36 signaling did not change histological signs of TNF-induced arthritis.Additionally, no alteration on bone homeostasis was observed in ex vivo murine and human osteoclast differentiation assays.Thus we conclude that IL-36α does not affect the development of inflammatory arthritis.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine 3 and Institute for Clinical Immunology, University Hospital Erlangen, Erlangen, Germany; Department of Radiation Oncology, University Hospital Erlangen, Erlangen, Germany.

ABSTRACT

Introduction: Interleukin (IL)-36α is a newly described member of the IL-1 cytokine family with a known inflammatory and pathogenic function in psoriasis. Recently, we could demonstrate that the receptor (IL-36R), its ligand IL-36α and its antagonist IL-36Ra are expressed in synovial tissue of arthritis patients. Furthermore, IL-36α induces MAP-kinase and NFκB signaling in human synovial fibroblasts with subsequent expression and secretion of pro-inflammatory cytokines.

Methods: To understand the pathomechanism of IL-36 dependent inflammation, we investigated the biological impact of IL-36α signaling in the hTNFtg mouse. Also the impact on osteoclastogenesis by IL-36α was tested in murine and human osteoclast assays.

Results: Diseased mice showed an increased expression of IL-36R and IL-36α in inflamed knee joints compared to wildtype controls. However, preventively treating mice with an IL-36R blocking antibody led to no changes in clinical onset and pattern of disease. Furthermore, blockade of IL-36 signaling did not change histological signs of TNF-induced arthritis. Additionally, no alteration on bone homeostasis was observed in ex vivo murine and human osteoclast differentiation assays.

Conclusion: Thus we conclude that IL-36α does not affect the development of inflammatory arthritis.

Show MeSH
Related in: MedlinePlus