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1 Department of Internal Medicine and 2 Department of Pathology, Justus-Liebig-University, Giessen 35392, Germany
The evaluation of
monocytes recruited into the alveolar space under both physiological
and inflammatory conditions is hampered by difficulties in
discriminating these cells from resident alveolar macrophages (rAMs).
Using the intravenous injected fluorescent dye PKH26, which accumulated
in rAMs without labeling blood leukocytes, we developed a technique
that permits the identification, isolation, and functional analysis of
monocytes recruited into lung alveoli of mice. Alveolar deposition of
murine JE, the homologue of human monocyte chemoattractant protein
(MCP)-1 (JE/MCP-1), in mice provoked an alveolar influx of monocytes
that were recovered by bronchoalveolar lavage and separated from
PKH26-stained rAMs by flow cytometry. Alveolar recruited monocytes
showed a blood monocytic phenotype as assessed by cell surface
expression of F4/80, CD11a, CD11b, CD18, CD49d, and CD62L. In contrast,
CD14 was markedly upregulated on alveolar recruited monocytes together
with increased tumor necrosis factor-
message, discriminating this
monocyte population from peripheral blood monocytes and rAMs. Thus
monocytes recruited into the alveolar air space of mice in response to
JE/MCP-1 keep phenotypic features of blood monocytes but upregulate
CD14 and are "primed" for enhanced responsiveness to endotoxin with
increased cytokine expression.
monocyte chemoattractant protein-1; alveolar macrophage; fluorescent dye; flow cytometry
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