Expression of a modified Mannheimia haemolytica GS60 outer membrane lipoprotein in transgenic alfalfa for the development of an edible vaccine against bovine pneumonic pasteurellosis

J Biotechnol. 2008 Jun 1;135(2):224-31. doi: 10.1016/j.jbiotec.2008.03.006. Epub 2008 Mar 22.

Abstract

The GS60 antigen is one of the protective antigens of Mannheimia haemolytica A1. GS60 contains conserved domains belonging to the LppC family of bacterial outer membrane lipoproteins. A high antibody titer to GS60 has been shown to be significantly correlated with resistance to pneumonic pasteurellosis. Calves vaccinated with a commercial vaccine (Presponse) and demonstrating protection against M. haemolytica A1 produced antibodies directed against GS60. Alfalfa was chosen as the platform for an edible vaccine. Agrobacterium tumefaciens was used to mediate the transformation of alfalfa with sequences encoding a slightly shortened derivative of the GS60 antigen (GS60(54)). Stable transgenic alfalfa lines were recovered and production of GS60(54) was examined by Western immunoblot analysis. The antigen is stable in dried transgenic plant material stored at ambient temperature for more than a year. The plant-produced GS60(54) protein was shown to be immunogenic when injected into rabbits. Feeding of the dried transgenic alfalfa expressing the GS60(54) to rabbits is capable of inducing seroconversion, suggesting that GS60(54) could be an effective oral antigen for stimulating mucosal immune responses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood
  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Outer Membrane Proteins / immunology
  • Bacterial Outer Membrane Proteins / metabolism*
  • Bacterial Vaccines / genetics
  • Bacterial Vaccines / immunology*
  • Bacterial Vaccines / metabolism
  • Blotting, Western
  • Cattle
  • Cattle Diseases / immunology
  • Cattle Diseases / microbiology
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Mannheimia haemolytica / genetics
  • Mannheimia haemolytica / immunology
  • Mannheimia haemolytica / metabolism*
  • Medicago sativa / genetics
  • Medicago sativa / metabolism*
  • Microscopy, Fluorescence
  • Pasteurellosis, Pneumonic / blood
  • Pasteurellosis, Pneumonic / immunology*
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism
  • Rabbits
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Vaccination / methods
  • Vaccines, Edible / genetics
  • Vaccines, Edible / immunology
  • Vaccines, Edible / metabolism

Substances

  • Antibodies, Bacterial
  • Bacterial Outer Membrane Proteins
  • Bacterial Vaccines
  • Recombinant Fusion Proteins
  • Vaccines, Edible
  • Green Fluorescent Proteins