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Characterization of the CD14++CD16+ monocyte population in human bone marrow.

Mandl M, Schmitz S, Weber C, Hristov M - PLoS ONE (2014)

Bottom Line: These cells express essential monocyte-related antigens and chemokine receptors such as CD11a, CD18, CD44, HLA-DR, Ccr2, Ccr5, Cx3cr1, Cxcr2 and Cxcr4.Notably, the expression of Ccr2 was inducible during culture.Furthermore, sorted CD14(++)CD16(+) bone marrow cells show typical macrophage morphology, phagocytic activity, angiogenic features and generation of intracellular oxygen species.

View Article: PubMed Central - PubMed

Affiliation: Institute for Cardiovascular Prevention, Ludwig-Maximilians-University (LMU), Munich, Germany.

ABSTRACT
Numerous studies have divided blood monocytes according to their expression of the surface markers CD14 and CD16 into following subsets: classical CD14(++)CD16(-), intermediate CD14(++)CD16(+) and nonclassical CD14(+)CD16(++) monocytes. These subsets differ in phenotype and function and are further correlated to cardiovascular disease, inflammation and cancer. However, the CD14/CD16 nature of resident monocytes in human bone marrow remains largely unknown. In the present study, we identified a major population of CD14(++)CD16(+) monocytes by using cryopreserved bone marrow mononuclear cells from healthy donors. These cells express essential monocyte-related antigens and chemokine receptors such as CD11a, CD18, CD44, HLA-DR, Ccr2, Ccr5, Cx3cr1, Cxcr2 and Cxcr4. Notably, the expression of Ccr2 was inducible during culture. Furthermore, sorted CD14(++)CD16(+) bone marrow cells show typical macrophage morphology, phagocytic activity, angiogenic features and generation of intracellular oxygen species. Side-by-side comparison of the chemokine receptor profile with unpaired blood samples also demonstrated that these rather premature medullar monocytes mainly match the phenotype of intermediate and partially of (non)classical monocytes. Together, human monocytes obviously acquire their definitive CD14/CD16 signature in the bloodstream and the medullar monocytes probably transform into CD14(++)CD16- and CD14(+)CD16(++) subsets which appear enriched in the periphery.

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Characterization of CD14++CD16+ bone marrow monocytes by flow cytometry.(A) Representative contour plot of pooled BMC sample from three single donors showing a clear separation of two populations: CD14++CD16+ (Q1) and CD14−CD16++ (Q4). Gating was performed first in FSC/SSC dot plot following gating on CD45+ events in a SSC/CD45 dot plot. Finally, the expression of CD14 and CD16 was evaluated inside the CD45+ population. (B,C) Expression of monocyte-related antigens and representative histogram overlays with isotype control (gray filled) of crucial chemokine receptors on CD14++CD16+ BMCs as analyzed by flow cytometry. Data are from 3 to 6 individual donors.
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pone-0112140-g001: Characterization of CD14++CD16+ bone marrow monocytes by flow cytometry.(A) Representative contour plot of pooled BMC sample from three single donors showing a clear separation of two populations: CD14++CD16+ (Q1) and CD14−CD16++ (Q4). Gating was performed first in FSC/SSC dot plot following gating on CD45+ events in a SSC/CD45 dot plot. Finally, the expression of CD14 and CD16 was evaluated inside the CD45+ population. (B,C) Expression of monocyte-related antigens and representative histogram overlays with isotype control (gray filled) of crucial chemokine receptors on CD14++CD16+ BMCs as analyzed by flow cytometry. Data are from 3 to 6 individual donors.

Mentions: Flow cytometry analysis of thawed human BMCs after staining for CD14, CD16 and CD45 showed clear separation of two scatter populations (Fig. 1A): a larger CD14++ population (Q1: 17.6±3.0% of CD45+ cells) with low-to-intermediate expression of CD16 (CD16+) and a smaller CD14−CD16++ population (Q4: 6.1±1.6% of CD45+ cells). The CD14−CD16++ cells in Q4 were also CD15−CD56++ thus probably referring to NK and NK-T cells (data not shown).


