RESUMO

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.

Assuntos

Antígenos de Bactérias/imunologia , Antígenos de Bactérias/metabolismo , Lactuca/metabolismo , Vacina contra a Peste/imunologia , Vacina contra a Peste/metabolismo , Peste/imunologia , Proteínas Citotóxicas Formadoras de Poros/imunologia , Proteínas Citotóxicas Formadoras de Poros/metabolismo , Yersinia pestis/imunologia , Animais , Antígenos de Bactérias/genética , Western Blotting , Ensaio de Imunoadsorção Enzimática , Lactuca/genética , Camundongos , Camundongos Endogâmicos BALB C , Peste/microbiologia , Vacina contra a Peste/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Reação em Cadeia da Polimerase , Proteínas Citotóxicas Formadoras de Poros/genética , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/metabolismo , Rhizobium/genética , Yersinia pestis/patogenicidade