Background: Transmembrane 4 superfamily (TM4SF) molecules are exclusively found on hematopoietic cells. Several members of the TM4SF are reported to be associated with other cell surface molecules, including integrins, and might participate in signal transduction, but little is known about their role on eosinophils. In the present study, we determined the expression and function of TM4SF molecules on human eosinophils. Methods: Surface expression of TM4SF molecules on purified peripheral blood eosinophils was examined using indirect immunofluorescence and flow cytometry. Purified eosinophils were incubated with anti-TM4SF monoclonal antibodies (mAbs) for up to 24 h. Eosinophil activation was evaluated by measuring eosinophil homotypic aggregation as well as changes in surface expression of CD11b or CD62L by flow cytometry. Results: Freshly isolated eosinophils expressed CD9, CD37, CD53, CD63 and CD81. Incubation with anti-CD9 mAb but not with anti-CD37, CD53, CD63 or CD81 mAb induced significant eosinophil homotypic aggregation. Incubation with any of the anti-TM4SF mAb for 30 min failed to alter the expression of either CD11b or CD62L on eosinophils. In contrast, the expression of CD11b was significantly enhanced after 24 h of incubation with anti-CD53 mAb, while the expression of CD62L was significantly reduced with anti-CD81 mAb. Conclusions: Cross-linking of some surface TM4SF molecules induced significant eosinophil homotypic aggregation, upregulation of CD11b expression, or CD62L shedding, consistent with activation of eosinophils. Our data suggest that several TM4SF molecules are functionally expressed on human eosinophils, and therefore might participate in allergic inflammation.

1.
Gleich GJ: The eosinophil and bronchial asthma: Current understanding. J Allergy Clin Immunol 1990;85:422–436.
2.
Silberstein DS, David JR: The regulation of human eosinophil function by cytokines. Immunol Today 1987;8:380–385.
3.
Hirai K, Miyamasu M, Takaishi T, Morita Y: Regulation of the function of eosinophils and basophils. Crit Rev Immunol 1997;17:325–352.
4.
Springer TA: Adhesion receptors of the immune system. Nature 1990;346:425–434.
5.
Weber C, Katayama J, Springer TA: Differential regulation of β1 and β2 integrin avidity by chemoattractants in eosinophils. Proc Natl Acad Sci USA 1996;93:10939–10944.
6.
Schleimer RP, Sterbinsky SA, Kaiser J, Bickel CA, Klunk DA, Tomioka K, Newman W, Luscinskas FW, Gimbrone MA Jr, McIntyre BW, Bochner BS: Interleukin-4 induces adherence of human eosinophils and basophils but not neutrophils to endothelium: Association with expression of VCAM-1. J Immunol 1992;148:1086–1092.
7.
Nagata M, Sedgwick JB, Bates ME, Kita H, Busse WW: Eosinophil adhesion to vascular cell adhesion molecule-1 activates superoxide anion generation. J Immunol 1995;155:2194–2202.
8.
Anwar ARF, Moqbel R, Walsh GM, Kay AB, Wardlaw AJ: Adhesion to fibronectin prolongs eosinophil survival. J Exp Med 1993;177:839–843.
9.
Anwar ARE, Walsh GM, Cromwell O, Kay AB, Wardlaw AJ: Adhesion to fibronectin primes eosinophil via α4/β1 (VLA-4). Immunology 1994;82:222–228.
10.
Horie S, Kita H: CD11b/CD18 (Mac-1) is required for degranulation of human eosinophils induced by human recombinant granulocyte-macrophage colony-stimulating factor and platelet-activating factor. J Immunol 1994;152:5457–5467.
11.
Hynes RO: Integrins: Versatility, modulation, and signaling in cell adhesion. Cell 1992;69:11–25.
12.
Matsumoto K, Sterbinsky SA, Bickel CA, Zhou DFH, Kovach NL, Bochner BS: Regulation of α4 integrin-mediated adhesion of human eosinophils to fibronectin and vascular cell adhesion molecule-1. J Allergy Clin Immunol 1997;99:648–656.
