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CD200R/CD200 inhibits osteoclastogenesis: new mechanism of osteoclast control by mesenchymal stem cells in human.

Varin A, Pontikoglou C, Labat E, Deschaseaux F, Sensebé L - PLoS ONE (2013)

Bottom Line: In this study, we demonstrated that soluble CD200 inhibited the differentiation of osteoclast precursors as well as their maturation in bone-resorbing cells in vitro.Soluble CD200 did not modify the monocyte phenotype but inhibited the receptor activator of nuclear factor kappa-B ligand (RANKL) signaling pathway as well as the gene expression of osteoclast markers such as osteoclast-associated receptor (OSCAR) and nuclear factor of activated T cells cytoplasmic 1 (NFATc1).Moreover, MSCs inhibited osteoclast formation, which depended on cell-cell contact and was associated with CD200 expression on the MSC surface.

View Article: PubMed Central - PubMed

Affiliation: STROMALab, UMR Univ. P. Sabatier/CNRS 5273, INSERM U1031, Toulouse, France. audrey.varin@efs.sante.fr

ABSTRACT
Bone homeostasis is maintained by the balance between bone-forming osteoblasts and bone-degrading osteoclasts. Osteoblasts have a mesenchymal origin whereas osteoclasts belong to the myeloid lineage. Osteoclast and osteoblast communication occurs through soluble factors secretion, cell-bone interaction and cell-cell contact, which modulate their activities. CD200 is an immunoglobulin superfamilly member expressed on various types of cells including mesenchymal stem cells (MSCs). CD200 receptor (CD200R) is expressed on myeloid cells such as monocytes/macrophages. We assume that CD200 could be a new molecule involved in the control of osteoclastogenesis and could play a role in MSC-osteoclast communication in humans. In this study, we demonstrated that soluble CD200 inhibited the differentiation of osteoclast precursors as well as their maturation in bone-resorbing cells in vitro. Soluble CD200 did not modify the monocyte phenotype but inhibited the receptor activator of nuclear factor kappa-B ligand (RANKL) signaling pathway as well as the gene expression of osteoclast markers such as osteoclast-associated receptor (OSCAR) and nuclear factor of activated T cells cytoplasmic 1 (NFATc1). Moreover, MSCs inhibited osteoclast formation, which depended on cell-cell contact and was associated with CD200 expression on the MSC surface. Our results clearly demonstrate that MSCs, through the expression of CD200, play a major role in the regulation of bone resorption and bone physiology and that the CD200-CD200R couple could be a new target to control bone diseases.

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Related in: MedlinePlus

Recombinant CD200 inhibits osteoclast formation.(A) Adherent peripheral blood mononuclear cells (PBMCs) were cultured with M-CSF for 48 hr, RANKL was added on day 3 of culture and medium was changed every 3 days. rCD200 was added at the beginning of the culture and every 3 days. After 21 days of culture, osteoclast formation was determined by the use of tartrate-resistant acid phosphatase (TRAP) assay. Data are mean±SD area of osteoclasts from 3 independent experiments. (B) rCD200 inhibits bone resorption activity. PBMCs were plated on bone slices in wells of 96-well plates and stimulated as described. After 21 days of culture, resorbtion pitt number and area were count under a light microscope (arrows). **, P<0.01 compared with none; *, P<0.05 compared with M+R alone.
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pone-0072831-g001: Recombinant CD200 inhibits osteoclast formation.(A) Adherent peripheral blood mononuclear cells (PBMCs) were cultured with M-CSF for 48 hr, RANKL was added on day 3 of culture and medium was changed every 3 days. rCD200 was added at the beginning of the culture and every 3 days. After 21 days of culture, osteoclast formation was determined by the use of tartrate-resistant acid phosphatase (TRAP) assay. Data are mean±SD area of osteoclasts from 3 independent experiments. (B) rCD200 inhibits bone resorption activity. PBMCs were plated on bone slices in wells of 96-well plates and stimulated as described. After 21 days of culture, resorbtion pitt number and area were count under a light microscope (arrows). **, P<0.01 compared with none; *, P<0.05 compared with M+R alone.

Mentions: The CD200–CD200R interaction can deliver an inhibitory signal to monocytes/macrophages [18,19]. To determine the effect of CD200 on osteoclastogenesis, we used a validated culture system: PBMCs were cultivated for 2 days with M-CSF to induce RANK expression and then treated with RANKL + M-CSF for 21 days to induce the formation of multinucleated TRAP+ osteoclast-like cells capable of resorbing bone surface (Figure 1A,B). The addition of different concentrations of rCD200 to the culture medium inhibited the RANKL-induced formation of osteoclasts (Figure 1A and Figure S1) and decreased the size and number of resorption pits on the bone surface (Figure 1B). Thus, rCD200 inhibited osteoclast formation and decreased their bone degradation capacity.


