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  Viral and murine interleukin-10 are correctly processed and retain their biological activity when produced in tobacco

Bortesi, L., Rossato, M., Schuster, F., Raven, N., Stadlmann, J., Avesani, L., et al. (2009). Viral and murine interleukin-10 are correctly processed and retain their biological activity when produced in tobacco. BMC Biotechnology, 9, 22. doi:10.1186/1472-6750-9-22.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0014-2619-9 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0014-261A-7
Genre: Journal Article

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 Creators:
Bortesi, L.1, Author
Rossato, M.1, Author
Schuster, F.1, Author
Raven, N.1, Author
Stadlmann, J.1, Author
Avesani, L.1, Author
Falorni, A.1, Author
Bazzoni, F.1, Author
Bock, R.2, Author              
Schillberg, S.1, Author
Pezzotti, M.1, Author
Affiliations:
1External Organizations, ou_persistent22              
2Organelle Biology and Biotechnology, Department Bock, Max Planck Institute of Molecular Plant Physiology, Max Planck Society, ou_1753326              

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Free keywords: synthesis inhibitory factor virus protein bcrf1 endoplasmic-reticulum transgenic plants crystal-structure monoclonal-antibody mass-spectrometry human neutrophils interferon-gamma golgi-apparatus
 Abstract: Background: Interleukin-10 (IL-10) is a potent anti-inflammatory cytokine, with therapeutic applications in several autoimmune and inflammatory diseases. Oral administration of this cytokine alone, or in combination with disease-associated autoantigens could confer protection form the onset of a specific autoimmune disease through the induction of oral tolerance. Transgenic plants are attractive systems for production of therapeutic proteins because of the ability to do large scale-up at low cost, and the low maintenance requirements. They are highly amenable to oral administration and could become effective delivery systems without extensive protein purification. We investigated the ability of tobacco plants to produce high levels of biologically-active viral and murine IL-10. Results: Three different subcellular targeting strategies were assessed in transient expression experiments, and stable transgenic tobacco plants were generated with the constructs that yielded the highest accumulation levels by targeting the recombinant proteins to the endoplasmic reticulum. The best yields using this strategy in T-1 plants were 10.8 and 37.0 mu g/g fresh leaf weight for viral and murine IL-10, respectively. The recombinant proteins were purified from transgenic leaf material and characterized in terms of their N-glycan composition, dimerization and biological activity in in vitro assays. Both molecules formed stable dimers, were able to activate the IL-10 signaling pathway and to induce specific anti-inflammatory responses in mouse J774 macrophage cells. Conclusion: Tobacco plants are able to correctly process viral and murine IL-10 into biologically active dimers, therefore representing a suitable platform for the production for these cytokines. The accumulation levels obtained are high enough to allow delivery of an immunologically relevant dose of IL-10 in a reasonable amount of leaf material, without extensive purification. This study paves the way to performing feeding studies in mouse models of autoimmune diseases, that will allow the evaluation the immunomodulatory properties and effectiveness of the viral IL-10 in inducing oral tolerance compared to the murine protein.

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Language(s): eng - English
 Dates: 2009-03-192009
 Publication Status: Published in print
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 Table of Contents: -
 Rev. Type: -
 Identifiers: ISI: ISI:000265608100001
DOI: 10.1186/1472-6750-9-22
ISSN: 1472-6750 (Electronic) 1472-6750 (Linking)
URI: ://000265608100001 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2667500/pdf/1472-6750-9-22.pdf?tool=pmcentrez
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Title: BMC Biotechnology
Source Genre: Journal
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Pages: - Volume / Issue: 9 Sequence Number: - Start / End Page: 22 Identifier: -