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Departments of
* Cancer Biology and
Vascular Biology, La Jolla Institute for Molecular Medicine, San Diego, CA 92121
In leukocytes, C3a and C5a cause chemotaxis in a
Gi-dependent, pertussis toxin (PT)-sensitive fashion.
Because we found that HUVECs and immortalized human dermal
microvascular endothelial cells express small numbers of C3aRs and
C5aRs, we asked what the function of these receptors was on these
cells. Activation of the C3aR caused transient formation of actin
stress fibers, which was not PT-sensitive, but depended on rho
activation implying coupling to G
12 or
G13. Activation of the C5aR caused a delayed and
sustained cytoskeletal response, which was blocked by PT, and resulted
in cell retraction, increased paracellular permeability, and
facilitated eosinophil transmigration. C5a, but not C3a, was
chemotactic for human immortalized dermal microvascular endothelial
cells. The response to C5a was blocked by inhibitors of
phosphatidylinositol-3-kinase, src kinase, and of the epidermal growth
factor (EGF) receptor (EGFR) as well as by neutralizing Abs against the
EGFR and heparin-binding EGF-like factor. Furthermore, immune
precipitations showed that the EGFR was phosphorylated following
stimulation with C5a. The C5aR in endothelial cells thus uses a
signaling cascade–transactivation of the EGFR–that does not exist in
leukocytes, while the C3aR couples to a different G protein, presumably
G12/13.
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