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CD16 (FcRgammaIII) as a potential marker of osteoclast precursors in psoriatic arthritis.

Chiu YG, Shao T, Feng C, Mensah KA, Thullen M, Schwarz EM, Ritchlin CT - Arthritis Res. Ther. (2010)

Bottom Line: Exposure of cells to OC-promoting, but not DC-promoting media, was associated with CD16 up-regulation.An increased frequency of circulating CD14+CD16+ cells was noted in PsA compared to controls, and intermediate levels of CD16 may suggest a transitional state of OCP during osteoclastogenesis.Collectively, our data suggest that CD16 has the potential to serve as an OCP marker in inflammatory arthritis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Allergy/Immunology & Rheumatology Unit, University of Rochester Medical School, 601 Elmwood Avenue, Rochester, NY 14642, USA. grace_chiu@urmc.rochester.edu

ABSTRACT

Introduction: Psoriatic arthritis (PsA) is a chronic inflammatory arthritis characterized by bone erosion mediated by osteoclasts (OC). Our previous studies showed an elevated frequency of OC precursors (OCP) in PsA patients. Here, we examined if OC arise from CD16-positive monocytes in PsA.

Methods: Peripheral blood mononuclear cells (PBMC) or monocytes were isolated from human peripheral blood and sorted based on CD16 expression. Sorted cells were cultured alone or with bone wafers in the presence of receptor activator of nuclear factor kappa-B ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). Enumeration and bone erosion activity of OC were examined after culture. The effects of tumor necrosis factor-alpha (TNFalpha), OC-promoting (M-CSF plus RANKL), and dendritic cell (DC)-promoting (GM-CSF plus interleukin (IL)-4) cytokines on CD16 surface expression were examined by flow cytometry.

Results: PsA and psoriasis (Ps) subjects had a higher percentage of circulating inflammatory CD14+CD16+ cells than healthy controls (HC). Exposure of cells to OC-promoting, but not DC-promoting media, was associated with CD16 up-regulation. PBMC of Ps and PsA had a higher frequency of cells expressing intermediate levels of CD16. OC were mainly derived from CD16+ cells in PsA. Increased CD16 expression was associated with a higher bone erosion activity in PsA.

Conclusions: An increased frequency of circulating CD14+CD16+ cells was noted in PsA compared to controls, and intermediate levels of CD16 may suggest a transitional state of OCP during osteoclastogenesis. Intriguingly, TNFalpha blocked CD16 expression on a subset of CD14+ monocytes. Collectively, our data suggest that CD16 has the potential to serve as an OCP marker in inflammatory arthritis.

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Cytokines alter the cell surface expression of CD16 and human CD14+ cells undergo a transitional stage of CD16 up-regulation in OC-promoting culture conditions. (a) Enriched human monocytes were cultured in osteoclast (OC)-promoting media (receptor activator of nuclear factor kappa-B ligand (RANKL) + macrophage colony-stimulating factor (M-CSF), pink line) or dendritic cell (DC)-promoting media (IL-4 + granulocyte-macrophage colony-stimulating factor (GM-CSF), green line). Freshly isolated monocytes (blue line) and the isotype control (black line) are also shown. Surface expression of CD16 was monitored by FACS analysis on [a] day 0, [b] day 1, and [c] day 3, respectively. (b) Human peripheral blood mononuclear cells (PBMC) were cultured in OC-promoting media (RANKL + M-CSF) and the cell surface expression of CD14 and CD16 was monitored on [a] day 0, [b] day 3, and [c] day 5. Data shown here are live cells after forward scatter/side scatter (FSC/SSC) gating followed by dead cell exclusion using 7-amino-actinomycin D (AAD). Numbers shown in each quadrant are the percentage of total gated cells.
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Figure 2: Cytokines alter the cell surface expression of CD16 and human CD14+ cells undergo a transitional stage of CD16 up-regulation in OC-promoting culture conditions. (a) Enriched human monocytes were cultured in osteoclast (OC)-promoting media (receptor activator of nuclear factor kappa-B ligand (RANKL) + macrophage colony-stimulating factor (M-CSF), pink line) or dendritic cell (DC)-promoting media (IL-4 + granulocyte-macrophage colony-stimulating factor (GM-CSF), green line). Freshly isolated monocytes (blue line) and the isotype control (black line) are also shown. Surface expression of CD16 was monitored by FACS analysis on [a] day 0, [b] day 1, and [c] day 3, respectively. (b) Human peripheral blood mononuclear cells (PBMC) were cultured in OC-promoting media (RANKL + M-CSF) and the cell surface expression of CD14 and CD16 was monitored on [a] day 0, [b] day 3, and [c] day 5. Data shown here are live cells after forward scatter/side scatter (FSC/SSC) gating followed by dead cell exclusion using 7-amino-actinomycin D (AAD). Numbers shown in each quadrant are the percentage of total gated cells.

