highly efficient stable expression of indoleamine 2,3 dioxygenase gene in primary fibroblasts高效稳定表达的吲哚胺2,3加双氧酶基因主要的成纤维细胞.pdfVIP

Biol Proced Online (2010) 12:107– 112 DOI 10.1007/s12575-010-9028-6 Highly Efficient Stable Expression of Indoleamine 2,3 Dioxygenase Gene in Primary Fibroblasts Alireza Moeen Rezakhanlou Darya Habibi Amy Lai Reza B. Jalili Christopher J. Ong Aziz Ghahary Received: 3 September 2009 /Accepted: 20 February 2010 /Published online: 27 March 2010 # The Author(s) 2010. This article is published with open access at S Abstract Indoleamine 2,3 dioxygenase (IDO) is a potent fibroblasts and showed a significant increase (about tenfold) immunomodulatory enzyme that has recently attracted in the rate of gene transfection. In addition, by the use of significant attention for its potential application as an fluorescence-activated cell sorting, a 95% pure population inducer of immunotolerance in transplantation. We have of IDO-expressing fibroblasts was successfully obtained. previously demonstrated that a collagen matrix populated The efficiency of the IDO expression and the activity of with IDO-expressing fibroblasts can be applied successfully the enzyme have been confirmed by Western blotting, in suppressing islet allogeneic immune response. Meanwhile, fluorescence-activated cell sorting analysis, and Kynurenine a critical aspect of such immunological intervention relies assay, respectively. The findings of this study revealed largely on effective long-term expression of the IDO gene. simple and effective strategies through which an efficient Moreover, gene manipulation of primary cells is known to be and stable expression of IDO can be achieved for primary challenging due to unsatisfactory expression of the exogenous cells which, in turn, significantly improves its potential as gene. In this