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Transgenic mice secreting coronavirus neutralizing antibodies into the milk

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  • I Sola
  • J Castilla
  • B Pintado
  • J M Sánchez-Morgado
  • C B Whitelaw
  • A J Clark
  • L Enjuanes

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    Rights statement: Copyright © 1998, American Society for Microbiology

    Final published version, 452 KB, PDF-document

Original languageEnglish
Pages (from-to)3762-72
Number of pages11
JournalJournal of Virology
Volume72
Issue number5
Publication statusPublished - May 1998

Abstract

Ten lines of transgenic mice secreting transmissible gastroenteritis coronavirus (TGEV) neutralizing recombinant monoclonal antibodies (rMAbs) into the milk were generated. The rMAb light- and heavy-chain genes were assembled by fusing the genes encoding the variable modules of the murine MAb 6A.C3, which binds an interspecies conserved coronavirus epitope essential for virus infectivity, and a constant module from a porcine myeloma with the immunoglobulin A (IgA) isotype. The chimeric antibody led to dimer formation in the presence of J chain. The neutralization specific activity of the recombinant antibody produced in transiently or stably transformed cells was 50-fold higher than that of a monomeric rMAb with the IgG1 isotype and an identical binding site. This rMAb had titers of up to 10(4) by radioimmunoassay (RIA) and neutralized virus infectivity up to 10(4)-fold. Of 23 transgenic mice, 17 integrated both light and heavy chains, and at least 10 of them transmitted both genes to the progeny, leading to 100% of animals secreting functional TGEV neutralizing antibody during lactation. Selected mice produced milk with TGEV-specific antibody titers higher than 10(6) as determined by RIA, neutralized virus infectivity by 10(6)-fold, and produced up to 6 mg of antibody per ml. Antibody expression levels were transgene copy number independent and integration site dependent. Comicroinjection of the genomic beta-lactoglobulin gene with rMAb light- and heavy-chain genes led to the generation of transgenic mice carrying the three transgenes. The highest antibody titers were produced by transgenic mice that had integrated the antibody and beta-lactoglobulin genes, although the number of transgenic animals generated does not allow a definitive conclusion on the enhancing effect of beta-lactoglobulin cointegration. This approach may lead to the generation of transgenic animals providing lactogenic immunity to their progeny against enteric pathogens.

    Research areas

  • Amino Acid Sequence, Animals, Antibodies, Monoclonal, Antibodies, Viral, Base Sequence, COS Cells, Cell Line, Cell Line, Transformed, DNA, Complementary, Humans, Immunoglobulin A, Immunoglobulin Isotypes, Immunoglobulin alpha-Chains, Immunoglobulin kappa-Chains, Mice, Mice, Transgenic, Milk, Molecular Sequence Data, Neutralization Tests, Recombinant Proteins, Swine, Transmissible gastroenteritis virus, Tumor Cells, Cultured

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