Transgenic mice secreting coronavirus neutralizing antibodies into the milk

J Virol. 1998 May;72(5):3762-72. doi: 10.1128/JVI.72.5.3762-3772.1998.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral
  • DNA, Complementary
  • Immunoglobulin A
  • Immunoglobulin Isotypes
  • Immunoglobulin alpha-Chains
  • Immunoglobulin kappa-Chains
  • Recombinant Proteins