2B4 (CD244) is Involved in Eosinophil Adhesion and Chemotaxis,and its Surface Expression is Increased in Allergic Rhinitis after Challenge

Abstract

A role for the subtypes of CD2 Ig superfamily receptors has been recently demonstrated in eosinophilic inflammation in experimental asthma and atopic asthmatics. We investigated the functions of 2B4 (CD244) molecules in eosinophil adhesion and chemotaxis, and correlated the results to the pathophysiology of allergic rhinitis (AR). Herein, we show that agonistic stimulation of 2B4 by C1.7, the anti-human 2B4 functional grade purified antibody, resulted in significant increase of eosinophils and eosinophil cell line (Eol-1 cells) adhesion to collagen type IV, and random migration. These functions were associated with tyrosine kinase phosphorylation of several protein residues of low molecular weight. Flow cytometry (FACS) experiments demonstrated that Eol-1 cells, normal peripheral blood eosinophils and eosinophils from AR patients, express surface 2B4 molecules. In vitro AR model demonstrated that the CC-chemokine receptor CCR3 stimulation by eotaxin induced significant increase in the expression of surface 2B4 in eosinophils and Eol-1 cells. Immunofluorescence confocal microscopy images showed that eotaxin induces also redistribution of 2B4 molecules towards the pseudopods in eosinophils and Eol-1 cells, changing their shape. Blocking of 2B4 molecules by the corresponding neutralizing antibody inhibited eotaxin induced Eol-1-adhesion, chemotaxis and the cytoskeleton changes. Pretreatment of Eol-1 cells with 1 μM genistein blocked eotaxin-induced Eol-1 adhesion, chemotaxis and 2B4 up-regulated expression. In vivo correlation demonstrated the expression of 2B4 molecules in eosinophils from AR patients to be significantly increased, after nasal provocation challenge. These results identify a novel role for 2B4 molecules in eosinophil functional migratory response and may point to a novel tyrosine kinase-mediated ligation between CCR3 receptor and 2B4 co-receptor in eosinophil chemotaxis. If so, then 2B4 molecules would be a novel target for therapeutic modalities in diseases characterized by eosinophilia such as AR.

Keywords

eosinophils Eol-I chemotaxis adhesion allergic rhinitis 2B4 (CD244)CD2 tyrosine kinase

References

Coyle

Gutierrez-Ramos

. The expanding B7 superfamily, increasing complexity in costimulatorty signals regulating T cell function. Nat Immunol 2001; 2:203–209.

Carreno

Collins

. The B7 family of ligands and its receptors: New pathways for costimulation and inhibition of immune responses. Ann Rev Immunol 2002; 20:29–53.

Quezada

Jarvinen

Lind

. CD40/CD154 interactions at interface of tolerance and immunity. Ann Rev Immunol 2004; 22:307–28.

Engel

Eck

Terhorst

. The SAP and SLAM families in immune responses and X-linked lymphoproliferative disease. Nat Rev Immunol 2003; 3:813–21.

Mathew

Garni-Wagner

Land

. Cloning and characterization of the 2B4 gene encoding a molecule associated with non-MHC-restricted killing mediated by activated natural killer cells and T cells. J Immunol 1993; 151:5328–37.

Valiante

Trinchieri

. Identification of a novel signal transduction surface molecule on human cytotoxic lymphocytes. J Exp Med 1993; 178:1397–406.

Veillette

Latour

. The SLAM family of immune-cell receptors. Curr Opin Immunol 2003; 15:277–85.

Nakajima

Cella

Langen

. Activating interactions in human NK cell recognition: The role of 2B4-CD48. Eur J Immunol 1999; 29:1676–83.

Boles

Nakajima

Colonna

. Molecular characterization of a novel human natural cell receptor homologous to mouse 2B4. Tissue Antigens 1999; 54:27–34.

10.

Romero

Benitez

March

. Differential expression of SAP and EAT-2 binding leukocyte cell-surface molecules CD48, CD 150 (SLAM), CD229 (Ly9) and CD244 (2B4). Tissue Antigens 2004; 64:132–44.

11.

