Evaluation of gene delivery strategies to efficiently overexpress functional HLA-G on human bone marrow stromal cells
Mesenchymal stromal cells (MSC) constitutively express low levels of human leukocyte antigen-G (HLA-G), which has been shown to contribute to their immunomodulatory and anti-inflammatory properties. Here, we hypothesized that overexpression of HLA-G on bone marrow-derived MSC would improve their imm...
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doaj-1a4674f592854288b2ca80ff049638c12020-11-25T00:21:40ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012014-01-011C10.1038/mtm.2014.41Evaluation of gene delivery strategies to efficiently overexpress functional HLA-G on human bone marrow stromal cellsJoana S Boura0Melisa Vance1Weihong Yin2Catarina Madeira3Cláudia Lobato da Silva4Christopher D Porada5Graça Almeida-Porada6Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, North Carolina, USAWake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, North Carolina, USAWake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, North Carolina, USAInstitute for Biotechnology and Bioengineering and Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, PortugalInstitute for Biotechnology and Bioengineering and Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, PortugalWake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, North Carolina, USAWake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, North Carolina, USAMesenchymal stromal cells (MSC) constitutively express low levels of human leukocyte antigen-G (HLA-G), which has been shown to contribute to their immunomodulatory and anti-inflammatory properties. Here, we hypothesized that overexpression of HLA-G on bone marrow-derived MSC would improve their immunomodulatory function, thus increasing their therapeutic potential. Therefore, we investigated which gene transfer system is best suited for delivering this molecule while maintaining its immunomodulatory effects. We performed a side-by-side comparison between three nonviral plasmid-based platforms (pmax-HLA-G1; MC-HLA-G1; pEP-HLA-G1) and a viral system (Lv-HLA-G1) using gene transfer parameters that yielded similar levels of HLA-G1-expressing MSC. Natural killer (NK) cell–mediated lysis assays and T cell proliferation assays showed that MSC modified with the HLA-G1 expressing viral vector had significantly lower susceptibility to NK-lysis and significantly reduced T cell proliferation when compared to nonmodified cells or MSC modified with plasmid. We also show that, in plasmid-modified MSC, an increase in Toll-like receptor (TLR)9 expression is the mechanism responsible for the abrogation of HLA-G1's immunomodulatory effect. Although MSC can be efficiently modified to overexpress HLA-G1 using viral and nonviral strategies, only viral-based delivery of HLA-G1 is suitable for improvement of MSC's immunomodulatory properties.http://www.sciencedirect.com/science/article/pii/S2329050116301097 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Joana S Boura Melisa Vance Weihong Yin Catarina Madeira Cláudia Lobato da Silva Christopher D Porada Graça Almeida-Porada |
spellingShingle |
Joana S Boura Melisa Vance Weihong Yin Catarina Madeira Cláudia Lobato da Silva Christopher D Porada Graça Almeida-Porada Evaluation of gene delivery strategies to efficiently overexpress functional HLA-G on human bone marrow stromal cells Molecular Therapy: Methods & Clinical Development |
author_facet |
Joana S Boura Melisa Vance Weihong Yin Catarina Madeira Cláudia Lobato da Silva Christopher D Porada Graça Almeida-Porada |
author_sort |
Joana S Boura |
title |
Evaluation of gene delivery strategies to efficiently overexpress functional HLA-G on human bone marrow stromal cells |
title_short |
Evaluation of gene delivery strategies to efficiently overexpress functional HLA-G on human bone marrow stromal cells |
title_full |
Evaluation of gene delivery strategies to efficiently overexpress functional HLA-G on human bone marrow stromal cells |
title_fullStr |
Evaluation of gene delivery strategies to efficiently overexpress functional HLA-G on human bone marrow stromal cells |
title_full_unstemmed |
Evaluation of gene delivery strategies to efficiently overexpress functional HLA-G on human bone marrow stromal cells |
title_sort |
evaluation of gene delivery strategies to efficiently overexpress functional hla-g on human bone marrow stromal cells |
publisher |
Elsevier |
series |
Molecular Therapy: Methods & Clinical Development |
issn |
2329-0501 |
publishDate |
2014-01-01 |
description |
Mesenchymal stromal cells (MSC) constitutively express low levels of human leukocyte antigen-G (HLA-G), which has been shown to contribute to their immunomodulatory and anti-inflammatory properties. Here, we hypothesized that overexpression of HLA-G on bone marrow-derived MSC would improve their immunomodulatory function, thus increasing their therapeutic potential. Therefore, we investigated which gene transfer system is best suited for delivering this molecule while maintaining its immunomodulatory effects. We performed a side-by-side comparison between three nonviral plasmid-based platforms (pmax-HLA-G1; MC-HLA-G1; pEP-HLA-G1) and a viral system (Lv-HLA-G1) using gene transfer parameters that yielded similar levels of HLA-G1-expressing MSC. Natural killer (NK) cell–mediated lysis assays and T cell proliferation assays showed that MSC modified with the HLA-G1 expressing viral vector had significantly lower susceptibility to NK-lysis and significantly reduced T cell proliferation when compared to nonmodified cells or MSC modified with plasmid. We also show that, in plasmid-modified MSC, an increase in Toll-like receptor (TLR)9 expression is the mechanism responsible for the abrogation of HLA-G1's immunomodulatory effect. Although MSC can be efficiently modified to overexpress HLA-G1 using viral and nonviral strategies, only viral-based delivery of HLA-G1 is suitable for improvement of MSC's immunomodulatory properties. |
url |
http://www.sciencedirect.com/science/article/pii/S2329050116301097 |
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