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Interleukin-17A upregulates receptor activator of NF-kappaB on osteoclast precursors.

Adamopoulos IE, Chao CC, Geissler R, Laface D, Blumenschein W, Iwakura Y, McClanahan T, Bowman EP - Arthritis Res. Ther. (2010)

Bottom Line: In this paper we describe a new mechanism by which the immune cytokine IL-17A directly affects osteoclastogenesis.Human CD14+ cells were isolated from healthy donors, cultured on dentine slices and coverslips and stimulated with IL-17A and/or receptor activator of NF-kappaB ligand (RANKL).IL-17A upregulates the receptor activator for NF-kappaB receptor on human osteoclast precursors in vitro, leading to increased sensitivity to RANKL signalling, osteoclast differentiation and bone loss.

View Article: PubMed Central - HTML - PubMed

Affiliation: Discovery Research, Schering-Plough Biopharma (formerly DNAX Research, Inc,), 901 South California Avenue, Palo Alto, CA 94304, USA. iannis.adamopoulos@spcorp.com

ABSTRACT

Introduction: The interaction between the immune and skeletal systems is evidenced by the bone loss observed in autoimmune diseases such as rheumatoid arthritis. In this paper we describe a new mechanism by which the immune cytokine IL-17A directly affects osteoclastogenesis.

Methods: Human CD14+ cells were isolated from healthy donors, cultured on dentine slices and coverslips and stimulated with IL-17A and/or receptor activator of NF-kappaB ligand (RANKL). Osteoclast differentiation was evaluated by gene expression, flow cytometry, tartrate-resistant acid phosphatase staining, fluorescence and electron microscopy. Physiologic bone remodelling was studied in wild-type (Wt) and IL-17A-/- mice using micro-computer tomography and serum RANKL/osteoprotegerin concentration. Functional osteoclastogenesis assays were performed using bone marrow macrophages isolated from IL-17A-/- and Wt mice.

Results: IL-17A upregulates the receptor activator for NF-kappaB receptor on human osteoclast precursors in vitro, leading to increased sensitivity to RANKL signalling, osteoclast differentiation and bone loss. IL-17A-/- mice have physiological bone homeostasis indistinguishable from Wt mice, and bone marrow macrophages isolated from these mice develop fully functional normal osteoclasts.

Conclusions: Collectively our data demonstrate anti-IL-17A treatment as a selective therapeutic target for bone loss associated with autoimmune diseases.

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IL17A-/- mice have normal bone mineral densities and osteoclast formation. (a) High-resolution micro-computer tomography analysis of 8-week-old male mouse femur midshaft and distal trabecular bone from IL-17A and wild-type (Wt) mice. (b) Serum receptor activator of NF-κB ligand (RANKL)/osteoprotegerin (OPG) levels of 8-week-old to 10-week-old IL-17A-/- mice and control mice. (c) Upper: bone marrow macrophages isolated from IL-17A-/- mice cultured for 6 days in the presence of macrophage colony-stimulating factor (MCSF)and RANKL form multinucleated tartrate-resistant acid phosphatase (TRAP)+ cells. Bars: 50 μm. Lower: Phalloidin, DAPI staining, and merged image of both stains of bone marrow macrophages isolated from IL-17A-/- mice and control mice cultured for 6 days in the presence of MCSF and RANKL showing F-actin ring formation. Bars: 25 μm. (d) Scanning electron photomicrographs of BMM cultures showing mature osteoclast resorbing activity (resorbed dentine has a rough, lighter colour appearance). Bars: 50 μm. Representative data of three experiments performed in triplicate. (e) Mean percentage area of dentine resorption of IL-17A and Wt mice.
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Figure 4: IL17A-/- mice have normal bone mineral densities and osteoclast formation. (a) High-resolution micro-computer tomography analysis of 8-week-old male mouse femur midshaft and distal trabecular bone from IL-17A and wild-type (Wt) mice. (b) Serum receptor activator of NF-κB ligand (RANKL)/osteoprotegerin (OPG) levels of 8-week-old to 10-week-old IL-17A-/- mice and control mice. (c) Upper: bone marrow macrophages isolated from IL-17A-/- mice cultured for 6 days in the presence of macrophage colony-stimulating factor (MCSF)and RANKL form multinucleated tartrate-resistant acid phosphatase (TRAP)+ cells. Bars: 50 μm. Lower: Phalloidin, DAPI staining, and merged image of both stains of bone marrow macrophages isolated from IL-17A-/- mice and control mice cultured for 6 days in the presence of MCSF and RANKL showing F-actin ring formation. Bars: 25 μm. (d) Scanning electron photomicrographs of BMM cultures showing mature osteoclast resorbing activity (resorbed dentine has a rough, lighter colour appearance). Bars: 50 μm. Representative data of three experiments performed in triplicate. (e) Mean percentage area of dentine resorption of IL-17A and Wt mice.

Mentions: The bone mineral density of cortical and trabecular bone was not significantly different between Wt mice and IL-17A-/- mice (Figure 4a). Serum OPG and RANKL levels in IL-17A-/- mice were similar to Wt mice (Figure 4b). The bone marrow isolated from IL-17A-/- mice had equal numbers of CD11b+ cells (osteoclast precursors) (Figure 4c), and when macrophages were stimulated with M-CSF and RANKL they differentiated into multinucleated TRAP+ cells capable of F-actin ring formation and dentine resorption (Figure 4d, e) - thus fulfilling the functional characteristics of osteoclasts. No differences were observed in any stage of osteoclast differentiation, nor in the function or activity of mature cells, and quantitative gene expression analysis confirmed that all osteoclast-related genes were similarly upregulated in in vitro differentiation cultures from IL-17A-/- mice compared with Wt mice (data not shown).


