title: Lentiviral Mediating Genetic Engineered Mesenchymal Stem Cells for Releasing IL-27 as a Gene Therapy Approach for Autoimmune Diseases.
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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Shohreh
authors.initials: S
authors.lastname: Hajizadeh-Sikaroodi
authors.affiliation: 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.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Hosseini
authors.affiliation: Systems and Synthetic Biology Group, Mede Bioeconomy Company, Tehran, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Fallah
authors.affiliation: Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, Tehran, Iran.
authors.forename: Hajar
authors.initials: H
authors.lastname: Estiri
authors.affiliation: Department of Biology, Faculty of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Noormohammadi
authors.affiliation: Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; Mehr Infertility Research Center, Rasht, Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Salehi
authors.affiliation: Department of Medical Genetics, Faculty of Medicine, Shahid Beheshti University of Medical Sciences and Health Services, Tehran, Iran.
authors.forename: Sayyed Mohammad Hossein
authors.initials: SM
authors.lastname: Ghaderian
authors.affiliation: Cellular and Molecular Biology Research Center, Babol University of Medical Sciences, Babol, Iran.
authors.forename: Haleh
authors.initials: H
authors.lastname: Akhavan Niaki
authors.affiliation: Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.affiliation: Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; Department of Biotechnology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Bahram
authors.initials: B
authors.lastname: Kazemi
keywords: Autoimmune Disease; Gene Therapy; IL-27; Mesenchymal Stem Cells