Comparative Analysis of the Morphological, Cytochemical, Immunophenotypical, and Functional Characteristics of Normal Human Peripheral Blood Lineage/CD16/HLA-DR/CD14 Cells, CD14 Monocytes, and CD16 Dendritic Cells |
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Authors: | Julia Almeida, Clara Bueno, Ma Carmen Alguer , Ma Luz Sanchez, Ma de Santiago, Luis Escribano, Beatriz Dí az-Agustí n, Jose Miguel Vaquero, F. Javier Laso, Jesus F. San Miguel,Alberto Orfao |
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Affiliation: | a Servicio General de Citometría y Departamento de Medicina, Universidad de Salamanca, Salamanca, Hospital Ramón y Cajal, Madrid;b Centro de Investigación del Cáncer, Universidad de Salamanca, Salamanca, Hospital Ramón y Cajal, Madrid;c Universidad de Salamanca, Salamanca, Servicio de Hematología, Hospital Ramón y Cajal, Madrid;d Servicio de Medicina Interna, Hospital Universitario de Salamanca, 37007, Salamanca, Spain;e Servicio de Hematología, Hospital Universitario de Salamanca, 37007, Salamanca, Spain |
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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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Keywords: | human dendritic cell subsets flow cytometry immunophenotype phagocytic/oxidative burst activity cytokines |
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