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Adenoviral transduction of mesenchymal stem cells: in vitro responses and in vivo immune responses after cell transplantation.

Treacy O, Ryan AE, Heinzl T, O'Flynn L, Cregg M, Wilk M, Odoardi F, Lohan P, O'Brien T, Nosov M, Ritter T - PLoS ONE (2012)

Bottom Line: We found that Ad-transduction of MSCs does not lead to up-regulation of major histocompatibility complex class I and II and co-stimulatory molecules CD80 and CD86.In vivo injection of Ad-transduced MSCs did not change the frequency of various immune cell populations (antigen presenting cells, T helper and cytotoxic T cells, natural killer and natural killer T cells) neither in the blood nor in tissues.Our results indicate that Ad-modification has no major influence on the immunological properties of MSCs and therefore can be considered as a suitable gene vector for therapeutic applications of MSCs.

View Article: PubMed Central - PubMed

Affiliation: College of Medicine, Nursing and Health Sciences, School of Medicine, Regenerative Medicine Institute, National University of Ireland, Galway, Ireland.

ABSTRACT
Adult mesenchymal stem cells (MSCs) are non-hematopoietic cells with multi-lineage potential which makes them attractive targets for regenerative medicine applications. However, to date, therapeutic success of MSC-therapy is limited and the genetic modification of MSCs using viral vectors is one option to improve their therapeutic potential. Ex-vivo genetic modification of MSCs using recombinant adenovirus (Ad) could be promising to reduce undesired immune responses as Ad will be removed before cell/tissue transplantation. In this regard, we investigated whether Ad-modification of MSCs alters their immunological properties in vitro and in vivo. We found that Ad-transduction of MSCs does not lead to up-regulation of major histocompatibility complex class I and II and co-stimulatory molecules CD80 and CD86. Moreover, Ad-transduction caused no significant changes in terms of pro-inflammatory cytokine expression, chemokine and chemokine receptor and Toll-like receptor expression. In addition, Ad-modification of MSCs had no affect on their ability to suppress T cell proliferation in vitro. In vivo injection of Ad-transduced MSCs did not change the frequency of various immune cell populations (antigen presenting cells, T helper and cytotoxic T cells, natural killer and natural killer T cells) neither in the blood nor in tissues. Our results indicate that Ad-modification has no major influence on the immunological properties of MSCs and therefore can be considered as a suitable gene vector for therapeutic applications of MSCs.

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Related in: MedlinePlus

Analysis of immune cell populations after injection of untransduced or Ad-transduced MSCs.The percentages of (A) CD4+CD25+, (B) CD11b/c+MHCII+, (C) CD3+CD8+CD161+, (D) CD3+CD8+CD161++, (E) CD3+CD8+CD161− and (F) CD3−CD8+CD161++ cells in the PBMC population at days 3, 7 and 14, and also in the lung and spleen at day 14 post-injection of either 2×106 Ad.GFP transduced MSCs (n = 4) or 2×106 untransduced MSCs (n = 3) as measured by flow cytometry. (G) The percentage of HIS36+ cells in the lung and spleen at day 14 post-injection as measured by flow cytometry. Statistical analysis was performed using the non-parametric Mann-Whitney test. For all tested parameters, p values were >0.05.
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pone-0042662-g006: Analysis of immune cell populations after injection of untransduced or Ad-transduced MSCs.The percentages of (A) CD4+CD25+, (B) CD11b/c+MHCII+, (C) CD3+CD8+CD161+, (D) CD3+CD8+CD161++, (E) CD3+CD8+CD161− and (F) CD3−CD8+CD161++ cells in the PBMC population at days 3, 7 and 14, and also in the lung and spleen at day 14 post-injection of either 2×106 Ad.GFP transduced MSCs (n = 4) or 2×106 untransduced MSCs (n = 3) as measured by flow cytometry. (G) The percentage of HIS36+ cells in the lung and spleen at day 14 post-injection as measured by flow cytometry. Statistical analysis was performed using the non-parametric Mann-Whitney test. For all tested parameters, p values were >0.05.

Mentions: In order to understand if genetic modification of MSCs using Ad-vectors may induce an inflammatory response in vivo, MSCs were transduced ex-vivo with Ad.GFP (MOI 100) and injected intravenously (i.v.) into syngeneic CD rats (2×106 cells/animal (n = 4). Untransduced MSCs (2×106 cells/animal) were injected as control (n = 3). Blood samples of treated animals were taken on days 3, 7 and on the day of sacrifice (day 14) and the frequency of several blood cell populations (antigen presenting cells, CD11b/c+MHCII+; activated T cells and cytotoxic T cells, CD4+CD25+ and CD3+CD8+CD161−; activated natural killer cells, CD3−CD8+CD161++ and natural killer T cells CD3+CD8+CD161+, CD3+CD8+CD161++) were analysed by flow cytometry. Moreover, spleens and lungs from animals treated with either untransduced or Ad.GFP-transduced MSCs were collected and analysed for the presence of the same immune cell populations as for blood. Additionally, we stained for HIS36 expression as evidence of tissue macrophage presence in lungs and spleens of treated animals on the day of sacrifice. No significant differences in the percentage and in the activation status of the different blood cell populations analysed was observed on days 3, 7 and 14 (Figure 6A–F and figure S3A for gating strategy) after injection of either untransduced or Ad.GFP-transduced MSCs. Moreover, similar results were observed when analysing the immune cell populations isolated from the lungs and spleens of both treatment groups (Figure 6A–G and figure S3B and C).