Mentions: The cell surface expression of CD16 was monitored at two different time points after monocytes were purified from PBMC and cultured in plain, OC-promoting (RANKL + M-CSF), or DC-promoting (GM-CSF + IL-4) media (Figure 2(a)). Anti-CD16-PE, anti-CD14-allophycocyanin (APC) antibodies and 7-amino-actinomycin D (AAD) were included in our antibody staining panel. PBMC was first gated using forward scatter/side scatter (FSC/SSC) followed by dead cell exclusion by 7-AAD. Freshly isolated human monocytes are heterogeneous in regard to CD16 surface expression, and the majority of cells are CD16- (blue line in Figure 2(a)-a). After 24 hours in culture, the expression of CD16 increased in all culture conditions (Figure 2(a)-b). A significant polarization of CD16 cell surface expression was observed when cells were cultured for a longer period of time. By day three (Figure 2(a)-c), CD16 surface expression decreased on cells cultured in IL4+GM-CSF (green line in Figure 2(a)-c), but increased on cells cultured in RANKL+M-CSF (pink line in Figure 2(a)-c). Our results showed that CD16 upregulation occurred when monocytes were driven into OC but not DC lineage.


CD16 (FcRgammaIII) as a potential marker of osteoclast precursors in psoriatic arthritis.

Chiu YG, Shao T, Feng C, Mensah KA, Thullen M, Schwarz EM, Ritchlin CT - Arthritis Res. Ther. (2010)

Cytokines alter the cell surface expression of CD16 and human CD14+ cells undergo a transitional stage of CD16 up-regulation in OC-promoting culture conditions. (a) Enriched human monocytes were cultured in osteoclast (OC)-promoting media (receptor activator of nuclear factor kappa-B ligand (RANKL) + macrophage colony-stimulating factor (M-CSF), pink line) or dendritic cell (DC)-promoting media (IL-4 + granulocyte-macrophage colony-stimulating factor (GM-CSF), green line). Freshly isolated monocytes (blue line) and the isotype control (black line) are also shown. Surface expression of CD16 was monitored by FACS analysis on [a] day 0, [b] day 1, and [c] day 3, respectively. (b) Human peripheral blood mononuclear cells (PBMC) were cultured in OC-promoting media (RANKL + M-CSF) and the cell surface expression of CD14 and CD16 was monitored on [a] day 0, [b] day 3, and [c] day 5. Data shown here are live cells after forward scatter/side scatter (FSC/SSC) gating followed by dead cell exclusion using 7-amino-actinomycin D (AAD). Numbers shown in each quadrant are the percentage of total gated cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC2875642&req=5

