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Journal of Histochemistry and Cytochemistry
Volume 52 (2): 193-203, 2004
Copyright ©The Histochemical Society, Inc.

CFTR Localization in Native Airway Cells and Cell Lines Expressing Wild-type or F508del-CFTR by a Panel of Different Antibodies

Isabel Carvalho–Oliveira, Alexandra Efthymiadou, Rui Malhó, Paulo Nogueira, Maria Tzetis, Emmanuel Kanavakis, Margarida D. Amaral and Deborah Penque

Centro de Genética Humana (ICO,MDA,DP), Observatório Nacional de Saúde (PN), Instituto Nacional de Saúde Dr Ricardo Jorge, Lisboa, Portugal; Department of Medical Genetics, University of Athens, Aghia Sophia Children's Hospital, Athens, Greece (AE,MT,EK); and Departamento de Biologia Vegetal (RM), Departamento de Química e Bioquímica (MDA), Faculdade de Ciências de Lisboa, Universidade de Lisboa, Lisboa, Portugal.

Correspondence to: Deborah Penque, PhD, Centro de Genética Humana, Instituto Nacional de Saúde Dr Ricardo Jorge, 1649-016 Lisboa, Portugal. E-mail: deborah.penque{at}insa.min-saude.pt

The intracellular localization of cystic fibrosis transmembrane conductance regulator (CFTR) in native tissues is a major issue in the study of mutation, processing, and trafficking effects in CFTR and in the evaluation of therapeutic strategies in cystic fibrosis (CF). This work evaluated the applicability of ten different antibodies (Abs) under various fixation techniques for CFTR localization in fresh-brushed nasal epithelial cells collected from CF patients homozygous for F508del and control individuals. In parallel, the same Ab panel was also tested on BHK cell lines overexpressing wild-type or F508del CFTR. The Abs MATG1061, 169, Lis1, MP-CT1, CC24-R, MAB25031, and MAB1660 gave the best detection of CFTR in the apical region (AR) of nasal tall columnar epithelial (TCE) cells. The labeling pattern of these Abs was consistent with the postulated processing defect of F508del CFTR because only a minority of CF TCE cells present CFTR in the AR. In contrast, M3A7, MM13-4, and L12B4 weakly react with the AR and stain almost exclusively a cis-Golgi-like structure in the majority of CF and non-CF airway cells. In BHK cells, all the Abs enabled distinction between wild-type CFTR localization in cell membrane from F508del CFTR, which in these cells is exclusively located in the endoplasmic reticulum. (J Histochem Cytochem 52:193–203, 2004)

Key Words: cystic fibrosis • CFTR • CFTR antibody • immunocytochemistry • nasal brushing • epithelial cells


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