Nonspecific binding of common anti-CFTR antibodies in ciliated cells of human airway epithelium View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2021-12-01

AUTHORS

Yukiko Sato, Kamila R. Mustafina, Yishan Luo, Carolina Martini, David Y. Thomas, Paul W. Wiseman, John W. Hanrahan

ABSTRACT

There is evidence that the cystic fibrosis transmembrane conductance regulator (CFTR) anion channel is highly expressed at the apical pole of ciliated cells in human bronchial epithelium (HBE), however recent studies have detected little CFTR mRNA in those cells. To understand this discrepancy we immunostained well differentiated primary HBE cells using CFTR antibodies. We confirmed apical immunofluorescence in ciliated cells and quantified the covariance of the fluorescence signals and that of an antibody against the ciliary marker centrin-2 using image cross-correlation spectroscopy (ICCS). Super-resolution stimulated emission depletion (STED) imaging localized the immunofluorescence in distinct clusters at the bases of the cilia. However, similar apical fluorescence was observed when the monoclonal CFTR antibodies 596, 528 and 769 were used to immunostain ciliated cells expressing F508del-CFTR, or cells lacking CFTR due to a Class I mutation. A BLAST search using the CFTR epitope identified a similar amino acid sequence in the ciliary protein rootletin X1. Its expression level correlated with the intensity of immunostaining by CFTR antibodies and it was detected by 596 antibody after transfection into CFBE cells. These results may explain the high apparent expression of CFTR in ciliated cells and reports of anomalous apical immunofluorescence in well differentiated cells that express F508del-CFTR. More... »

PAGES

23256

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/s41598-021-02420-x

DOI

http://dx.doi.org/10.1038/s41598-021-02420-x

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https://app.dimensions.ai/details/publication/pub.1143524012

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/34853321


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