Characterization of the CD14++CD16+ monocyte population in human bone marrow.

Mandl M, Schmitz S, Weber C, Hristov M - PLoS ONE (2014)

Characterization of CD14++CD16+ bone marrow monocytes by flow cytometry.(A) Representative contour plot of pooled BMC sample from three single donors showing a clear separation of two populations: CD14++CD16+ (Q1) and CD14−CD16++ (Q4). Gating was performed first in FSC/SSC dot plot following gating on CD45+ events in a SSC/CD45 dot plot. Finally, the expression of CD14 and CD16 was evaluated inside the CD45+ population. (B,C) Expression of monocyte-related antigens and representative histogram overlays with isotype control (gray filled) of crucial chemokine receptors on CD14++CD16+ BMCs as analyzed by flow cytometry. Data are from 3 to 6 individual donors.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4219836&req=5

pone-0112140-g001: Characterization of CD14++CD16+ bone marrow monocytes by flow cytometry.(A) Representative contour plot of pooled BMC sample from three single donors showing a clear separation of two populations: CD14++CD16+ (Q1) and CD14−CD16++ (Q4). Gating was performed first in FSC/SSC dot plot following gating on CD45+ events in a SSC/CD45 dot plot. Finally, the expression of CD14 and CD16 was evaluated inside the CD45+ population. (B,C) Expression of monocyte-related antigens and representative histogram overlays with isotype control (gray filled) of crucial chemokine receptors on CD14++CD16+ BMCs as analyzed by flow cytometry. Data are from 3 to 6 individual donors.
Mentions: Flow cytometry analysis of thawed human BMCs after staining for CD14, CD16 and CD45 showed clear separation of two scatter populations (Fig. 1A): a larger CD14++ population (Q1: 17.6±3.0% of CD45+ cells) with low-to-intermediate expression of CD16 (CD16+) and a smaller CD14−CD16++ population (Q4: 6.1±1.6% of CD45+ cells). The CD14−CD16++ cells in Q4 were also CD15−CD56++ thus probably referring to NK and NK-T cells (data not shown).

Bottom Line: These cells express essential monocyte-related antigens and chemokine receptors such as CD11a, CD18, CD44, HLA-DR, Ccr2, Ccr5, Cx3cr1, Cxcr2 and Cxcr4.Notably, the expression of Ccr2 was inducible during culture.Furthermore, sorted CD14(++)CD16(+) bone marrow cells show typical macrophage morphology, phagocytic activity, angiogenic features and generation of intracellular oxygen species.

View Article: PubMed Central - PubMed

Affiliation: Institute for Cardiovascular Prevention, Ludwig-Maximilians-University (LMU), Munich, Germany.

ABSTRACT
Numerous studies have divided blood monocytes according to their expression of the surface markers CD14 and CD16 into following subsets: classical CD14(++)CD16(-), intermediate CD14(++)CD16(+) and nonclassical CD14(+)CD16(++) monocytes. These subsets differ in phenotype and function and are further correlated to cardiovascular disease, inflammation and cancer. However, the CD14/CD16 nature of resident monocytes in human bone marrow remains largely unknown. In the present study, we identified a major population of CD14(++)CD16(+) monocytes by using cryopreserved bone marrow mononuclear cells from healthy donors. These cells express essential monocyte-related antigens and chemokine receptors such as CD11a, CD18, CD44, HLA-DR, Ccr2, Ccr5, Cx3cr1, Cxcr2 and Cxcr4. Notably, the expression of Ccr2 was inducible during culture. Furthermore, sorted CD14(++)CD16(+) bone marrow cells show typical macrophage morphology, phagocytic activity, angiogenic features and generation of intracellular oxygen species. Side-by-side comparison of the chemokine receptor profile with unpaired blood samples also demonstrated that these rather premature medullar monocytes mainly match the phenotype of intermediate and partially of (non)classical monocytes. Together, human monocytes obviously acquire their definitive CD14/CD16 signature in the bloodstream and the medullar monocytes probably transform into CD14(++)CD16- and CD14(+)CD16(++) subsets which appear enriched in the periphery.

Show MeSH
Related in: MedlinePlus