title: Production and characterization of recombinant IgA.
abstract: Existence of secretory immunity at the mucosal surfaces was first postulated in 1919. Since then experimental and clinical studies have indicated that it is immunoglobulin A (IgA) that provides the first line of immune defense at the mucosal surfaces. While a number of expression systems--including viral, plant and mammalian cells--have been used to produce recombinant IgA, we used the mammalian expression system to produce IgA1 and the three allotypes of IgA2. By introducing the gene coding for human secretory component (SC) into transfectants producing IgA1, we have generated a single mammalian cell system that produces covalently assembled secretory IgA (sIgA). Using pulse-chase analysis, we determined the covalent assembly pathways of IgA1, IgA2 and sIgA and identified some of the structural differences leading to the different assembly patterns. Using affinity purified proteins, we have shown that neither IgA1 nor any of the allotypes of IgA2 activate either the classical or the alternative complement pathways, but modulate the complement activity of IgG or IgM. The two N-linked glycosylation sites in IgA1 are not required for its binding to the polymeric Ig receptor (pIgR). Finally, we have shown that sIgA1 was more stable than dIgA1 in the gastrointestinal tract of mice, suggesting that SC provides resistance to IgA in the gastrointestinal tract.
authors.affiliation: Department of Microbiology and Molecular Genetics, Molecular Biology Institute, University of California, Los Angeles 90095, USA.
authors.forename: K R
authors.initials: KR
authors.lastname: Chintalacharuvu
authors.forename: S L
authors.initials: SL
authors.lastname: Morrison
mesh_terms: D000818:Animals; D002460:Cell Line; D005818:Genetic Engineering; D006801:Humans; D007071:Immunoglobulin A, Secretory / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D051379:Mice; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology
chemical_list: D007071:Immunoglobulin A, Secretory; D011994:Recombinant Proteins