Interleukin-17A upregulates receptor activator of NF-kappaB on osteoclast precursors.

Adamopoulos IE, Chao CC, Geissler R, Laface D, Blumenschein W, Iwakura Y, McClanahan T, Bowman EP - Arthritis Res. Ther. (2010)

IL17A-/- mice have normal bone mineral densities and osteoclast formation. (a) High-resolution micro-computer tomography analysis of 8-week-old male mouse femur midshaft and distal trabecular bone from IL-17A and wild-type (Wt) mice. (b) Serum receptor activator of NF-κB ligand (RANKL)/osteoprotegerin (OPG) levels of 8-week-old to 10-week-old IL-17A-/- mice and control mice. (c) Upper: bone marrow macrophages isolated from IL-17A-/- mice cultured for 6 days in the presence of macrophage colony-stimulating factor (MCSF)and RANKL form multinucleated tartrate-resistant acid phosphatase (TRAP)+ cells. Bars: 50 μm. Lower: Phalloidin, DAPI staining, and merged image of both stains of bone marrow macrophages isolated from IL-17A-/- mice and control mice cultured for 6 days in the presence of MCSF and RANKL showing F-actin ring formation. Bars: 25 μm. (d) Scanning electron photomicrographs of BMM cultures showing mature osteoclast resorbing activity (resorbed dentine has a rough, lighter colour appearance). Bars: 50 μm. Representative data of three experiments performed in triplicate. (e) Mean percentage area of dentine resorption of IL-17A and Wt mice.
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Figure 4: IL17A-/- mice have normal bone mineral densities and osteoclast formation. (a) High-resolution micro-computer tomography analysis of 8-week-old male mouse femur midshaft and distal trabecular bone from IL-17A and wild-type (Wt) mice. (b) Serum receptor activator of NF-κB ligand (RANKL)/osteoprotegerin (OPG) levels of 8-week-old to 10-week-old IL-17A-/- mice and control mice. (c) Upper: bone marrow macrophages isolated from IL-17A-/- mice cultured for 6 days in the presence of macrophage colony-stimulating factor (MCSF)and RANKL form multinucleated tartrate-resistant acid phosphatase (TRAP)+ cells. Bars: 50 μm. Lower: Phalloidin, DAPI staining, and merged image of both stains of bone marrow macrophages isolated from IL-17A-/- mice and control mice cultured for 6 days in the presence of MCSF and RANKL showing F-actin ring formation. Bars: 25 μm. (d) Scanning electron photomicrographs of BMM cultures showing mature osteoclast resorbing activity (resorbed dentine has a rough, lighter colour appearance). Bars: 50 μm. Representative data of three experiments performed in triplicate. (e) Mean percentage area of dentine resorption of IL-17A and Wt mice.
Mentions: The bone mineral density of cortical and trabecular bone was not significantly different between Wt mice and IL-17A-/- mice (Figure 4a). Serum OPG and RANKL levels in IL-17A-/- mice were similar to Wt mice (Figure 4b). The bone marrow isolated from IL-17A-/- mice had equal numbers of CD11b+ cells (osteoclast precursors) (Figure 4c), and when macrophages were stimulated with M-CSF and RANKL they differentiated into multinucleated TRAP+ cells capable of F-actin ring formation and dentine resorption (Figure 4d, e) - thus fulfilling the functional characteristics of osteoclasts. No differences were observed in any stage of osteoclast differentiation, nor in the function or activity of mature cells, and quantitative gene expression analysis confirmed that all osteoclast-related genes were similarly upregulated in in vitro differentiation cultures from IL-17A-/- mice compared with Wt mice (data not shown).

Bottom Line: In this paper we describe a new mechanism by which the immune cytokine IL-17A directly affects osteoclastogenesis.Human CD14+ cells were isolated from healthy donors, cultured on dentine slices and coverslips and stimulated with IL-17A and/or receptor activator of NF-kappaB ligand (RANKL).IL-17A upregulates the receptor activator for NF-kappaB receptor on human osteoclast precursors in vitro, leading to increased sensitivity to RANKL signalling, osteoclast differentiation and bone loss.

View Article: PubMed Central - HTML - PubMed

Affiliation: Discovery Research, Schering-Plough Biopharma (formerly DNAX Research, Inc,), 901 South California Avenue, Palo Alto, CA 94304, USA. iannis.adamopoulos@spcorp.com

ABSTRACT

Introduction: The interaction between the immune and skeletal systems is evidenced by the bone loss observed in autoimmune diseases such as rheumatoid arthritis. In this paper we describe a new mechanism by which the immune cytokine IL-17A directly affects osteoclastogenesis.

Methods: Human CD14+ cells were isolated from healthy donors, cultured on dentine slices and coverslips and stimulated with IL-17A and/or receptor activator of NF-kappaB ligand (RANKL). Osteoclast differentiation was evaluated by gene expression, flow cytometry, tartrate-resistant acid phosphatase staining, fluorescence and electron microscopy. Physiologic bone remodelling was studied in wild-type (Wt) and IL-17A-/- mice using micro-computer tomography and serum RANKL/osteoprotegerin concentration. Functional osteoclastogenesis assays were performed using bone marrow macrophages isolated from IL-17A-/- and Wt mice.

Results: IL-17A upregulates the receptor activator for NF-kappaB receptor on human osteoclast precursors in vitro, leading to increased sensitivity to RANKL signalling, osteoclast differentiation and bone loss. IL-17A-/- mice have physiological bone homeostasis indistinguishable from Wt mice, and bone marrow macrophages isolated from these mice develop fully functional normal osteoclasts.

Conclusions: Collectively our data demonstrate anti-IL-17A treatment as a selective therapeutic target for bone loss associated with autoimmune diseases.

Show MeSH
Related in: MedlinePlus