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The CD16(+) (FcgammaRIII(+)) subset of human monocytes preferentially becomes migratory dendritic cells in a model tissue setting.

Randolph GJ, Sanchez-Schmitz G, Liebman RM, Schäkel K - J. Exp. Med. (2002)

Bottom Line: These DCs migrate across endothelium in the ablumenal-to-lumenal direction (reverse transmigration), reminiscent of the migration into lymphatic vessels.CD16 was not functionally required for reverse transmigration, but promoted cell survival when yeast particles (zymosan) were present as a maturation stimulus in the subendothelial matrix.We propose that CD16(+) monocytes may contribute significantly to precursors for DCs that transiently survey tissues and migrate to lymph nodes via afferent lymphatic vessels.

View Article: PubMed Central - PubMed

Affiliation: The Carl C. Icahn Institute for Gene Therapy and Molecular Medicine, Mt. Sinai School of Medicine, 1425 Madison Avenue, New York, NY 10029, USA. gwendalyn.randolph@mssm.edu

ABSTRACT
Much remains to be learned about the physiologic events that promote monocytes to become lymph-homing dendritic cells (DCs). In a model of transendothelial trafficking, some monocytes become DCs in response to endogenous signals. These DCs migrate across endothelium in the ablumenal-to-lumenal direction (reverse transmigration), reminiscent of the migration into lymphatic vessels. Here we show that the subpopulation of monocytes that expresses CD16 (Fcgamma receptor III) is predisposed to become migratory DCs. The vast majority of cells derived from CD16(+) monocytes reverse transmigrated, and their presence was associated with migratory cells expressing high levels of CD86 and human histocompatibility leukocyte antigen (HLA)-DR, and robust capacity to induce allogeneic T cell proliferation. A minority of CD16(-) monocytes reverse transmigrated, and these cells stimulated T cell proliferation less efficiently. CD16 was not functionally required for reverse transmigration, but promoted cell survival when yeast particles (zymosan) were present as a maturation stimulus in the subendothelial matrix. The cell surface phenotype and migratory characteristics of CD16(+) monocytes were inducible in CD16(-) monocytes by preincubation with TGFbeta1. We propose that CD16(+) monocytes may contribute significantly to precursors for DCs that transiently survey tissues and migrate to lymph nodes via afferent lymphatic vessels.

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Cell surface markers distinguish CD16+ and CD16−HLA-DR+ PBMCs. Freshly isolated PBMCs were depleted of CD56+ NK cells. Two-color flow cytometry was conducted with a gate set to exclude smaller PBMCs of the lymphocyte lineage. Expression of macrophage-associated markers CD14, CD36, and CD64 or DC-associated markers HLA-DR, HLA-DP, and CD86 (all x-axis) were examined. Cells were counterstained with a mAb to CD16 (y-axis). Some cells were stained with M-DC8 mAb to identify a subset of CD16+ cells (reference 11). Quadrant markers (shown in CD14 and CD123 panels) are positioned according to the level of fluorescence observed in cells stained with nonbinding isotype-matched control mAbs (lower left quadrant, negative staining). The phenotype illustrated was similarly observed among four donors examined.
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fig1: Cell surface markers distinguish CD16+ and CD16−HLA-DR+ PBMCs. Freshly isolated PBMCs were depleted of CD56+ NK cells. Two-color flow cytometry was conducted with a gate set to exclude smaller PBMCs of the lymphocyte lineage. Expression of macrophage-associated markers CD14, CD36, and CD64 or DC-associated markers HLA-DR, HLA-DP, and CD86 (all x-axis) were examined. Cells were counterstained with a mAb to CD16 (y-axis). Some cells were stained with M-DC8 mAb to identify a subset of CD16+ cells (reference 11). Quadrant markers (shown in CD14 and CD123 panels) are positioned according to the level of fluorescence observed in cells stained with nonbinding isotype-matched control mAbs (lower left quadrant, negative staining). The phenotype illustrated was similarly observed among four donors examined.

Mentions: The HLA-DR+ fraction of PBMCs that lack characteristics of B lymphocytes represent ∼20% of PBMCs and contain several subpopulations that collectively comprise potential precursors for antigen-presenting DCs. These HLA-DR+ cells can be subdivided into several distinct populations, as shown in Fig. 1 : CD14− nonmonocytic DCs that contain a very small fraction of CD123+ plasmacytoid cells, and CD14+ monocytes that can be divided into CD16− and CD16+ fractions. The CD14+CD16+ monocytes can be further subdivided into cells that do or do not express the DC subset marker M-DC8 (14, 15). The CD16+ subpopulation of monocytes expresses only an intermediate level of CD14 relative to CD16− monocytes (12; Fig. 1). These cells exhibit reduced levels of other markers associated with macrophage activities, including CD36 and the high affinity Fcγ receptor CD64 (Fig. 1). In contrast, CD16+ monocytes exhibit somewhat elevated levels of accessory molecules, including CD86, HLA-DR (16), and HLA-DP (Fig. 1). These data indicate that CD16+ monocytes more closely resemble DCs than do CD16− monocytes.


