title: Expression of biologically active human interferon alpha 2 in Aloe vera.
abstract: Methods necessary for the successful transformation and regeneration of Aloe vera were developed and used to express the human protein, interferon alpha 2 (IFNα2). IFNα2 is a secreted cytokine that plays a vital role in regulating the cellular response to viral infection. Transgenic plants were regenerated from callus cultures initiated from zygotic embryos. Expression of the IFNA2 transgene in transformed plants was confirmed by RT-PCR and IFNα2 protein was detected by immunoblot analysis. Human A549 cells treated with transgenic aloe extracts for 6 h induced expression of the interferon stimulated gene 54, indicating activation of the IFN signaling pathway. The biological activity of the aloe produced IFNα2 was assessed using an antiviral assay with A549 cells treated with extracts from both the rind and pulp fractions of the shoot and subsequently infected with the lytic encephalomyocarditis virus. The highest level of activity attributable to recombinant IFNα2 was determined to be 625 IU/mg of total soluble protein (TSP) in the rind and 2,108 IU/mg TSP in the pulp. Two daughter plants that vegetatively budded during the course of this study were also confirmed to express IFNα2. These results confirm that Aloe vera is capable of expressing a human protein with biological activity, and that a secreted protein targeting the apoplast can be detected in the pulp fraction of the plant.
authors.affiliation: Thegreencell, Inc., 505 Capitol Court NE, Suite 200, Washington, DC 20002, USA. williaml@thegreencell.com
authors.forename: William
authors.initials: W
authors.lastname: Lowther
authors.forename: Kevin
authors.initials: K
authors.lastname: Lorick
authors.forename: Susan D
authors.initials: SD
authors.lastname: Lawrence
authors.forename: Wen-Shuz
authors.initials: WS
authors.lastname: Yeow
mesh_terms: D000504:Aloe / Q000737:chemistry* / Q000235:genetics; D000998:Antiviral Agents / Q000494:pharmacology*; D004686:Encephalomyocarditis virus / Q000187:drug effects*; D018745:Genome, Plant; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D016898:Interferon-alpha / Q000235:genetics / Q000378:metabolism*; D010936:Plant Extracts / Q000494:pharmacology*; D018515:Plant Leaves / Q000187:drug effects; D030821:Plants, Genetically Modified / Q000235:genetics*; D012639:Seeds / Q000737:chemistry*; D015398:Signal Transduction / Q000187:drug effects; D019076:Transgenes / Q000502:physiology
chemical_list: D000998:Antiviral Agents; C554158:IFNA2 protein, human; D016898:Interferon-alpha; D010936:Plant Extracts; D005966:Glucuronidase