Lentiviral Mediating Genetic Engineered Mesenchymal Stem Cells for Releasing IL-27 as a Gene Therapy Approach for Autoimmune Diseases
Shohreh Hajizadeh-Sikaroodi,
Ahmad Hosseini,
Ali Falla,
Hajar Estiri,
Zahra Noormohammadi,
Mohammad Salehi,
Sayyed Mohammad Hossein Ghaderian,
Haleh Akhavan Niaki,
Masoud Soleimani,
Bahram Kazemi
Affiliations
Shohreh Hajizadeh-Sikaroodi
Science and Research Branch, Islamic Azad University, Tehran, Iran
Ahmad Hosseini
Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran and Mehr Infertility Research Center, Rasht, Iran and Department of Cell Biology and Anatomical Science, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Ali Falla
Systems and Synthetic Biology Group, Mede Bioeconomy Company, Tehran, Iran
Hajar Estiri
Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, Tehran, Iran
Zahra Noormohammadi
Department of Biology, Faculty of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran
Mohammad Salehi
Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran and Mehr Infertility Research Center, Rasht, Iran
Sayyed Mohammad Hossein Ghaderian
Department of Medical Genetics, Faculty of Medicine, Shahid Beheshti University of Medical Sciences and Health Services, Tehran, Iran
Haleh Akhavan Niaki
Cellular and Molecular Biology Research Center, Babol University of Medical Sciences, Babol, Iran
Masoud Soleimani
Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Bahram Kazemi
Department of Biotechnology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran and Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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.