The CD16(+) (FcgammaRIII(+)) subset of human monocytes preferentially becomes migratory dendritic cells in a model tissue setting.

Randolph GJ, Sanchez-Schmitz G, Liebman RM, Schäkel K - J. Exp. Med. (2002)

Cell surface markers distinguish CD16+ and CD16−HLA-DR+ PBMCs. Freshly isolated PBMCs were depleted of CD56+ NK cells. Two-color flow cytometry was conducted with a gate set to exclude smaller PBMCs of the lymphocyte lineage. Expression of macrophage-associated markers CD14, CD36, and CD64 or DC-associated markers HLA-DR, HLA-DP, and CD86 (all x-axis) were examined. Cells were counterstained with a mAb to CD16 (y-axis). Some cells were stained with M-DC8 mAb to identify a subset of CD16+ cells (reference 11). Quadrant markers (shown in CD14 and CD123 panels) are positioned according to the level of fluorescence observed in cells stained with nonbinding isotype-matched control mAbs (lower left quadrant, negative staining). The phenotype illustrated was similarly observed among four donors examined.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2196052&req=5

fig1: Cell surface markers distinguish CD16+ and CD16−HLA-DR+ PBMCs. Freshly isolated PBMCs were depleted of CD56+ NK cells. Two-color flow cytometry was conducted with a gate set to exclude smaller PBMCs of the lymphocyte lineage. Expression of macrophage-associated markers CD14, CD36, and CD64 or DC-associated markers HLA-DR, HLA-DP, and CD86 (all x-axis) were examined. Cells were counterstained with a mAb to CD16 (y-axis). Some cells were stained with M-DC8 mAb to identify a subset of CD16+ cells (reference 11). Quadrant markers (shown in CD14 and CD123 panels) are positioned according to the level of fluorescence observed in cells stained with nonbinding isotype-matched control mAbs (lower left quadrant, negative staining). The phenotype illustrated was similarly observed among four donors examined.
Mentions: The HLA-DR+ fraction of PBMCs that lack characteristics of B lymphocytes represent ∼20% of PBMCs and contain several subpopulations that collectively comprise potential precursors for antigen-presenting DCs. These HLA-DR+ cells can be subdivided into several distinct populations, as shown in Fig. 1 : CD14− nonmonocytic DCs that contain a very small fraction of CD123+ plasmacytoid cells, and CD14+ monocytes that can be divided into CD16− and CD16+ fractions. The CD14+CD16+ monocytes can be further subdivided into cells that do or do not express the DC subset marker M-DC8 (14, 15). The CD16+ subpopulation of monocytes expresses only an intermediate level of CD14 relative to CD16− monocytes (12; Fig. 1). These cells exhibit reduced levels of other markers associated with macrophage activities, including CD36 and the high affinity Fcγ receptor CD64 (Fig. 1). In contrast, CD16+ monocytes exhibit somewhat elevated levels of accessory molecules, including CD86, HLA-DR (16), and HLA-DP (Fig. 1). These data indicate that CD16+ monocytes more closely resemble DCs than do CD16− monocytes.

Bottom Line: These DCs migrate across endothelium in the ablumenal-to-lumenal direction (reverse transmigration), reminiscent of the migration into lymphatic vessels.CD16 was not functionally required for reverse transmigration, but promoted cell survival when yeast particles (zymosan) were present as a maturation stimulus in the subendothelial matrix.We propose that CD16(+) monocytes may contribute significantly to precursors for DCs that transiently survey tissues and migrate to lymph nodes via afferent lymphatic vessels.

View Article: PubMed Central - PubMed

Affiliation: The Carl C. Icahn Institute for Gene Therapy and Molecular Medicine, Mt. Sinai School of Medicine, 1425 Madison Avenue, New York, NY 10029, USA. gwendalyn.randolph@mssm.edu

ABSTRACT
Much remains to be learned about the physiologic events that promote monocytes to become lymph-homing dendritic cells (DCs). In a model of transendothelial trafficking, some monocytes become DCs in response to endogenous signals. These DCs migrate across endothelium in the ablumenal-to-lumenal direction (reverse transmigration), reminiscent of the migration into lymphatic vessels. Here we show that the subpopulation of monocytes that expresses CD16 (Fcgamma receptor III) is predisposed to become migratory DCs. The vast majority of cells derived from CD16(+) monocytes reverse transmigrated, and their presence was associated with migratory cells expressing high levels of CD86 and human histocompatibility leukocyte antigen (HLA)-DR, and robust capacity to induce allogeneic T cell proliferation. A minority of CD16(-) monocytes reverse transmigrated, and these cells stimulated T cell proliferation less efficiently. CD16 was not functionally required for reverse transmigration, but promoted cell survival when yeast particles (zymosan) were present as a maturation stimulus in the subendothelial matrix. The cell surface phenotype and migratory characteristics of CD16(+) monocytes were inducible in CD16(-) monocytes by preincubation with TGFbeta1. We propose that CD16(+) monocytes may contribute significantly to precursors for DCs that transiently survey tissues and migrate to lymph nodes via afferent lymphatic vessels.

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