The Journal of Experimental Medicine
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© The Rockefeller University Press, 0022-1007/1997/7/313/ $5.00
The Journal of Experimental Medicine, Volume 186, Number 2, July 21, 1997 313-323

Article

Human Peripheral Blood Eosinophils Express a Functional c-kit Receptor for Stem Cell Factor that Stimulates Very Late Antigen 4 (VLA-4)–mediated Cell Adhesion to Fibronectin and Vascular Cell Adhesion Molecule 1 (VCAM-1)

Qian Yuan*,||, K. Frank Austen*,||, Daniel S. Friend{ddagger}, Matthew Heidtman||, and Joshua A. Boyce*,§,||

From the * Department of Medicine, {ddagger} Department of Pathology, and § Department of Pediatrics, Harvard Medical School, the || Division of Rheumatology, Immunology, and Allergy, and the {ddagger}Department of Pathology, Brigham and Women's Hospital, Boston, Massachusetts 02115

We evaluated mature peripheral blood eosinophils for their expression of the surface tyrosine kinase, c-kit, the receptor for the stromal cell–derived cytokine, stem cell factor (SCF). Cytofluorographic analysis revealed that c-kit was expressed on the purified peripheral blood eosinophils from 8 of 8 donors (4 nonatopic and 4 atopic) (mean channel fluorescence intensity 2.0– 3.6-fold, average 2.8 ± 0.6-fold, greater than the negative control). The uniform and selective expression of c-kit by eosinophils was confirmed by immunohistochemical analysis of peripheral blood buffy coats. The functional integrity of c-kit was demonstrated by the capacity of 100 ng/ml (5 nM) of recombinant human (rh) SCF to increase eosinophil adhesion to 3, 10, and 30 µg/ml of immobilized FN40, a 40-kD chymotryptic fragment of plasma fibronectin, in 15 min by 7.7 ± 1.4-, 5.3 ± 3.3-, and 5.4 ± 0.2-fold, respectively, and their adhesion to 0.1, 0.5, and 1.0 µg/ml vascular cell adhesion molecule-1 (VCAM-1), by 12.7 ± 9.2-, 3.8 ± 2.5-, and 1.7 ± 0.6-fold, respectively. The SCF-stimulated adhesion occurred without concomitant changes in surface integrin expression, thereby indicating an avidity-based mechanism. rhSCF (100 ng/ml, 5 nM) was comparable to rh eotaxin (200 ng/ml, 24 nM) in stimulating adhesion. Cell adhesion to FN40 was completely inhibited with antibodies against the {alpha}4 and β1 integrin subunits, revealing that the SCF/c-kit adhesion effect was mediated by a single integrin heterodimer, very late antigen 4 (VLA-4). Thus, SCF represents a newly recognized stromal ligand for the activation of eosinophils for VLA-4–mediated adhesion, which could contribute to the exit of these cells from the blood, their tissue localization, and their prominence in inflammatory lesions.

Address correspondence to Qian Yuan, M.D., Ph.D., Harvard Medical School, Seeley G. Mudd Building, Room 618, 250 Longwood Avenue, Boston, MA 02115. Phone: 617-432-1995; FAX: 617-432-0979.

1Abbreviations used in this paper: BCECF-AM, 2',7'-bis-(2-carboxyethyl)- 5-(and 6)-carboxyfluorescein acetoxymethyl ester; BMMC, bone marrow–derived mast cells; CS-1, connecting segment-1; HSA, human serum albumin; MACS, magnetic cell separation; MFI, mean fluorescence intensity; rh, recombinant human; SCF, stem cell factor; VCAM, vascular cell adhesion molecule; VLA, very late antigen.


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