Antibodies to the polymeric immunoglobulin receptor with different binding and trafficking patterns.
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.
Department of Pediatrics, Case Western Reserve University School of Medicine, Cleveland, OH, USA.
Sanhita
S
Gupta
Michael
M
Heacock
Aura
A
Perez
Pamela B
PB
Davis
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D001692:Biological Transport / Q000502:physiology; D016764:Cell Polarity; D002469:Cell Separation; D002478:Cells, Cultured; D005434:Flow Cytometry; D006801:Humans; D007071:Immunoglobulin A, Secretory / Q000378:metabolism; D051379:Mice; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D019056:Receptors, Polymeric Immunoglobulin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012635:Secretory Component / Q000378:metabolism
D000911:Antibodies, Monoclonal; D007071:Immunoglobulin A, Secretory; D019056:Receptors, Polymeric Immunoglobulin; D012635:Secretory Component