Production of human rotavirus and Salmonella antigens in plants and elicitation of fljB-specific humoral responses in mice.
A Nicotiana benthamiana transient expression system was used to express single antigen and dimeric combinations of the human rotavirus (HRV) VP7 and a truncated VP4 (VP4Δ) proteins fused with Salmonella typhimurium's flagellin fljB subunit. Immunoblot analyses using rabbit antibodies generated against these proteins demonstrated that the constructs were successfully expressed with yields ranging from 0.85 to 31.97 μg of recombinant protein per gram of fresh leaf tissue. Expressing the single and dimeric antigens has no effect on plant growth and development except for VP7 and VP4Δ::VP7, which show mild necrotic lesions. Immunization of mice with proteins from leaves transformed with constructs bearing the fljB moiety elicited an fljB-specific humoral response. The Nicotiana benthamiana transient system is efficient to express multiple combinations of pathogen proteins and demonstrates the potential of generating a Salmonella typhimurium subunit vaccine in plants.
Bergeron-Sandoval
Louis-Philippe
LP
Département des Sciences Biologiques, Université du Québec à Montréal, CP 8888 Succ. Centre-ville, Montreal, QC H3C 3P8, Canada.
Girard
Aurélie
A
Ouellet
François
F
Archambault
Denis
D
Sarhan
Fathey
F
D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D000956:Antigens, Viral / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D002478:Cells, Cultured; D005260:Female; D005408:Flagellin / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D006801:Humans; D056724:Immunity, Humoral / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012401:Rotavirus / Q000235:genetics / Q000276:immunology*; D012475:Salmonella / Q000235:genetics / Q000276:immunology*; D014026:Nicotiana / Q000235:genetics / Q000276:immunology*
D000942:Antigens, Bacterial; D000956:Antigens, Viral; D036022:Capsid Proteins; C094427:FljB protein, Salmonella; D011993:Recombinant Fusion Proteins; C060339:VP4 protein, Rotavirus; C050779:VP7 protein, Rotavirus; D005408:Flagellin