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1
*
Department of Trauma Surgery, University of Freiburg Medical School, Freiburg/Breisgau, Germany; and
Department of Pathology, University of Michigan Medical School, Ann Arbor, MI 48109
Under a variety of conditions, alveolar macrophages can generate
early response cytokines (TNF-
, IL-1), complement components, and
chemotactic cytokines (chemokines). In the current studies, we
determined the requirements for TNF- and the complement activation
product C5a in chemokine production in vitro and in vivo. Two rat CXC
chemokines (macrophage inflammatory protein (MIP)-2 and
cytokine-induced neutrophil chemoattractant (CINC)) as well as three
rat CC chemokines (MIP-1, MIP-1ß, and monocyte chemoattractant
protein (MCP)-1) were investigated. Chemokine generation in vitro was
studied in rat alveolar macrophages stimulated with IgG immune
complexes in the absence or presence of Abs to TNF- or C5a. The rat
lung injury model induced by IgG immune complex deposition was employed
for in vivo studies. Abs to TNF- or C5a were administered
intratracheally or i.v., and effects on chemokine levels in
bronchoalveolar lavage fluids were quantitated by ELISA. Both in vitro
and in vivo studies demonstrated the requirements for TNF- and C5a
for full generation of CXC and CC chemokines. In vitro and in vivo
blockade of TNF- or C5a resulted in significantly reduced production
of chemokines. Supernatant fluids from in vitro-stimulated macrophages
revealed by Western blot analysis the presence of
C5a/C5adesArg, indicating intrinsic generation of
C5a/C5adesArg by alveolar macrophages and explaining the
higher efficiency of intratracheal vs i.v. blockade of C5a in reducing
chemokine production. These results underscore the central role of both
TNF- and C5a, which appear to function as autocrine activators to
promote CXC and CC chemokine generation by alveolar
macrophages.
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