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Published ahead of print on June 30, 2005, doi:10.1165/rcmb.2005-0132OC
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American Journal of Respiratory Cell and Molecular Biology. Vol. 33, pp. 363-370, 2005
© 2005 American Thoracic Society
DOI: 10.1165/rcmb.2005-0132OC

Antibodies to the Polymeric Immunoglobulin Receptor with Different Binding and Trafficking Patterns

Sanhita Gupta, Michael Heacock, Aura Perez and Pamela B. Davis

Department of Pediatrics, Case Western Reserve University School of Medicine, Cleveland, Ohio

Correspondence and requests for reprints should be addressed to Pamela B. Davis, Department of Pediatrics, Case Western Reserve University at Rainbow Babies and Childrens Hospital, Biomedical Research Bldg, Rm 831, 2109 Adelbert Road, Cleveland, OH 44106-6006. E-mail: pbd{at}po.cwru.edu

The polymeric immunoglobulin receptor (pIgR) has been proposed as a therapeutic target, but its potential depends on the efficiency of uptake and trafficking of the receptor ligand. Mouse monoclonal antibodies (Mabs) directed against pIgR, selected for strong binding to secretory component (SC) and secretory IgA (sIgA), were tested in a transcytosis assay in 16HBEo– cells (human bronchial epithelial cell line) transfected with human pIgR. Intracellular trafficking was followed by confocal microscopy. Mabs fell into two classes. For two Mabs, transcytosis from basolateral to apical surface is rapid, unidirectional, and little Mab is retained in the cell. For three Mabs, basolateral to apical transcytosis occurs to a significantly lesser extent, reverse transcytosis is permitted, and some of the Mab is retained in the perinuclear region even after 24 h. When tested for their ability to recognize and immunoprecipitate pIgR with systematic truncations and deletions of the five immunoglobulin (Ig)-like domains, all Mabs bound to the fifth Ig-like domain, but three of them also bound to the C-terminal region of pIgR near the plasma membrane. Different binding sites probably account for the different trafficking of these Mabs and may predict differential therapeutic utility.

Key Words: epithelial cells, basolateral and apical • pIgR • transcytosis







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Proc. Am. Thorac. Soc. Am. J. Respir. Crit. Care Med.
Copyright © 2005 American Thoracic Society.