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Department of Immunobiology, DNAX Research Institute, Palo Alto, CA 94304
The chemokine RANTES is a potent agonist of T cell activation. In
an investigation of signal-transduction events activated by this
chemokine, we have shown that RANTES stimulates dose-dependent
phospholipase D (PLD) activity in Jurkat cells. Equilibrium-binding
analyses using 125I-labeled RANTES indicated the
presence of a receptor for RANTES on these cells, which has a
Kd of 0.1 nM, is expressed at approximately 600
sites per cell, and a binding specificity that was not comparable with
that of any of the known chemokine receptors, since
125I-labeled RANTES was displaced by
macrophage-inflammatory protein-1ß (but not macrophage-inflammatory
protein-1
), monocyte-chemotactic protein-1 (MCP-1), MCP-3, MCP-4,
and eotaxin. RANTES-induced PLD activation was augmented by GTP
S,
but not GDPßS, and inhibited by the protein kinase C inhibitor
bisindolylmaleimide, as well as the fungal metabolite brefeldin A, and
C3 exoenzyme (Clostridium botulinum),
implicating the activation of RhoA. RANTES also induced GTP-GDP
exchange of immunoprecipitated RhoA. RANTES-stimulated PLD activity was
dependent on an ADP-ribosylation factor(s), as assessed by inhibition
studies using a synthetic inhibitory peptide of the N-terminal 16 amino
acids of ADP-ribosylation factor 1. These studies indicate the
potential existence of a novel receptor-mediated mechanism for
activation of T cells by the chemokine RANTES.
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