Past Issue

Volume 16, Number 3, Autumn 2014, Serial Number: 63, Pages: 255-262

Lentiviral Mediating Genetic Engineered Mesenchymal Stem Cells for Releasing IL-27 as a Gene Therapy Approach for Autoimmune Diseases


Shohreh Hajizadeh-Sikaroodi, Ph.D, 1, Ahmad Hosseini, Ph.D, 2, 3, 4, *, Ali Fallah, M.Sc, 5, Hajar Estiri, M.Sc, 6, Zahra Noormohammadi, Ph.D, 7, Mohammad Salehi, Ph.D, 2, 3, Sayyed Mohammad Hossein Ghaderian, Ph.D, 8, Haleh Akhavan Niaki, Ph.D, 9, Masoud Soleimani, Ph.D, 10, Bahram Kazemi, Ph.D, 2, 11,
Science and Research Branch, Islamic Azad University, Tehran, Iran
Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Mehr Infertility Research Center, Rasht, Iran
Department of Cell Biology and Anatomical Science, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Systems and Synthetic Biology Group, Mede Bioeconomy Company, Tehran, Iran
Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, Tehran, Iran
Department of Biology, Faculty of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran
Department of Medical Genetics, Faculty of Medicine, Shahid Beheshti University of Medical Sciences and Health Services, Tehran, Iran
Cellular and Molecular Biology Research Center, Babol University of Medical Sciences, Babol, Iran
Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Department of Biotechnology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
*Corresponding Address: P.O.Box: 193954719 Cellular and Molecular Biology Research Center Shahid Beheshti University of Medical Sciences Tehran Iran Email:Prof_hosseini@yahoo.com

Abstract

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-IRES- EBI3-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.