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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 the p...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Royan Institute
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4204184/ https://www.ncbi.nlm.nih.gov/pubmed/24611150 |
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author | Hajizadeh-Sikaroodi, Shohreh Hosseini, Ahmad Fallah, Ali Estiri, Hajar Noormohammadi, Zahra Salehi, Mohammad Ghaderian, Sayyed Mohammad Hossein Akhavan Niaki, Haleh Soleimani, Masoud Kazemi, Bahram |
author_facet | Hajizadeh-Sikaroodi, Shohreh Hosseini, Ahmad Fallah, Ali Estiri, Hajar Noormohammadi, Zahra Salehi, Mohammad Ghaderian, Sayyed Mohammad Hossein Akhavan Niaki, Haleh Soleimani, Masoud Kazemi, Bahram |
author_sort | Hajizadeh-Sikaroodi, Shohreh |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4204184 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Royan Institute |
record_format | MEDLINE/PubMed |
spelling | pubmed-42041842014-11-07 Lentiviral Mediating Genetic Engineered Mesenchymal Stem Cells for Releasing IL-27 as a Gene Therapy Approach for Autoimmune Diseases Hajizadeh-Sikaroodi, Shohreh Hosseini, Ahmad Fallah, Ali Estiri, Hajar Noormohammadi, Zahra Salehi, Mohammad Ghaderian, Sayyed Mohammad Hossein Akhavan Niaki, Haleh Soleimani, Masoud Kazemi, Bahram Cell J Original Article 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. Royan Institute 2014 2014-10-04 /pmc/articles/PMC4204184/ /pubmed/24611150 Text en Any use, distribution, reproduction or abstract of this publication in any medium, with the exception of commercial purposes, is permitted provided the original work is properly cited http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Article Hajizadeh-Sikaroodi, Shohreh Hosseini, Ahmad Fallah, Ali Estiri, Hajar Noormohammadi, Zahra Salehi, Mohammad Ghaderian, Sayyed Mohammad Hossein Akhavan Niaki, Haleh Soleimani, Masoud Kazemi, Bahram Lentiviral Mediating Genetic Engineered Mesenchymal Stem Cells for Releasing IL-27 as a Gene Therapy Approach for Autoimmune Diseases |
title | Lentiviral Mediating Genetic Engineered Mesenchymal
Stem Cells for Releasing IL-27 as a Gene Therapy
Approach for Autoimmune Diseases |
title_full | Lentiviral Mediating Genetic Engineered Mesenchymal
Stem Cells for Releasing IL-27 as a Gene Therapy
Approach for Autoimmune Diseases |
title_fullStr | Lentiviral Mediating Genetic Engineered Mesenchymal
Stem Cells for Releasing IL-27 as a Gene Therapy
Approach for Autoimmune Diseases |
title_full_unstemmed | Lentiviral Mediating Genetic Engineered Mesenchymal
Stem Cells for Releasing IL-27 as a Gene Therapy
Approach for Autoimmune Diseases |
title_short | Lentiviral Mediating Genetic Engineered Mesenchymal
Stem Cells for Releasing IL-27 as a Gene Therapy
Approach for Autoimmune Diseases |
title_sort | lentiviral mediating genetic engineered mesenchymal
stem cells for releasing il-27 as a gene therapy
approach for autoimmune diseases |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4204184/ https://www.ncbi.nlm.nih.gov/pubmed/24611150 |
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