13.
Matsumoto K, Bochner BS, Wakiguchi H, Kurashige T: Altered expression of CD11b and CD62L after cross-linking of CD45 isoforms on human eosinophils. Int Arch Allergy Immunol 1998;117:S34–39.
14.
Wright MD, Tomlinson MG: The ins and outs of the transmembrane 4 superfamily. Immunol Today 1994;15:588–594.
15.
Rubinstein E, Le NF, Lagaudriere GC, Billard M, Conjeaud H, Boucheix C: CD9, CD63, CD81, and CD82 are components of a surface tetraspan network connected to HLA-DR and VLA integrins. Eur J Immunol 1996;26:2657–2665.
16.
Rubinstein E, Le NF, Billard M, Prenant M, Boucheix C: CD9 antigen is an accessory subunit of the VLA integrin complexes. Eur J Immunol 1994;24:3005–3013.
17.
Mannion BA, Berditchevski F, Kraeft SK, Chen LB, Hemler ME: Transmembrane-4 superfamily proteins CD81 (TAPA-1), CD82, CD63, and CD53 specifically associated with integrin α4β1 (CD49d/CD29). J Immunol 1996;157:2039–2047.
18.
Skubitz KM, Campbell KD, Iida J, Skubitz AP: CD63 associates with tyrosine kinase activity and CD11/CD18, and transmits an activation signal in neutrophils. J Immunol 1996;157:3617–3626.
19.
Maecker HT, Todd SC, Levy S: The tetraspanin superfamily: Molecular facilitators. FASEB J 1997;11:428–442.
20.
Kim JT, Gleich GJ, Kita H: Roles of CD9 molecules in survival and activation of human eosinophils. J Immunol 1997;159:926–933.
21.
Masellis SA, Shaw AR: CD9-regulated adhesion. Anti-CD9 monoclonal antibody induces pre-B cell adhesion to bone marrow fibroblasts through de novo recognition of fibronectin. J Immunol 1994;152:2768–2777.
22.
Masellis SA, Jensen GS, Seehafer JG, Slupsky JR, Shaw AR: Anti-CD9 monoclonal antibodies induce homotypic adhesion of pre-B cell lines by a novel mechanism. J Immunol 1990;144:1607–1613.
23.
Ebisawa M, Schleimer RP, Bickel CA, Bochner BS: Phenotyping of purified human peripheral blood eosinophils using the blind panel mAb; in Schlossman S, Boumsell L, Gilks W, Harlan J, Kishimoto T, Morimoto C, Ritz J, Shaw S, Silverstein R, Springer T, Tedder T, Todd R (eds): Leukocyte Typing V: White Cell Differentiation Antigens. New York, Oxford University Press, 1995, pp 1036–1038.
24.
Hansel TT, Pound JD, Pilling D, Kitas GD, Salmon M, Gentle TA, Lee SS, Thompson RA: Purification of human eosinophils by negative selection using immunomagnetic beads. J Immunol Methods 1989;122:97–103.
25.
Bochner BS, McKelvey AA, Schleimer RP, Hildreth JEK, MacGlashan DW Jr: Flow cytometric methods for analysis of human basophil surface antigens and viability. J Immunol Methods 1989;125:265–271.
26.
Matsumoto K, Schleimer RP, Saito H, Iikura Y, Bochner BS: Induction of apoptosis in human eosinophils by anti-Fas antibody treatment in vitro. Blood 1995;86:1437–1443.
27.
Peter MJ, Heyderman RS, Hatch DJ, Klein NJ: Investigation of platelet-neutrophil interactions in whole blood by flow cytometry. J Immunol Methods 1997;209:125–135.
28.
Bochner BS, Sterbinsky SA: Altered surface expression of CD11 and Leu 8 during human basophil degranulation. J Immunol 1991;146:2367–2373.
29.
Takahashi S, Doss C, Levy S, Levy R: TAPA-1, the target of an antiproliferative antibody, is associated on the cell surface with the Leu-13 antigen. J Immunol 1990;145:2207–2213.
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