Source: Annual Congress 2003 - Molecular biology of chronic lung disease and lung cancer
Disease area: Airway diseases

Abstract

Eosinophils are important in asthma pathophysiology, thus investigation of the mechanisms controlling their apoptosis is important. We examined membrane receptor expression, major basic protein (MBP) and eosinophil cationic protein (ECP) expression and apoptosis induction by mAb receptor ligation in the differentiated human eosinophilic line EOL-1. EOL-1 were differentiated by culturing with 0.1mM dibutyryl cyclic AMP for 9 days and 0.5mM butyric acid for 7 days. Membrane receptor expression by EOL-1 was assessed by immunostaining and flow cytometry. Expression of MBP and ECP were determined by immunohistochemistry. Apoptosis was measured by binding of FITC-annexin V to differentiated EOL-1 cultured for 20 hours alone or with CD45, CD45RA, CD45RB, CD45RO, FAS, CD69, and control mAb. EOL-1 expressed CD45, CD45RA, CD45RB, CD45RO, FAS, FAS-L, CD69, CD18 and VLA4. Positive immunostaining was observed in a proportion of differentiated EOL-1 for ECP (45% ± 5.5) and BMK13 (61% ± 4.5). EOL-1 cultured alone for 20 hours exhibited constitutive apoptosis (19% ± 3.3). mAb-treatment resulted in significant (P< 0.01) enhancement of EOL-1 constitutive apoptosis; i.e., CD45 (79% ± 6.6 [SEM]), CD45RA (65% ± 2.6), CD45RB (58% ± 5.1), FAS (42% ± 2.8), and CD69 (43% ± 4.1). CD45RO or control mAb had no significant effect on EOL-1 apoptosis induction. Differentiated EOL-1 exhibit mAb-induced apoptosis similar to that of peripheral blood eosinophils (Blaylock et al. J Allergy Clin Immunol 1999;104:1244). They may prove useful for studying the intracellular pathways controlling eosinophil apoptosis.

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M. W. Al-Rabia, M. G. Blaylock, D. W. Sexton, G. M. Walsh (Aberdeen, United Kingdom). Membrane receptor mediated apoptosis in the differentiated EOL-1 eosinophilic cell line. Eur Respir J 2003; 22: Suppl. 45, 1498

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