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Vol. 12, Issue 10, 3074-3086, October 2001
: Implications for
Leukocyte Transmigration
and
§
*Institute for Prevention of Cardiovascular Disease,
Ludwig-Maximilians-University, Munich, Germany 80336;
We investigated the role of H-Ras in chemokine-induced
integrin regulation in leukocytes. Stimulation of Jurkat
T cells with the CXC chemokine stromal cell-derived factor-1Department of Cardiovascular Molecular Medicine,
University Hospital, Aachen, Germany D-52074; and
Division of Rheumatology and Immunology, Brigham and
Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115
(SDF-1) resulted in a rapid increase in the phosphorylation, i.e.,
activation of extracellular signal receptor-activated kinase (ERK) but
not c-Jun NH2-terminal kinase or p38 kinase, and
phosphorylation of Akt, reflecting phosphatidylinositol
3-kinase (PI3-K) activation. Phosphorylation of ERK in Jurkat cells was
enhanced and attenuated by expression of dominant active (D12) or
inactive (N17) forms of H-Ras, respectively, while N17 H-Ras abrogated
SDF-1-induced Akt phosphorylation. SDF-1 triggered a transient
regulation of adhesion to intercellular adhesion molecule-1 (ICAM-1)
and vascular cell adhesion molecule-1 mediated by lymphocyte function
antigen-1 (LFA-1) and very late antigen-4 (VLA-4), respectively,
and a rapid increase in LFA-1 binding to soluble ICAM-1.Ig, which was
inhibited by D12 but not N17 H-Ras. Both D12 and N17 H-Ras abrogated
the regulation of LFA-1 but not VLA-4 avidity, and impaired
LFA-1-mediated transendothelial chemotaxis but not VLA-4-dependent
transmigration induced by SDF-1. Analysis of the mutant Jurkat J19
clone revealed LFA-1 with constitutively high affinity and reduced ERK
phosphorylation, which were partially restored by expression of active
H-Ras. Inhibition of PI3-K blocked the up-regulation of Jurkat cell
adhesion to ICAM-1 by SDF-1, whereas inhibition of mitogen-activated
protein kinase kinase impaired the subsequent down-regulation and
blocking both pathways abrogated LFA-1 regulation. Our data suggest
that inhibition of initial PI3-K activation by inactive H-Ras or
sustained activation of an inhibitory ERK pathway by active H-Ras
prevail to abolish LFA-1 regulation and transendothelial migration
induced by SDF-1 in leukocytes, establishing a complex and bimodal
involvement of H-Ras.
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