Latchman

McKay

Reiser

. Identification of the 2B4 molecule as a counter-receptor for CD48. J Immunol 1998; 161:5809–12.

12.

Brown

Boles

van der Merwe

. 2B4, the natural killer and T cell immunoglobulin superfamily surface protein, is a ligand for CD48. J Exp Med 1998; 188:2083–90.

13.

Munitz

Bachelet

Eliashar

. CD48 is an allergen and IL-13-induced activation molecule on eosinophils. J Immunol 2006; 177:77–83.

14.

Munitz

Bachelet

Finkelman

. CD48 is critically involved in allergic eosinophilic airway inflammation. Am J Respir Crit Care Med 2007; 175:911–18.

15.

Munitz

Bachelet

Fraenkel

. 2B4 (CD244) is expressed and functional on human eosinophils. J Immunol 2005; 174:110–18.

16.

Weller

. Eosinophilia. J Allergy Clin Immunol 1984;73:1–14.

17.

Ponath

Qin

Ringler

. Cloning of the human eosinophil chemoattractant, eotaxin expression, receptor binding, and functional properties suggest a mechanism for the selective recruitment of eosinophils. J Clin Invest 1996; 97:604–12.

18.

Ponath

Qin

Post

. Molecular cloning and characterization of a human eotaxin receptor expressed selectively on eosinophils. J Exp Med 1996; 183:2437–48.

19.

Gracia-Zepeda

Rothenberg

Ownbey

. Human eotaxin is a specific chemoattractant for eosinophil cells and provides a new mechanism to explain tissue eosinophilia. Nature Med 1996; 2:449–56.

20.

Lalaker

Nkrumah

Lee

. Chitin stimulates expression of acidic mammalian chitinase and eotaxin-3 by human sinonasal epithelial cells in vitro. Am J Rhinol Allergy 2009; 23:8–14.

21.

Sun

Kantarjian

. Protein expression profiling of cytokines and cytokine receptors on purified chronic lymphocytic leukemia cells from patients with favourable prognostic indicators. Leuk Lymphoma 2008; 49:751–56.

22.

Yoshida

Aizu-Yokota

Sonoda

. Production and regulation of eotaxin-2/CCL24 in a differentiated human leukemic cell line, HT93. Biol Pharm Bull 2007; 30:1826–32.

23.

Ishihara

Takahashi

Kaneko

. Differentiation of eosinophilic leukemia Eol-1 cells into eosinophils induced by histone deacetylase inhibitors. Life Sciences 2001; 80:1213–20.

24.

El-Shazly

Berger

Girodet

. Fraktalkine produced by airway smooth muscle cells contributes to mast cell recruitment in asthma. J Immunol 2006; 176:1860–68.

25.

El-Shazly

Yamaguchi

Masuyama

. Novel association of the Src family kinases, Hck and c-Fgr, with CCR3 receptor stimulation: A possible mechanism for eotaxin-induced human eosinophil chemotaxis. Bioch Biophys Res Commun 1999; 264:163–70.

26.

El-Shazly

Keisuke

Eura

. Mechanisms involved in human eosinophil chemotaxis induced by the newly cloned C-C chemokine eotaxin. Allergol Int 1998; 47:49–12.

27.

Campo

PMH

Almeida

Sanchez

. Normal patterns of expression of glycosylphosphatidyl linositol-anchored proteins on different subsets of peripheral blood cells: A frame of reference for the diagnosis of paroxysmal nocturnal haemoglobinuria. Cytometry part B (Clinical cytometry) 2006; 70B: 71–81.

28.

Ridley

Schwartz

Burridge

. Cell migration: Integrating signals from front to back. Science 2003; 302:1704–09.

29.

Kiema

Lad

Jiang

Oxley

. The molecular basis of filamin binding to integrins and competition with talin. Mol Cell 2006; 21:337–47.

30.

Chen

Relouzat

Roncagalli

. Molecular dissection of 2B4 signaling: Implications for signal transduction by SLAM-related receptors. Mol Cell Biol 2004; 24:5144–56.

31.

Eissmann

Beauchamp

Wooters

. Molecular basis for positive and negative signaling by the natural killer cell receptor 2B4 (CD244). Blood 2005; 105:4722–29.