Expression of an immunogenic F1-V fusion protein in lettuce as a plant-based vaccine against plague

• Published in: Planta 2010-07, Vol.232 (2), p.409-416
• Main Authors: Rosales-Mendoza, Sergio, Soria-Guerra, Ruth E, Moreno-Fierros, Leticia, Alpuche-Solís, Ángel G, Martínez-González, Luzmila, Korban, Schuyler S
• Format: Article
• Language: English
• Subjects: Agriculture; Animals; Antibodies; Antigenic fusion protein; Antigens; Antigens, Bacterial - genetics; Antigens, Bacterial - immunology; Antigens, Bacterial - metabolism; Biological and medical sciences; Biomedical and Life Sciences; Blotting, Western; Ecology; Enzyme-Linked Immunosorbent Assay; Forestry; Fundamental and applied biological sciences. Psychology; Immunization; Lactuca - genetics; Lactuca - metabolism; Lettuce; Life Sciences; Mice; Mice, Inbred BALB C; Original Article; Plague; Plague - immunology; Plague - microbiology; Plague Vaccine - genetics; Plague Vaccine - immunology; Plague Vaccine - metabolism; Plant antigens; Plant Sciences; Plant-based vaccine; Plants; Plants, Genetically Modified - genetics; Plants, Genetically Modified - metabolism; Polymerase Chain Reaction; Pore Forming Cytotoxic Proteins - genetics; Pore Forming Cytotoxic Proteins - immunology; Pore Forming Cytotoxic Proteins - metabolism; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - immunology; Recombinant Fusion Proteins - metabolism; Rhizobium - genetics; Transgenes; Transgenic plants; Vaccination; Vaccines; Vegetables; Yersinia pestis; Yersinia pestis - immunology; Yersinia pestis - pathogenicity
• ISSN: 0032-0935; 1432-2048
• DOI: 10.1007/s00425-010-1176-z

Yersinia pestis is a pathogenic agent that causes the bubonic and pneumonic plague. The development of an efficient and low-cost oral vaccine against these diseases is highly desirable. In this study, the immunogenic fusion protein F1-V from Y. pestis was introduced into lettuce via Agrobacterium-mediated transformation, and putative transgenic lines were developed. The presence of the transgene in these putative transgenic lines was determined using polymerase chain reaction (PCR), and transgene integration and transgene copy number were confirmed following Southern blot analysis. The presence of specific F1-V transcripts was confirmed by reverse-transcriptase (RT)-PCR. Using monoclonal antibodies, ELISA and western blot analysis revealed that the expected antigenic F1-V protein was successfully expressed in transgenic lines. Mice immunized subcutaneously with lettuce expressing the F1-V antigen developed systemic humoral responses as ‘proof of concept' of using lettuce as a production platform for the F1-V immunogen that could be used as a candidate plant-based vaccine against plague.