Recombinant expression of polymeric IgA: incorporation of J chain and secretory component of human origin.
Mucosal J (joining) chain-expressing IgA immunocytes produce dimeric IgA that is actively transported by the epithelial polymeric Ig receptor (pIgR) to exocrine secretions. Release of secretory IgA (SIgA) occurs by cleavage of the covalently linked pIgR ectodomain, also known as bound secretory component. We have identified the human J-chain cDNA sequence through database screening, and isolated it from B cells for recombinant expression. Co-expression of this cDNA with an alpha heavy chain and a lambda light chain in Chinese hamster ovary (CHO) cells resulted in a mixture of recombinant monomeric and dimeric IgA in culture supernatants. This dimeric IgA was transported by the pIgR-mediated mechanism in vitro. Furthermore, expression of the human pIgR ectodomain together with the dimeric IgA, resulted in production of complete SIgA by the CHO cells. These results demonstrated that co-expression of the necessary polypeptide components allows a single mammalian cell to produce SIgA. Development of production systems for human antigen-specific recombinant SIgA may be important for applications in passive mucosal vaccination.
Johansen
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FE
Laboratory of Immunohistochemistry and Immunopathology (LIIPAT), Institute of Pathology, University of Oslo, Rikshospitalet, Norway. f.e.johansen@labmed.uio.no
Natvig Norderhaug
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I
Røe
M
M
Sandlie
I
I
Brandtzaeg
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D004285:Dogs; D015870:Gene Expression; D006801:Humans; D007070:Immunoglobulin A / Q000235:genetics* / Q000276:immunology; D007071:Immunoglobulin A, Secretory / Q000235:genetics* / Q000276:immunology; D007144:Immunoglobulin J-Chains / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology
D007070:Immunoglobulin A; D007071:Immunoglobulin A, Secretory; D007144:Immunoglobulin J-Chains; D011993:Recombinant Fusion Proteins; C034639:polymeric IgA