| Abstract Detail
Plant biotech & Risk Assessment Wang, David J [1], Ma, Shengwu [2]. Production of Biologically Active Human IL-13 in Transgenic Plants for Oral Immunotherapy against Type 1 Diabetes. Interleukin-13 (IL-13) is a pleiotropic regulatory cytokine that plays a protective role against many immune-associated diseases including type 1 diabetes. However, the high production cost of recombinant IL-13 (rIL-13) through conventional expression methods has limited broader clinical testing, especially for the more convenient and safer oral delivery of IL-13 as opposed to parental injection in patients. Here, we report the use of transgenic plants as an alternative expression system for the cost-effective production of recombinant human IL-13 (rhIL-13). A DNA construct containing a full-length cDNA of the human IL-13 under the transcriptional control of two tandem copies of the strong constitutive promoter cauliflower mosaic virus 35S (CaMV35S) was produced and introduced into tobacco by Agrobacterium-mediated transformation. Transcriptional expression of the introduced hIL-13 gene in transgenic plants was confirmed by RT-PCR and northern blot. Immunoblotting analysis coupled with enzymatic deglycosylation demonstrated that the human IL-13 protein is expressed as three major glycosylated peptides. The expression levels of rhIL-13 in tobacco leaf tissues reach as high as 0.15% of total soluble protein (TSP). In vitro bioassays using a factor-dependent human erythroleukemic cell line, TF-1, showed that plant-derived rhIL-13 retained the biological functions of the authentic hIL-13 protein. Furthermore, the plant-produced IL-13 was found to be more resistant to pancreatic proteolysis. More importantly, oral administration of IL-13 transgenic plant tissues reduced the incidence of diabetes in NOD mice. These results suggest that transgenic plants may not only provide a new source of inexpensive human IL-13, but may also be used as a convenient vehicle for effective oral delivery of hIL-13 to prevent and treat clinical diseases such as type 1 diabetes. Log in to add this item to your schedule
1 - A.B. Lucas Secondary School, Science, 656 Tennet Ave, London, Ontario, N5X 1L8, Canada 2 - University of Western Ontario, Department of Biology, Biological & Geological Building, 1151 Richmond St, London, Ontario, N6A 5B7, Canada
Keywords: IL-13 Type 1 diabetes transgenic.
Presentation Type: Plant Biology Abstract Session: P Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton Date: Sunday, July 8th, 2007 Time: 8:00 AM Number: P45004 Abstract ID:258 |