IL-6, leukemia inhibitory factor, and oncostatin M stimulate bone resorption and regulate the expression of receptor activator of NF-kappa B ligand, osteoprotegerin, and receptor activator of NF-kappa B in mouse calvariae.

Py Palmqvist,Ulf H Lerner,Emma Persson,H Herschel Conaway

doi:10.4049/jimmunol.169.6.3353

Py Palmqvist, Ulf H Lerner + Show 2 more

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https://doi.org/10.4049/jimmunol.169.6.3353

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Abstract

IL-6, leukemia inhibitory factor (LIF), and oncostatin M (OSM) are IL-6-type cytokines that stimulate osteoclast formation and function. In the present study, the resorptive effects of these agents and their regulation of receptor activator of NF-kappaB ligand (RANKL), RANK, and osteoprotegerin (OPG) were studied in neonatal mouse calvaria. When tested separately, neither human (h) IL-6 nor the human soluble IL-6R (shIL-6R) stimulated bone resorption, but when hIL-6 and the shIL-6R were combined, significant stimulation of both mineral and matrix release from bone explants was noted. Semiquantitative RT-PCR showed that hIL-6 plus shIL-6R enhanced the expression of RANKL and OPG in calvarial bones, but decreased RANK expression. Human LIF, hOSM, and mouse OSM (mOSM) also stimulated 45Ca release and enhanced the mRNA expression of RANKL and OPG in mouse calvaria, but had no effect on the expression of RANK. In agreement with the RT-PCR analyses, ELISA measurements showed that both hIL-6 plus shIL-6R and mOSM increased RANKL and OPG proteins. 1,25-Dihydroxyvitamin D3 (D3) also increased the RANKL protein level, but decreased the protein level of OPG. OPG inhibited 45Ca release stimulated by RANKL, hIL-6 plus shIL-6R, hLIF, hOSM, mOSM, and D3. An Ab neutralizing mouse gp130 inhibited 45Ca release induced by hIL-6 plus shIL-6R. These experiments demonstrated stimulation of calvarial bone resorption and regulation of mRNA and protein expression of RANKL and OPG by D3 and IL-6 family cytokines as well as regulation of RANK expression in preosteoclasts/osteoclasts of mouse calvaria by D3 and hIL-6 plus shIL-6R.

Highlights

ObjectivesThe aims of the present study were to: 1) determine whether IL-6 together with sIL-6R could stimulate bone resorption in neonatal mouse calvaria, 2) determine whether oncostatin M (OSM) had a resorptive action similar to that of leukemia inhibitory factor (LIF) in calvarial bones, and 3) assess the roles of RANKL, RANK, and OPG in the calvarial effects of IL-6, LIF, and OSM
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Recombinant human IL-6, recombinant mouse IL-6, recombinant human sIL-6R, recombinant human leukemia inhibitory factor (LIF), recombinant human oncostatin M (OSM), recombinant mouse OSM, recombinant human IL-1␤, recombinant mRANKL and mOPG fusion proteins, neutralizing anti-mouse gp130 Ab, and mOPG and mRANKL immunoassay kits were purchased from R&D Systems (Abingdon, U.K.); FBS was obtained from ICN Pharmaceuticals (Costa Mesa, CA); acetazolamide, hydroxyurea, and essentially acid free fetal BSA were purchased from Sigma-Aldrich

Summary

Objectives

The aims of the present study were to: 1) determine whether IL-6 together with sIL-6R could stimulate bone resorption in neonatal mouse calvaria, 2) determine whether OSM had a resorptive action similar to that of LIF in calvarial bones, and 3) assess the roles of RANKL, RANK, and OPG in the calvarial effects of IL-6, LIF, and OSM

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Results

Conclusion