Adenoviral transduction of mesenchymal stem cells: in vitro responses and in vivo immune responses after cell transplantation.

Treacy O, Ryan AE, Heinzl T, O'Flynn L, Cregg M, Wilk M, Odoardi F, Lohan P, O'Brien T, Nosov M, Ritter T - PLoS ONE (2012)

Analysis of immune cell populations after injection of untransduced or Ad-transduced MSCs.The percentages of (A) CD4+CD25+, (B) CD11b/c+MHCII+, (C) CD3+CD8+CD161+, (D) CD3+CD8+CD161++, (E) CD3+CD8+CD161− and (F) CD3−CD8+CD161++ cells in the PBMC population at days 3, 7 and 14, and also in the lung and spleen at day 14 post-injection of either 2×106 Ad.GFP transduced MSCs (n = 4) or 2×106 untransduced MSCs (n = 3) as measured by flow cytometry. (G) The percentage of HIS36+ cells in the lung and spleen at day 14 post-injection as measured by flow cytometry. Statistical analysis was performed using the non-parametric Mann-Whitney test. For all tested parameters, p values were >0.05.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3412834&req=5

pone-0042662-g006: Analysis of immune cell populations after injection of untransduced or Ad-transduced MSCs.The percentages of (A) CD4+CD25+, (B) CD11b/c+MHCII+, (C) CD3+CD8+CD161+, (D) CD3+CD8+CD161++, (E) CD3+CD8+CD161− and (F) CD3−CD8+CD161++ cells in the PBMC population at days 3, 7 and 14, and also in the lung and spleen at day 14 post-injection of either 2×106 Ad.GFP transduced MSCs (n = 4) or 2×106 untransduced MSCs (n = 3) as measured by flow cytometry. (G) The percentage of HIS36+ cells in the lung and spleen at day 14 post-injection as measured by flow cytometry. Statistical analysis was performed using the non-parametric Mann-Whitney test. For all tested parameters, p values were >0.05.
Mentions: In order to understand if genetic modification of MSCs using Ad-vectors may induce an inflammatory response in vivo, MSCs were transduced ex-vivo with Ad.GFP (MOI 100) and injected intravenously (i.v.) into syngeneic CD rats (2×106 cells/animal (n = 4). Untransduced MSCs (2×106 cells/animal) were injected as control (n = 3). Blood samples of treated animals were taken on days 3, 7 and on the day of sacrifice (day 14) and the frequency of several blood cell populations (antigen presenting cells, CD11b/c+MHCII+; activated T cells and cytotoxic T cells, CD4+CD25+ and CD3+CD8+CD161−; activated natural killer cells, CD3−CD8+CD161++ and natural killer T cells CD3+CD8+CD161+, CD3+CD8+CD161++) were analysed by flow cytometry. Moreover, spleens and lungs from animals treated with either untransduced or Ad.GFP-transduced MSCs were collected and analysed for the presence of the same immune cell populations as for blood. Additionally, we stained for HIS36 expression as evidence of tissue macrophage presence in lungs and spleens of treated animals on the day of sacrifice. No significant differences in the percentage and in the activation status of the different blood cell populations analysed was observed on days 3, 7 and 14 (Figure 6A–F and figure S3A for gating strategy) after injection of either untransduced or Ad.GFP-transduced MSCs. Moreover, similar results were observed when analysing the immune cell populations isolated from the lungs and spleens of both treatment groups (Figure 6A–G and figure S3B and C).

Bottom Line: We found that Ad-transduction of MSCs does not lead to up-regulation of major histocompatibility complex class I and II and co-stimulatory molecules CD80 and CD86.In vivo injection of Ad-transduced MSCs did not change the frequency of various immune cell populations (antigen presenting cells, T helper and cytotoxic T cells, natural killer and natural killer T cells) neither in the blood nor in tissues.Our results indicate that Ad-modification has no major influence on the immunological properties of MSCs and therefore can be considered as a suitable gene vector for therapeutic applications of MSCs.

View Article: PubMed Central - PubMed

Affiliation: College of Medicine, Nursing and Health Sciences, School of Medicine, Regenerative Medicine Institute, National University of Ireland, Galway, Ireland.

ABSTRACT
Adult mesenchymal stem cells (MSCs) are non-hematopoietic cells with multi-lineage potential which makes them attractive targets for regenerative medicine applications. However, to date, therapeutic success of MSC-therapy is limited and the genetic modification of MSCs using viral vectors is one option to improve their therapeutic potential. Ex-vivo genetic modification of MSCs using recombinant adenovirus (Ad) could be promising to reduce undesired immune responses as Ad will be removed before cell/tissue transplantation. In this regard, we investigated whether Ad-modification of MSCs alters their immunological properties in vitro and in vivo. We found that Ad-transduction of MSCs does not lead to up-regulation of major histocompatibility complex class I and II and co-stimulatory molecules CD80 and CD86. Moreover, Ad-transduction caused no significant changes in terms of pro-inflammatory cytokine expression, chemokine and chemokine receptor and Toll-like receptor expression. In addition, Ad-modification of MSCs had no affect on their ability to suppress T cell proliferation in vitro. In vivo injection of Ad-transduced MSCs did not change the frequency of various immune cell populations (antigen presenting cells, T helper and cytotoxic T cells, natural killer and natural killer T cells) neither in the blood nor in tissues. Our results indicate that Ad-modification has no major influence on the immunological properties of MSCs and therefore can be considered as a suitable gene vector for therapeutic applications of MSCs.

Show MeSH
Related in: MedlinePlus