Characterization of a CSF-induced proliferating subpopulation of human peripheral blood monocytes by surface marker expression and cytokine production

Abstract

The phenotype of a subpopulation(s) of human monocytes which has been shown to proliferate in vitro in response to macrophage colonystimulating factor (M-CSF or CSF-1) and granulocyte-macrophage CSF (GM-CSF) is as yet unknown. To identify this proliferating subpopulation(s) we demonstrated first that DNA synthesis was occurring under culture conditions suitable for flow cytometric evaluation. Flow cytometric analysis of surface antigen expression identified that after 5 days of culture the proliferating subpopulation of monocytes expressed CD14, CD13, CD33, CD11b, CD11c, CD87, HLA-DR, CD45RO, and did not express CD86, CD34, CD80, CD4, CD16, and CD56. In addition, these proliferating monocytes (representing approximately 5% of total monocytes) were shown to produce the proinflammatory cytokines interleukin-6 and tumor necrosis factor ␣ in response to lipopolysaccharide stimulation. Further characterization and subsequent isolation of this subpopulation of monocytes may provide new and important information necessary to understand inflammatory diseases such as rheumatoid arthritis, where local proliferation at the site of inflammation may be a key factor contributing to the chronicity of the disease. J. Leukoc. Biol. 66: 953-960; 1999.

Measles Virus Glycoprotein-Pseudotyped Lentiviral Vectors Are Highly Superior to Vesicular Stomatitis Virus G Pseudotypes for Genetic Modification of Monocyte-Derived Dendritic Cells

“…3]). These results were similar for several donors and in agreement with recent findings suggesting that a small subset of human blood monocytes may be osteoclast precursors with proliferative properties (14,24,55). In addition, we confirmed that immature (day 4) and LPSmatured MDDCs (day 8) resided in the G 0 phase of the cell cycle (Fig.…”

Peripheral blood gene expression profiling in rheumatoid arthritis

“…Of the 52 genes that are overexpressed in RA PBMCs, 21 (40%) were in this 'monocyte-enriched' group. Thus, the presence of these genes may reflect in part the increased numbers 7,8 or activation of monocytes in RA PBMCs compared with controls (see below).…”

Stimulus-Dependent Requirement for Granulocyte-Macrophage Colony-Stimulating Factor in Inflammation

“…Brefeldin A (10 g/ml; Sigma-Aldrich) was added to the cultures for the final 2 h of LPS stimulation to limit cytokine secretion (29). Following FcR blocking, cells were stained for CD11b (Mac-1 ␣-chain; M1/70-allophycocyanin) as above, fixed in 4% paraformaldehyde, and permeabilized using 0.5% saponin and 2% FCS in PBS.…”