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 Blood, 1 November 2004, Vol. 104, No. 9, pp. 2818-2824.
Prepublished online as a Blood First Edition Paper on July 8, 2004; DOI 10.1182/blood-2003-12-4402.

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IMMUNOBIOLOGY

Paxillin selectively associates with constitutive and chemoattractant-induced high-affinity {alpha}4{beta}1 integrins: implications for integrin signaling

Sharon J. Hyduk, Jiwon Oh, Haiyan Xiao, Mian Chen, and Myron I. Cybulsky

From the Toronto General Research Institute and Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada.

Leukocyte {alpha}4{beta}1 integrins regulate hematopoietic and lymphoid development, as well as the emigration of circulating cells to sites of inflammation. Because vascular cell adhesion molecule-1 (VCAM-1) binding to high-affinity {alpha}4{beta}1 is stable, these integrins can be detected and selectively precipitated from cell lysates using VCAM-1/Fc. With this approach, high-affinity {alpha}4{beta}1 integrin expression was demonstrated on lymphocytes in the bone marrow, thymus, spleen, and the peritoneal cavity of normal mice, but not in peripheral lymph nodes. Immature lymphocytes preferentially expressed high-affinity {alpha}4{beta}1 in the bone marrow and thymus. Paxillin is a cytoplasmic adaptor molecule that can bind to the {alpha}4 tail and initiate signaling. Paxillin was associated selectively with high-affinity integrins that were isolated from human Jurkat T cells or from murine tissues, and blotting with a phospho-specific antibody demonstrated that Ser988 in the {alpha}4 cytoplasmic tail was dephosphorylated in high-affinity but not low-affinity integrins. A rapid and transient {alpha}4{beta}1 affinity up-regulation in formyl peptide receptor-transfected U937 cells stimulated with N-formyl-methyonyl-leucyl-phenylalanine (fMLP) correlated temporally with induced paxillin binding to {alpha}4 integrins. These data suggest that ligand binding to high-affinity {alpha}4{beta}1 integrins may initiate outside-in signaling cascades through paxillin that regulate leukocyte maturation and emigration. (Blood. 2004;104:2818-2824)


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