Secretory component delays the conversion of secretory IgA into antigen-binding competent F(ab')2: a possible implication for mucosal defense.
Secretory component (SC) represents the soluble ectodomain of the polymeric Ig receptor, a membrane protein that transports mucosal Abs across epithelial cells. In the protease-rich environment of the intestine, SC is thought to stabilize the associated IgA by unestablished molecular mechanisms. To address this question, we reconstituted SC-IgA complexes in vitro by incubating dimeric IgA (IgAd) with either recombinant human SC (rSC) or SC isolated from human colostral milk (SCm). Both complexes exhibited an identical degree of covalency when exposed to redox agents, peptidyl disulfide isomerase, and temperature changes. In cross-competition experiments, 50% inhibition of binding to IgAd was achieved at approximately 10 nM SC competitor. Western blot analysis of IgAd digested with intestinal washes indicated that the alpha-chain in IgAd was primarily split into a 40-kDa species, a phenomenon delayed in rSC- or SCm-IgAd complexes. In the same assay, either of the SCs was resistant to degradation only if complexed with IgAd. In contrast, the kappa light chain was not digested at all, suggesting that the F(ab')2 region was left intact. Accordingly, IgAd and SC-IgAd digestion products retained functionality as indicated by Ag reactivity in ELISA. Size exclusion chromatography under native conditions of digested IgAd and rSC-IgAd demonstrates that SC exerts its protective role in secretory IgA by delaying cleavage in the hinge/Fc region of the alpha-chain, not by holding together degraded fragments. The function of integral secretory IgA and F(ab')2 is discussed in terms of mucosal immune defenses.
Crottet
P
P
Institut Suisse de Recherches Expérimentales sur le Cancer, Epalinges, Switzerland. blaise.corthesy@chuv.hospvd.ch
Corthésy
B
B
D000818:Animals; D000915:Antibody Affinity; D000942:Antigens, Bacterial / Q000378:metabolism; D001666:Binding Sites, Antibody; D001667:Binding, Competitive / Q000276:immunology; D019281:Dimerization; D010450:Endopeptidases / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis; D018928:Immunity, Mucosal; D007070:Immunoglobulin A / Q000378:metabolism; D007071:Immunoglobulin A, Secretory / Q000378:metabolism*; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000201:enzymology / Q000276:immunology*; D007700:Kinetics; D008070:Lipopolysaccharides / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008892:Milk / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D012635:Secretory Component / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
D000942:Antigens, Bacterial; D007070:Immunoglobulin A; D007071:Immunoglobulin A, Secretory; D007140:Immunoglobulin Fab Fragments; D008070:Lipopolysaccharides; D011994:Recombinant Proteins; D012635:Secretory Component; D010450:Endopeptidases