CD200R/CD200 inhibits osteoclastogenesis: new mechanism of osteoclast control by mesenchymal stem cells in human.

Varin A, Pontikoglou C, Labat E, Deschaseaux F, Sensebé L - PLoS ONE (2013)

Recombinant CD200 inhibits osteoclast formation.(A) Adherent peripheral blood mononuclear cells (PBMCs) were cultured with M-CSF for 48 hr, RANKL was added on day 3 of culture and medium was changed every 3 days. rCD200 was added at the beginning of the culture and every 3 days. After 21 days of culture, osteoclast formation was determined by the use of tartrate-resistant acid phosphatase (TRAP) assay. Data are mean±SD area of osteoclasts from 3 independent experiments. (B) rCD200 inhibits bone resorption activity. PBMCs were plated on bone slices in wells of 96-well plates and stimulated as described. After 21 days of culture, resorbtion pitt number and area were count under a light microscope (arrows). **, P<0.01 compared with none; *, P<0.05 compared with M+R alone.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3733817&req=5

pone-0072831-g001: Recombinant CD200 inhibits osteoclast formation.(A) Adherent peripheral blood mononuclear cells (PBMCs) were cultured with M-CSF for 48 hr, RANKL was added on day 3 of culture and medium was changed every 3 days. rCD200 was added at the beginning of the culture and every 3 days. After 21 days of culture, osteoclast formation was determined by the use of tartrate-resistant acid phosphatase (TRAP) assay. Data are mean±SD area of osteoclasts from 3 independent experiments. (B) rCD200 inhibits bone resorption activity. PBMCs were plated on bone slices in wells of 96-well plates and stimulated as described. After 21 days of culture, resorbtion pitt number and area were count under a light microscope (arrows). **, P<0.01 compared with none; *, P<0.05 compared with M+R alone.
Mentions: The CD200–CD200R interaction can deliver an inhibitory signal to monocytes/macrophages [18,19]. To determine the effect of CD200 on osteoclastogenesis, we used a validated culture system: PBMCs were cultivated for 2 days with M-CSF to induce RANK expression and then treated with RANKL + M-CSF for 21 days to induce the formation of multinucleated TRAP+ osteoclast-like cells capable of resorbing bone surface (Figure 1A,B). The addition of different concentrations of rCD200 to the culture medium inhibited the RANKL-induced formation of osteoclasts (Figure 1A and Figure S1) and decreased the size and number of resorption pits on the bone surface (Figure 1B). Thus, rCD200 inhibited osteoclast formation and decreased their bone degradation capacity.

Bottom Line: In this study, we demonstrated that soluble CD200 inhibited the differentiation of osteoclast precursors as well as their maturation in bone-resorbing cells in vitro.Soluble CD200 did not modify the monocyte phenotype but inhibited the receptor activator of nuclear factor kappa-B ligand (RANKL) signaling pathway as well as the gene expression of osteoclast markers such as osteoclast-associated receptor (OSCAR) and nuclear factor of activated T cells cytoplasmic 1 (NFATc1).Moreover, MSCs inhibited osteoclast formation, which depended on cell-cell contact and was associated with CD200 expression on the MSC surface.

View Article: PubMed Central - PubMed

Affiliation: STROMALab, UMR Univ. P. Sabatier/CNRS 5273, INSERM U1031, Toulouse, France. audrey.varin@efs.sante.fr

ABSTRACT
Bone homeostasis is maintained by the balance between bone-forming osteoblasts and bone-degrading osteoclasts. Osteoblasts have a mesenchymal origin whereas osteoclasts belong to the myeloid lineage. Osteoclast and osteoblast communication occurs through soluble factors secretion, cell-bone interaction and cell-cell contact, which modulate their activities. CD200 is an immunoglobulin superfamilly member expressed on various types of cells including mesenchymal stem cells (MSCs). CD200 receptor (CD200R) is expressed on myeloid cells such as monocytes/macrophages. We assume that CD200 could be a new molecule involved in the control of osteoclastogenesis and could play a role in MSC-osteoclast communication in humans. In this study, we demonstrated that soluble CD200 inhibited the differentiation of osteoclast precursors as well as their maturation in bone-resorbing cells in vitro. Soluble CD200 did not modify the monocyte phenotype but inhibited the receptor activator of nuclear factor kappa-B ligand (RANKL) signaling pathway as well as the gene expression of osteoclast markers such as osteoclast-associated receptor (OSCAR) and nuclear factor of activated T cells cytoplasmic 1 (NFATc1). Moreover, MSCs inhibited osteoclast formation, which depended on cell-cell contact and was associated with CD200 expression on the MSC surface. Our results clearly demonstrate that MSCs, through the expression of CD200, play a major role in the regulation of bone resorption and bone physiology and that the CD200-CD200R couple could be a new target to control bone diseases.

Show MeSH
Related in: MedlinePlus