Lentiviral Mediating Genetic Engineered Mesenchymal Stem Cells for Releasing IL-27 as a Gene Therapy Approach for Autoimmune Diseases
Objective: Autoimmune diseases precede a complex dysregulation of the immune system. T helper17 (Th17) and interleukin (IL)-17 have central roles in initiation of inflammation and subsequent autoimmune diseases. IL-27 significantly controls autoimmune diseases by Th17 and IL-17 suppression. In th...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Royan Institute (ACECR), Tehran
2014-04-01
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| Series: | Cell Journal |
| Subjects: | |
| Online Access: | http://celljournal.org/library/upload/article/af_25234445425447253433253326544324527635323-Shohreh%20Hajizadeh-1-1-phone.pdf |
| Summary: | Objective: Autoimmune diseases precede a complex dysregulation of the immune system.
T helper17 (Th17) and interleukin (IL)-17 have central roles in initiation of inflammation
and subsequent autoimmune diseases. IL-27 significantly controls autoimmune
diseases by Th17 and IL-17 suppression. In the present study we have created genetic
engineered mesenchymal stem cells (MSCs) that mediate with lentiviral vectors to release
IL-27 as an adequate vehicle for ex vivo gene therapy in the reduction of inflammation and
autoimmune diseases.
Materials and Methods: In this experimental study, we isolated adipose-derived MSCs
(AD-MSCs) from lipoaspirate and subsequently characterized them by differentiation. Two
subunits of IL-27 (p28 and EBI3) were cloned in a pCDH-513B-1 lentiviral vector. Expressions
of p28 and EBI3 (Epstein-Barr virus induced gene 3) were determined by real time
polymerase chain reaction (PCR). MSCs were transduced by a pCDH-CMV-p28-IRESEBI3-
EF-copGFP-Pur lentiviral vector and the bioassay of IL-27 was evaluated by IL-10
expression.
Results: Cell differentiation confirmed true isolation of MSCs from lipoaspirate. Restriction
enzyme digestion and sequencing verified successful cloning of both p28
and EBI3 in the pCDH-513B-1 lentiviral vector. Real time PCR showed high expressions
level of IL-27 and IL-10 as well as accurate activity of IL-27.
Conclusion: The results showed transduction of functional IL-27 to AD-MSCs by means of a
lentiviral vector. The lentiviral vector did not impact MSC characteristics. |
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| ISSN: | 2228-5806 2228-5814 |