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The Journal of Immunology, 2001, 166: 4115-4123.
Copyright © 2001 by The American Association of Immunologists

Resting Murine Neutrophils Express Functional {alpha}4 Integrins that Signal Through Src Family Kinases1

Shalini Pereira, Meijuan Zhou, Attila Mócsai and Clifford Lowell2

Department of Laboratory Medicine, University of California, San Francisco, CA 94143

There is mounting evidence that {alpha}4 (CD49d) integrins are involved in neutrophil recruitment and function during inflammatory responses. We report that all resting murine neutrophils derived from bone marrow or peripheral blood express easily detectable levels of {alpha}4 integrins on their surface. These {alpha}4 integrins were functional, as demonstrated by stimulation of respiratory burst when neutrophils adhered to surfaces coated with the murine vascular cell adhesion molecule-1 (mVCAM-1). Adhesion occurred via {alpha}4 integrins, as preincubation of neutrophils with an anti-{alpha}4-specific Ab inhibited attachment to mVCAM-1. Direct cross-linking of the {alpha}4 integrin subunit by surface-bound mAbs also elicited superoxide release and release of the secondary granule marker, lactoferrin. The functional responses that occurred downstream of {alpha}4 integrin cross-linking required signaling by Src family kinases. Neutrophils derived from hck-/-fgr-/-lyn-/- triple-knockout or hck-/-fgr-/- double-knockout mice failed to undergo respiratory burst when plated on mVCAM-1. Triple mutant neutrophils were also defective in release of both superoxide and lactoferrin when plated on surfaces coated with mAbs directed against {alpha}4. Correlated with impaired {alpha}4-induced functional responses, triple-mutant neutrophils also failed to spread and tightly adhere to anti-{alpha}4 mAb-coated surfaces. This is the first direct evidence that functional {alpha}4 integrins are expressed by murine PMNs, and that these surface molecules can mediate cellular responses such as tight adhesion, spreading, sustained respiratory burst, and specific granule release in vitro. Moreover the {alpha}4 integrins, like all other integrins tested, use the Src family kinases to transduce intracellular signals.




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