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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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T-cell proliferation in the presence or absence of untransduced and Ad.GFP transduced MSCs.(A) Histograms showing percent proliferation of CFSE-labelled T cells that were polyclonally stimulated with anti-CD3/anti-CD28 beads in the absence or presence of either untransduced or Ad.GFP transduced MSCs (ratio of T cells to MSCs; 1∶250). (B) Bar chart showing the percentage of cells divided greater than three generations following polyclonal stimulation in the absence or presence of either untransduced or Ad.GFP transduced MSCs. CFSE fluorescence was analysed on day 4 after co-culture. Shown is a representative experiment with means of 4 replicates ± SD (**: p<0.01; student's T test).
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pone-0042662-g005: T-cell proliferation in the presence or absence of untransduced and Ad.GFP transduced MSCs.(A) Histograms showing percent proliferation of CFSE-labelled T cells that were polyclonally stimulated with anti-CD3/anti-CD28 beads in the absence or presence of either untransduced or Ad.GFP transduced MSCs (ratio of T cells to MSCs; 1∶250). (B) Bar chart showing the percentage of cells divided greater than three generations following polyclonal stimulation in the absence or presence of either untransduced or Ad.GFP transduced MSCs. CFSE fluorescence was analysed on day 4 after co-culture. Shown is a representative experiment with means of 4 replicates ± SD (**: p<0.01; student's T test).

Mentions: Finally we investigated whether Ad-transduction of MSCs alters the capacity of MSCs to modulate T cell proliferation. For this, polyclonally activated T cells (stimulated with anti-CD3/anti-CD28 coated beads) were co-cultured with either untransduced or Ad.GFP-transduced MSCs. T cells were labelled with CFSE and after a 4-day co-culture with MSCs, collected and subjected to flow cytometric analysis. We found that co-culture of MSCs with polyclonally activated T cells significantly inhibited the proliferation of CFSE-labelled T cells and this was not altered upon genetic modification of MSCs using Ad-vectors (p<0.01, Figure 5A and B).


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)

T-cell proliferation in the presence or absence of untransduced and Ad.GFP transduced MSCs.(A) Histograms showing percent proliferation of CFSE-labelled T cells that were polyclonally stimulated with anti-CD3/anti-CD28 beads in the absence or presence of either untransduced or Ad.GFP transduced MSCs (ratio of T cells to MSCs; 1∶250). (B) Bar chart showing the percentage of cells divided greater than three generations following polyclonal stimulation in the absence or presence of either untransduced or Ad.GFP transduced MSCs. CFSE fluorescence was analysed on day 4 after co-culture. Shown is a representative experiment with means of 4 replicates ± SD (**: p<0.01; student's T test).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0042662-g005: T-cell proliferation in the presence or absence of untransduced and Ad.GFP transduced MSCs.(A) Histograms showing percent proliferation of CFSE-labelled T cells that were polyclonally stimulated with anti-CD3/anti-CD28 beads in the absence or presence of either untransduced or Ad.GFP transduced MSCs (ratio of T cells to MSCs; 1∶250). (B) Bar chart showing the percentage of cells divided greater than three generations following polyclonal stimulation in the absence or presence of either untransduced or Ad.GFP transduced MSCs. CFSE fluorescence was analysed on day 4 after co-culture. Shown is a representative experiment with means of 4 replicates ± SD (**: p<0.01; student's T test).
Mentions: Finally we investigated whether Ad-transduction of MSCs alters the capacity of MSCs to modulate T cell proliferation. For this, polyclonally activated T cells (stimulated with anti-CD3/anti-CD28 coated beads) were co-cultured with either untransduced or Ad.GFP-transduced MSCs. T cells were labelled with CFSE and after a 4-day co-culture with MSCs, collected and subjected to flow cytometric analysis. We found that co-culture of MSCs with polyclonally activated T cells significantly inhibited the proliferation of CFSE-labelled T cells and this was not altered upon genetic modification of MSCs using Ad-vectors (p<0.01, Figure 5A and B).

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