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1 Medical Research Service,
Seattle Veterans Affairs Medical Center, and Division of Pulmonary
and Critical Care Medicine, Department of Medicine, University of
Washington School of Medicine, Seattle, Washington 98108;
2 Division of Pulmonary and
Critical Care Medicine,
The C-X-C chemokines are a structurally related
and functionally redundant family of proteins with neutrophil
chemotactic activity. Many of the C-X-C chemokines are produced by
endotoxin-stimulated alveolar macrophages (AMs), but knowledge of their
relative quantities and their relative contributions to the total
chemotactic activity released from these cells is incomplete. Human AMs
were stimulated with or without Escherichia
coli endotoxin for 2, 4, 8, and 24 h. The mRNA
sequences of interleukin (IL)-8, the 78-amino acid epithelial
cell-derived neutrophil activator (ENA-78), growth-related protein
(GRO)
, GRO
, and GRO
were cloned by PCR and
identified by sequence analysis. The relative mRNA quantities were
compared by Northern analysis, and IL-8 was found to predominate.
Similarly, IL-8 protein concentrations in the cell supernatants were
consistently higher than either the ENA-78 or GRO concentration, and by
24 h, IL-8 concentrations were 10-fold higher than those of the other C-X-C chemokines. Blocking polyclonal antibodies to IL-8 substantially reduced the chemotactic activity in the AM supernatants, whereas antibodies to ENA-78 and GRO had little or no effect. We conclude that
IL-8 is the predominant C-X-C chemokine and the dominant neutrophil
chemoattractant accumulating in 24-h supernatants of lipopolysaccharide-stimulated human AMs. These studies provide insight
into potentially effective strategies of interrupting AM-derived
inflammatory signals in the lungs.
chemotaxis; monocytes; neutrophils; lung
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