CFTR localization in native airway cells and cell lines expressing wild-type or F508del-CFTR by a panel of different antibodies

被引:36
作者
Carvalho-Oliveira, I
Efthymiadou, A
Malhó, R
Nogueira, P
Tzetis, M
Kanavakis, E
Amaral, MD
Penque, D [1 ]
机构
[1] Inst Nacl Saude Dr Ricardo Jorge, Ctr Genet Humana, Observ Nacl Saude, P-1649016 Lisbon, Portugal
[2] Univ Athens, Aghia Sophia Childrens Hosp, Dept Med Genet, Athens, Greece
[3] Univ Lisbon, Dept Biol Vegetal, Fac Ciencias Lisboa, P-1699 Lisbon, Portugal
[4] Univ Lisbon, Dept Quim & Bioquim, Fac Ciencias Lisboa, P-1699 Lisbon, Portugal
关键词
cystic fibrosis; CFTR; CFTR antibody; immunocytochemistry; nasal brushing; epithelial cells;
D O I
10.1177/002215540405200207
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
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.
引用
收藏
页码:193 / 203
页数:11
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