Highly Efficient Stable Expression of Indoleamine 2,3 Dioxygenase Gene in Primary Fibroblasts
<p>Abstract</p> <p>Indoleamine 2,3 dioxygenase (IDO) is a potent immunomodulatory enzyme that has recently attracted significant attention for its potential application as an inducer of immunotolerance in transplantation. We have previously demonstrated that a collagen matrix popul...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2010-01-01
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| Series: | Biological Procedures Online |
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| Online Access: | http://www.biologicalproceduresonline.com/content/12/1/107 |
| _version_ | 1811331216817782784 |
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| author | Ong Christopher Rezakhanlou Alireza Habibi Darya Lai Amy Jalili Reza Ghahary Aziz |
| author_facet | Ong Christopher Rezakhanlou Alireza Habibi Darya Lai Amy Jalili Reza Ghahary Aziz |
| author_sort | Ong Christopher |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Indoleamine 2,3 dioxygenase (IDO) is a potent immunomodulatory enzyme that has recently attracted significant attention for its potential application as an inducer of immunotolerance in transplantation. We have previously demonstrated that a collagen matrix populated with IDO-expressing fibroblasts can be applied successfully in suppressing islet allogeneic immune response. Meanwhile, a critical aspect of such immunological intervention relies largely on effective long-term expression of the IDO gene. Moreover, gene manipulation of primary cells is known to be challenging due to unsatisfactory expression of the exogenous gene. In this study, a lentiviral gene delivery system has been employed to transduce primary fibroblasts. We used polybrene to efficiently deliver the IDO gene into primary fibroblasts and showed a significant increase (about tenfold) in the rate of gene transfection. In addition, by the use of fluorescence-activated cell sorting, a 95% pure population of IDO-expressing fibroblasts was successfully obtained. The efficiency of the IDO expression and the activity of the enzyme have been confirmed by Western blotting, fluorescence-activated cell sorting analysis, and Kynurenine assay, respectively. The findings of this study revealed simple and effective strategies through which an efficient and stable expression of IDO can be achieved for primary cells which, in turn, significantly improves its potential as a tool for achieving immunotolerance in different types of transplantation.</p> |
| first_indexed | 2024-04-13T16:15:41Z |
| format | Article |
| id | doaj.art-5ac60780482b4ce1ab260d6bb8584ddb |
| institution | Directory Open Access Journal |
| issn | 1480-9222 |
| language | English |
| last_indexed | 2024-04-13T16:15:41Z |
| publishDate | 2010-01-01 |
| publisher | BMC |
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| spelling | doaj.art-5ac60780482b4ce1ab260d6bb8584ddb2022-12-22T02:40:03ZengBMCBiological Procedures Online1480-92222010-01-01121107112Highly Efficient Stable Expression of Indoleamine 2,3 Dioxygenase Gene in Primary FibroblastsOng ChristopherRezakhanlou AlirezaHabibi DaryaLai AmyJalili RezaGhahary Aziz<p>Abstract</p> <p>Indoleamine 2,3 dioxygenase (IDO) is a potent immunomodulatory enzyme that has recently attracted significant attention for its potential application as an inducer of immunotolerance in transplantation. We have previously demonstrated that a collagen matrix populated with IDO-expressing fibroblasts can be applied successfully in suppressing islet allogeneic immune response. Meanwhile, a critical aspect of such immunological intervention relies largely on effective long-term expression of the IDO gene. Moreover, gene manipulation of primary cells is known to be challenging due to unsatisfactory expression of the exogenous gene. In this study, a lentiviral gene delivery system has been employed to transduce primary fibroblasts. We used polybrene to efficiently deliver the IDO gene into primary fibroblasts and showed a significant increase (about tenfold) in the rate of gene transfection. In addition, by the use of fluorescence-activated cell sorting, a 95% pure population of IDO-expressing fibroblasts was successfully obtained. The efficiency of the IDO expression and the activity of the enzyme have been confirmed by Western blotting, fluorescence-activated cell sorting analysis, and Kynurenine assay, respectively. The findings of this study revealed simple and effective strategies through which an efficient and stable expression of IDO can be achieved for primary cells which, in turn, significantly improves its potential as a tool for achieving immunotolerance in different types of transplantation.</p>http://www.biologicalproceduresonline.com/content/12/1/107Lentiviral vectorIndoleamine 23 dioxygenasePrimary fibroblastTransplantationImmunogenicity |
| spellingShingle | Ong Christopher Rezakhanlou Alireza Habibi Darya Lai Amy Jalili Reza Ghahary Aziz Highly Efficient Stable Expression of Indoleamine 2,3 Dioxygenase Gene in Primary Fibroblasts Biological Procedures Online Lentiviral vector Indoleamine 2 3 dioxygenase Primary fibroblast Transplantation Immunogenicity |
| title | Highly Efficient Stable Expression of Indoleamine 2,3 Dioxygenase Gene in Primary Fibroblasts |
| title_full | Highly Efficient Stable Expression of Indoleamine 2,3 Dioxygenase Gene in Primary Fibroblasts |
| title_fullStr | Highly Efficient Stable Expression of Indoleamine 2,3 Dioxygenase Gene in Primary Fibroblasts |
| title_full_unstemmed | Highly Efficient Stable Expression of Indoleamine 2,3 Dioxygenase Gene in Primary Fibroblasts |
| title_short | Highly Efficient Stable Expression of Indoleamine 2,3 Dioxygenase Gene in Primary Fibroblasts |
| title_sort | highly efficient stable expression of indoleamine 2 3 dioxygenase gene in primary fibroblasts |
| topic | Lentiviral vector Indoleamine 2 3 dioxygenase Primary fibroblast Transplantation Immunogenicity |
| url | http://www.biologicalproceduresonline.com/content/12/1/107 |
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