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Title: IMMUNOGENICITY OF A NOVEL, BIVALENT, PLANT-BASED ORAL VACCINE AGAINST HEPATITIS B AND HUMAN IMMUNODEFICIENCY VIRUSES

Author
item SHCHELKUNOV, SERGEI - KOLTSOVO RUSSIA
item SALYAEV, RURIK - IRKUTSK RUSSIA
item POZDNYAKOV, SERGEI - KOLTSOVO RUSSIA
item REKOSLAVSKAYA, NATALIA - IRKUTSK RUSSIA
item NESTEROV, ANDREI - KOLTSOVO RUSSIA
item RYZHOVA, TATIANA - NOVOSIBIRSK RUSSIA
item SUMTSOVA, VALENTINA - IRKUTSK RUSSIA
item PAKOVA, NATALIA - IRKUTSK RUSSIA
item MISHUTINA, ULIANA - IRKUTSK RUSSIA
item KOPYTINA, TATIANA - IRKUTSK RUSSIA
item Hammond, Rosemarie

Submitted to: Biotechnology Letters
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/1/2006
Publication Date: 3/1/2006
Citation: Shchelkunov, S.N., Salyaev, R.K., Pozdnyakov, S.G., Rekoslavskaya, N.I., Nesterov, A.E., Ryzhova, T.S., Sumtsova, V.M., Pakova, N.V., Mishutina, U.O., Kopytina, T.V., Hammond, R. 2006. Immunogenicity of a novel, bivalent, plant-based oral vaccine against hepatitis b and human immunodeficiency viruses. Biotechnology Letters. 28:959-967.

Interpretive Summary: In recent years, plants have been used successfully for the large-scale production of bioengineered vaccines, antibodies, and animal proteins, such as growth hormone or blood substitutes, for medical and veterinary use. We produced an artificial protein as a candidate vaccine against Human immunodeficiency virus (HIV) and hepatitis B virus (HBV) in genetically engineered tomatoes. Oral administration of the candidate vaccine, present in dried tomato fruit, to experimental mice resulted in an immune response to the artificial protein. This result suggests that ingestion of the genetically engineered vaccine can be used effectively to stimulate potentially protective antibody responses in humans. This plant-produced protein has the potential for development into a candidate vaccine for vaccination of humans against HIV and HBV. The report will be of interest to an international audience of researchers, clinicians, and representatives of industry, academia, and government organizations with an interest in plant-based systems for production of recombinant medicines, vaccinology, immunology, and human and animal health.

Technical Abstract: The synthetic chimeric gene TBI-HBS, encoding the immunogenic ENV and GAG epitopes of Human immunodeficiency virus (HIV)-1 (immunogens of T- and B-lymphocytes) and the surface protein (HBsAg) of the hepatitis B virus (HBV), was introduced into tomato plants var. Ventura by means of the agrobacterial vector pBINp35STBI-HBS, and transgenic tomato plants with the integrated gene TBI-HBS were generated. The synthesis of chimeric TBI-HBS protein in fruits of transgenic tomato plants was verified by immunoassays. The fruits of transgenic tomato of the T1 generation were fed to experimental mice for a period of one week. On the 14th and 28th days post-feeding, high levels of antibodies specific to the antigenic proteins of HIV and HBV were found in the serum of the test animals. HIV- and HBV-specific antibodies were also found in their feces. There were no HIV- or HBV-specific antibodies in the control group of mice. Intraperitoneal injection of a DNA vaccine directing synthesis of the same TBI-HBsAg protein boosted the antibody response to HIV in the blood serum, however, had it no effect on the high level of antibodies produced to HBV. In summary it was found that the antigens HBsAg (HBV) and TBI (HIV-1) were synthesized in fruits of transgenic tomato as a result of the integration of the construct pBINp35STBI-HBS and that expression of the chimeric protein TBI-HBS was at a level that was able to induce the formation of an immunogenic response in mice after oral delivery of the vaccine.