title: Evidence that Fc gamma receptors in rabbit yolk sac endoderm do not depend upon an acid pH to effect IgG binding and transcytosis in vitro.
abstract: An in vitro culture system has been devised creating apical and basal compartments separated by rabbit visceral yolk sac (VYS) with an intact epithelium. Selective transcytosis and binding of heterologous IgG applied to the apical yolk sac endoderm (YSE) was demonstrated in vitro using double label immunofluorescence. Thus, whilst both human and bovine IgG could be detected in endosomes in YSE, only human IgG could be detected in the basement membrane and vascular mesenchyme. This mirrors what is found in vivo. The Fc fragment of human Ig was transcytosed but not the Fab fragment, indicating that Fc receptors were expressed in the cultured YSE. When VYS was previously chilled to 4 degrees C to prevent endocytosis and treated with rabbit serum albumin to prevent non-specific binding, human IgG, but not bovine IgG, became specifically bound to YSE apical plasma membrane; comparison of binding at pH 6.0, 7.3 (the average pH of rabbit uterine fluid) and 8.0 revealed no obvious difference. Pre-exposure of VYS for up to 5 min in monensin, followed by culture in monensin and immunoglobulin-containing medium, did not prevent the selective transcystosis of human IgG, suggesting that an acidic compartment may not be needed for transcytosis. An acid pH dependent Fc gamma receptor equivalent to that on suckling rat gut jejunal enterocyte plasma membranes could not be isolated from rabbit YSE following exposure of solubilized membrane to affinity matrix bound IgG at pH 6.0 and elution at pH 8.0. These results contradict a recent suggestion that Fc receptors on all IgG transcytosing epithelia require an acid pH to effect IgG binding and selective transcytosis.
authors.affiliation: Department of Biology, School of Biological Sciences, University of Southampton, UK.
authors.forename: T J
authors.initials: TJ
authors.lastname: Meads
authors.forename: A E
authors.initials: AE
authors.lastname: Wild
mesh_terms: D000818:Animals; D002417:Cattle; D004705:Endocytosis; D004707:Endoderm / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000276:immunology / Q000378:metabolism*; D066298:In Vitro Techniques; D008875:Middle Aged; D008985:Monensin / Q000494:pharmacology; D011485:Protein Binding; D011817:Rabbits; D011961:Receptors, Fc / Q000378:metabolism*; D013696:Temperature; D015017:Yolk Sac / Q000378:metabolism* / Q000473:pathology
chemical_list: D007074:Immunoglobulin G; D011961:Receptors, Fc; D008985:Monensin