R Wiegertjes1, A van Caam1, H van Beuningen1, M Koenders1, P van Lent1, P van der Kraan1, F van de Loo1, E Blaney Davidson2. 1. Department of Experimental Rheumatology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands. 2. Department of Experimental Rheumatology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands. Electronic address: Esmeralda.BlaneyDavidson@radboudumc.nl.
Abstract
OBJECTIVE: Transforming growth factor-β (TGF-β) is an important homeostatic regulator of cartilage. In contrast, interleukin-6 (IL-6) is a pro-inflammatory cytokine implicated in cartilage degeneration. Cross-talk between TGF-β and IL-6 is reported in tissues other than articular cartilage. Here, we investigated regulation of IL-6 signaling by TGF-β in articular chondrocytes. DESIGN: Human primary chondrocytes and the human G6 chondrocyte cell line were stimulated with TGF-β1 or interleukin-1β (IL-1β). Expression of IL-6 and IL-6 receptor (IL-6R) was determined on mRNA and protein level. TGF-β regulation of IL-6 signaling via phosho-STAT3 (p-STAT3) was determined using Western blot, in presence of inhibitors for IL-6R, and Janus kinase(JAK)- and activin receptor-like kinase ALK)5 kinase activity. Furthermore, induction of STAT3-responsive genes was used as a read-out for IL-6 induced gene expression. RESULTS: TGF-β1 increased IL-6 mRNA and protein expression in both G6 and primary chondrocytes. Moreover, TGF-β1 stimulation clearly induced p-STAT3), which was abolished by inhibition of either IL-6R, JAK- or ALK5 kinase activity. However, TGF-β1 did not increase expression of the STAT3-responsive gene SOCS3 and pre-treatment with TGF-β1 even inhibited induction of p-STAT3 and SOCS3 by rhIL-6. Interestingly, TGF-β1 potently decreased IL-6R expression. In contrast, IL-1β did increase IL-6 levels, but did not affect IL-6R expression. Finally, addition of recombinant IL-6R abolished the inhibitory effect of TGF-β1 on IL-6-induced p-STAT3 and downstream SOCS3, BCL3, SAA1 and MMP1 expression. CONCLUSIONS: In this study we show that TGF-β decreases IL-6R expression, thereby dampening IL-6 signaling in chondrocytes. This reveals a novel effect of TGF-β, possibly important to restrict pro-inflammatory IL-6 effects to preserve cartilage homeostasis.
OBJECTIVE: Transforming growth factor-β (TGF-β) is an important homeostatic regulator of cartilage. In contrast, interleukin-6 (IL-6) is a pro-inflammatory cytokine implicated in cartilage degeneration. Cross-talk between TGF-β and IL-6 is reported in tissues other than articular cartilage. Here, we investigated regulation of IL-6 signaling by TGF-β in articular chondrocytes. DESIGN:Human primary chondrocytes and the human G6 chondrocyte cell line were stimulated with TGF-β1 or interleukin-1β (IL-1β). Expression of IL-6 and IL-6 receptor (IL-6R) was determined on mRNA and protein level. TGF-β regulation of IL-6 signaling via phosho-STAT3 (p-STAT3) was determined using Western blot, in presence of inhibitors for IL-6R, and Janus kinase(JAK)- and activin receptor-like kinase ALK)5 kinase activity. Furthermore, induction of STAT3-responsive genes was used as a read-out for IL-6 induced gene expression. RESULTS: TGF-β1 increased IL-6 mRNA and protein expression in both G6 and primary chondrocytes. Moreover, TGF-β1 stimulation clearly induced p-STAT3), which was abolished by inhibition of either IL-6R, JAK- or ALK5 kinase activity. However, TGF-β1 did not increase expression of the STAT3-responsive gene SOCS3 and pre-treatment with TGF-β1 even inhibited induction of p-STAT3 and SOCS3 by rhIL-6. Interestingly, TGF-β1 potently decreased IL-6R expression. In contrast, IL-1β did increase IL-6 levels, but did not affect IL-6R expression. Finally, addition of recombinant IL-6R abolished the inhibitory effect of TGF-β1 on IL-6-induced p-STAT3 and downstream SOCS3, BCL3, SAA1 and MMP1 expression. CONCLUSIONS: In this study we show that TGF-β decreases IL-6R expression, thereby dampening IL-6 signaling in chondrocytes. This reveals a novel effect of TGF-β, possibly important to restrict pro-inflammatory IL-6 effects to preserve cartilage homeostasis.
Authors: Eliana Lara-Barba; María Jesús Araya; Charlotte Nicole Hill; Felipe A Bustamante-Barrientos; Alexander Ortloff; Cynthia García; Felipe Galvez-Jiron; Carolina Pradenas; Noymar Luque-Campos; Gabriela Maita; Roberto Elizondo-Vega; Farida Djouad; Ana María Vega-Letter; Patricia Luz-Crawford Journal: Front Immunol Date: 2021-11-01 Impact factor: 7.561
Authors: Clara Sanjurjo-Rodriguez; Ala Altaie; Simon Mastbergen; Thomas Baboolal; Tim Welting; Floris Lafeber; Hemant Pandit; Dennis McGonagle; Elena Jones Journal: Front Bioeng Biotechnol Date: 2020-10-14