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

• Published in: Journal of biotechnology Vol. 135; no. 2; pp. 224 - 231
• Main Authors: Lee, Raymond W.H., Cornelisse, Mette, Ziauddin, Asma, Slack, Penelope J., Hodgins, Douglas C., Strommer, Judith N., Shewen, Patricia E., Lo, Reggie Y.C.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 01.06.2008; Amsterdam [New York, NY]: Elsevier; New York, NY Elsevier
• Subjects: Agrobacterium tumefaciens; alfalfa; Animals; Antibodies, Bacterial - blood; antibody formation; bacterial antigens; Bacterial Outer Membrane Proteins - genetics; Bacterial Outer Membrane Proteins - immunology; Bacterial Outer Membrane Proteins - metabolism; Bacterial Vaccines - genetics; Bacterial Vaccines - immunology; Bacterial Vaccines - metabolism; Biological and medical sciences; Biotechnology; Blotting, Western; Cattle; Cattle Diseases - immunology; Cattle Diseases - microbiology; Edible vaccine; edible vaccines; Fundamental and applied biological sciences. Psychology; gene expression; genetic transformation; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; GS60; lipoproteins; LppC; Mannheimia haemolytica; Mannheimia haemolytica - genetics; Mannheimia haemolytica - immunology; Mannheimia haemolytica - metabolism; Medicago sativa; Medicago sativa - genetics; Medicago sativa - metabolism; Microscopy, Fluorescence; outer membrane proteins; Pasteurellosis; Pasteurellosis, Pneumonic - blood; Pasteurellosis, Pneumonic - immunology; Plants, Genetically Modified - genetics; Plants, Genetically Modified - metabolism; Rabbits; recombinant antigens; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; seroconversion; transgenes; Transgenic alfalfa; transgenic plants; vaccination; Vaccination - methods; vaccine development; Vaccines, Edible - genetics; Vaccines, Edible - immunology; Vaccines, Edible - metabolism; Mannheimia haemolytica; Transgenic alfalfa; Edible vaccine; LppC; Pasteurellosis; GS60; Vertebrata; Mammalia; GS60;LppC;Pasteurellosis;Mannheimia haemolytica;Transgenic alfalfa;Edible vaccine; Bovine; Vaccine; Artiodactyla; Lipoprotein; External membrane; Transgenesis; Ungulata
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2008.03.006

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.