Elsevier

Clinical Immunology

Volume 100, Issue 3, September 2001, Pages 325-338
Clinical Immunology

Regular Article
Comparative Analysis of the Morphological, Cytochemical, Immunophenotypical, and Functional Characteristics of Normal Human Peripheral Blood Lineage/CD16+/HLA-DR+/CD14−/lo Cells, CD14+ Monocytes, and CD16 Dendritic Cells

https://doi.org/10.1006/clim.2001.5072Get rights and content

Abstract

Human peripheral blood (PB) CD14lo/HLA-DR+ cells were initially described as a subset of mature monocytes. Recently, it has been suggested that these represent a part of a new subset of dendritic cells (DC), characterized by the coexpression of MDC-8/HLA-DR/CD16. The aim of the present paper was to analyze the morphological, cytochemical, phenotypical, and functional characteristics of PB CD16+/HLA-DR+ cells compared to both PB CD14+ monocytes and CD16 DC. In contrast to CD14+ monocytes, purified CD16+/HLA-DR+ cells displayed cytoplasmic veils and lacked cytoplasmic myeloperoxidase and α-naphthyl acetate esterase. Normal human PB CD16+/HLA-DR+ cells also displayed phenotypic characteristics different from those of CD14+ monocytes: they lacked the CD64 Fcγ receptor, showed lower levels of CD32, and expressed higher amounts of CD16 compared to CD14+ monocytes. They also displayed a different pattern of expression of other antigens, including CD14, HLA-DR, CD45RA, CD45RO, complement receptors and complement regulatory surface proteins, adhesion and costimulatory molecules, and cytokine receptors, among others. When compared to CD16 DC, CD16+/HLA-DR+ cells showed reactivity for CD16, dim positivity for CD14, higher expression of both Ig- and complement-receptors and lower reactivity for HLA-DR, adhesion, and costimulatory molecules (with the exception of CD86). The CD16+/HLA-DR+ cell subset displayed a higher Ig/complement-mediated phagocytic/oxidative activity than CD16 DC, although this activity was significantly lower than that of mature monocytes. Regarding cytokine production at the single cell level, LPS plus IFN-γ-stimulated PB CD16+/HLA-DR+ cells produced significant amounts of IL1β, IL6, IL12, TNFα, and IL8; however, the percentage of cytokine-producing cells and the amount of cytokine/cell were lower in CD16+/HLA-DR+ cells than in CD14+ monocytes. In addition, upon comparing CD16+/HLA-DR+ cells with CD33+++/CD16 DC, we found that the percentage of cytokine-producing cells and the amount of cytokine/cell were significantly different in both cell subsets. In summary, our results show that CD16+/HLA-DR+ cells clearly display different morphologic, cytochemical, immunophenotypical, and functional characteristics compared to both mature monocytes and CD16 DC. Interestingly, these cells are more frequent than other DC in normal human adult PB and cord blood samples, while they are less represented in normal bone marrow.

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