RESUMO
Transgenic plants represent an inexpensive alternative to classical fermentation systems for production of recombinant subunit vaccines. Transgenic potato plants were created that express the N-terminal domain of the glycoprotein S (N-gS) from Transmissible gastroenteritis coronavirus (TGEV), containing the major antigenic sites of the protein. Extracts from potato tubers expressing N-gS were inoculated intraperitoneally to mice, and the vaccinated mice developed serum IgG specific for TGEV. Furthermore, when potato tubers expressing N-gS were fed directly to mice, they developed serum antibodies specific for gS protein, demonstrating the oral immunogenicity of the plant derived spike protein from TGEV.
Assuntos
Coronavirus/imunologia , Glicoproteínas de Membrana/imunologia , Solanum tuberosum , Proteínas do Envelope Viral/imunologia , Administração Oral , Animais , Anticorpos Antivirais/sangue , Antígenos Virais/administração & dosagem , Coronavirus/química , Coronavirus/genética , Infecções por Coronavirus/prevenção & controle , Ensaio de Imunoadsorção Enzimática , Glicoproteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos BALB C , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Plasmídeos , Reação em Cadeia da Polimerase , Proteínas Recombinantes/administração & dosagem , Glicoproteína da Espícula de Coronavírus , Transformação Genética , Vacinas Sintéticas/administração & dosagem , Proteínas do Envelope Viral/genética , Vacinas Virais/administração & dosagemRESUMO
The major structural protein VP60 of rabbit hemorrhagic disease virus (RHDV) has been produced in transgenic potato plants under the control of a cauliflower mosaic virus 35S promoter or a modified 35S promoter that included two copies of a strong transcriptional enhancer. Both types of promoters allowed the production of specific mRNAs and detectable levels of recombinant VP60, which were higher for the constructs carrying the modified 35S promoter. Rabbits immunized with leaf extracts from plants carrying this modified 35S promoter showed high anti-VP60 antibody titers and were fully protected against the hemorrhagic disease.
Assuntos
Antígenos Virais/imunologia , Infecções por Caliciviridae/prevenção & controle , Capsídeo/imunologia , Vírus da Doença Hemorrágica de Coelhos/imunologia , Solanum tuberosum , Vacinas Sintéticas/imunologia , Proteínas Estruturais Virais/imunologia , Vacinas Virais/imunologia , Animais , Antígenos Virais/genética , Infecções por Caliciviridae/imunologia , Capsídeo/genética , Vírus da Doença Hemorrágica de Coelhos/genética , Plantas Geneticamente Modificadas , Coelhos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Vacinas Sintéticas/genética , Proteínas Estruturais Virais/genética , Vacinas Virais/genéticaRESUMO
Sequences encoding the immunoglobulin heavy-chain variable (VH) domains were engineered in a new general purpose vector to transform plants via Agrobacterium. The expression of an isolated VH domain (IVD) after introduction into the plant genome has been monitored by northern, western and immunohistochemical analysis. Immunoblotting showed that the polypeptide was stably expressed and accounted for up to 1% of the soluble protein fraction. It is therefore proposed that single immunoglobulin domains of suitable specificity expressed in plants may constitute an effective system to inhibit the activity of molecules involved in plant pathology or plant development.