Figure 2: Cytokines alter the cell surface expression of CD16 and human CD14+ cells undergo a transitional stage of CD16 up-regulation in OC-promoting culture conditions. (a) Enriched human monocytes were cultured in osteoclast (OC)-promoting media (receptor activator of nuclear factor kappa-B ligand (RANKL) + macrophage colony-stimulating factor (M-CSF), pink line) or dendritic cell (DC)-promoting media (IL-4 + granulocyte-macrophage colony-stimulating factor (GM-CSF), green line). Freshly isolated monocytes (blue line) and the isotype control (black line) are also shown. Surface expression of CD16 was monitored by FACS analysis on [a] day 0, [b] day 1, and [c] day 3, respectively. (b) Human peripheral blood mononuclear cells (PBMC) were cultured in OC-promoting media (RANKL + M-CSF) and the cell surface expression of CD14 and CD16 was monitored on [a] day 0, [b] day 3, and [c] day 5. Data shown here are live cells after forward scatter/side scatter (FSC/SSC) gating followed by dead cell exclusion using 7-amino-actinomycin D (AAD). Numbers shown in each quadrant are the percentage of total gated cells.
Mentions: The cell surface expression of CD16 was monitored at two different time points after monocytes were purified from PBMC and cultured in plain, OC-promoting (RANKL + M-CSF), or DC-promoting (GM-CSF + IL-4) media (Figure 2(a)). Anti-CD16-PE, anti-CD14-allophycocyanin (APC) antibodies and 7-amino-actinomycin D (AAD) were included in our antibody staining panel. PBMC was first gated using forward scatter/side scatter (FSC/SSC) followed by dead cell exclusion by 7-AAD. Freshly isolated human monocytes are heterogeneous in regard to CD16 surface expression, and the majority of cells are CD16- (blue line in Figure 2(a)-a). After 24 hours in culture, the expression of CD16 increased in all culture conditions (Figure 2(a)-b). A significant polarization of CD16 cell surface expression was observed when cells were cultured for a longer period of time. By day three (Figure 2(a)-c), CD16 surface expression decreased on cells cultured in IL4+GM-CSF (green line in Figure 2(a)-c), but increased on cells cultured in RANKL+M-CSF (pink line in Figure 2(a)-c). Our results showed that CD16 upregulation occurred when monocytes were driven into OC but not DC lineage.

Bottom Line: Exposure of cells to OC-promoting, but not DC-promoting media, was associated with CD16 up-regulation.An increased frequency of circulating CD14+CD16+ cells was noted in PsA compared to controls, and intermediate levels of CD16 may suggest a transitional state of OCP during osteoclastogenesis.Collectively, our data suggest that CD16 has the potential to serve as an OCP marker in inflammatory arthritis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Allergy/Immunology & Rheumatology Unit, University of Rochester Medical School, 601 Elmwood Avenue, Rochester, NY 14642, USA. grace_chiu@urmc.rochester.edu

ABSTRACT

Introduction: Psoriatic arthritis (PsA) is a chronic inflammatory arthritis characterized by bone erosion mediated by osteoclasts (OC). Our previous studies showed an elevated frequency of OC precursors (OCP) in PsA patients. Here, we examined if OC arise from CD16-positive monocytes in PsA.

Methods: Peripheral blood mononuclear cells (PBMC) or monocytes were isolated from human peripheral blood and sorted based on CD16 expression. Sorted cells were cultured alone or with bone wafers in the presence of receptor activator of nuclear factor kappa-B ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). Enumeration and bone erosion activity of OC were examined after culture. The effects of tumor necrosis factor-alpha (TNFalpha), OC-promoting (M-CSF plus RANKL), and dendritic cell (DC)-promoting (GM-CSF plus interleukin (IL)-4) cytokines on CD16 surface expression were examined by flow cytometry.

Results: PsA and psoriasis (Ps) subjects had a higher percentage of circulating inflammatory CD14+CD16+ cells than healthy controls (HC). Exposure of cells to OC-promoting, but not DC-promoting media, was associated with CD16 up-regulation. PBMC of Ps and PsA had a higher frequency of cells expressing intermediate levels of CD16. OC were mainly derived from CD16+ cells in PsA. Increased CD16 expression was associated with a higher bone erosion activity in PsA.

Conclusions: An increased frequency of circulating CD14+CD16+ cells was noted in PsA compared to controls, and intermediate levels of CD16 may suggest a transitional state of OCP during osteoclastogenesis. Intriguingly, TNFalpha blocked CD16 expression on a subset of CD14+ monocytes. Collectively, our data suggest that CD16 has the potential to serve as an OCP marker in inflammatory arthritis.

Show MeSH
Related in: MedlinePlus