Influence of temperature on adenovirus-mediated gene transfer

Carlo Pellegrini, Timothy O'Brien, Anders Jeppsson, Lorraine A. Fitzpatrick, John Yap, Henry D. Tazelaar, Christopher G A McGregor

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Objective: The transfer of recombinant genes to donor organs may allow for novel therapeutic approaches to the challenges of acute and chronic rejection. Adenoviral vectors are capable of efficient gene transfer, but use of these vectors during donor organ preservation may be less efficient due to the low temperature. This study was designed to examine the effect of temperature on the efficiency of adenovirus-mediated gene transfer. Methods: Gene transfer to human endothelial cells, porcine vascular smooth muscle cells and cultured rat thoracic aortas was examined. Incubation with an adenoviral vector encoding for E. coli β-galactosidase was performed for I h at three different temperatures: 4°C, 10°C and 37°C. Transgene expression was assessed by histochemical staining for β-galactosidase in transduced cells and by evaluation of β-galactosidase activity in organ cultures. Results: Both in human endothelial cells and vascular smooth muscle cells the percentage of positively staining cells following transduction at 37°C was significantly greater than at 4°C and at 10°C (30.55 ± 7.26% vs. 14.29 ± 3.79% and 12.43 ± 2.47%, respectively for endothelial cells, P < 0.01 vs. 4°C and 10°C; and 28.25 ± 4.52% vs. 17.91 ± 3.76% and 16.63 ± 3.92%, respectively for smooth muscle cells, P < 0.05 vs. 4°C, P < 0.01 vs. 10°C). β-galactosidase activity was significantly greater in aortas transduced at 37°C than in vessels transduced at 4°C and 10°C (289 700 ± 113 300 vs. 149 600 °54 390 and 108 800 ± 23 140 relative chemiluminesce units/mg of total protein, respectively; P < 0.05 vs. 4°C, P < 0.001 vs. 10°C). Conclusions: The present study demonstrates that the efficiency of adenovirus-mediated gene transfer is significantly reduced at lower temperatures. The need for cold preservation of donor organs may render efficient adenovirus-mediated gent transfer more difficult in the transplantation setting.

Original languageEnglish (US)
Pages (from-to)599-603
Number of pages5
JournalEuropean Journal of Cardio-thoracic Surgery
Volume13
Issue number5
DOIs
StatePublished - May 1998

Fingerprint

Galactosidases
Adenoviridae
Temperature
Organ Preservation
Smooth Muscle Myocytes
Endothelial Cells
Genes
Tissue Donors
Vascular Smooth Muscle
Staining and Labeling
Organ Culture Techniques
Thoracic Aorta
Transgenes
Aorta
Swine
Transplantation
Escherichia coli
Proteins

Keywords

  • Adenovirus
  • Endothelial cells
  • Gene therapy
  • Gene transfer
  • Transplantation
  • Vascular smooth muscle cells

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Surgery

Cite this

Pellegrini, C., O'Brien, T., Jeppsson, A., Fitzpatrick, L. A., Yap, J., Tazelaar, H. D., & McGregor, C. G. A. (1998). Influence of temperature on adenovirus-mediated gene transfer. European Journal of Cardio-thoracic Surgery, 13(5), 599-603. https://doi.org/10.1016/S1010-7940(98)00064-5

Influence of temperature on adenovirus-mediated gene transfer. / Pellegrini, Carlo; O'Brien, Timothy; Jeppsson, Anders; Fitzpatrick, Lorraine A.; Yap, John; Tazelaar, Henry D.; McGregor, Christopher G A.

In: European Journal of Cardio-thoracic Surgery, Vol. 13, No. 5, 05.1998, p. 599-603.

Research output: Contribution to journalArticle

Pellegrini, C, O'Brien, T, Jeppsson, A, Fitzpatrick, LA, Yap, J, Tazelaar, HD & McGregor, CGA 1998, 'Influence of temperature on adenovirus-mediated gene transfer', European Journal of Cardio-thoracic Surgery, vol. 13, no. 5, pp. 599-603. https://doi.org/10.1016/S1010-7940(98)00064-5
Pellegrini C, O'Brien T, Jeppsson A, Fitzpatrick LA, Yap J, Tazelaar HD et al. Influence of temperature on adenovirus-mediated gene transfer. European Journal of Cardio-thoracic Surgery. 1998 May;13(5):599-603. https://doi.org/10.1016/S1010-7940(98)00064-5
Pellegrini, Carlo ; O'Brien, Timothy ; Jeppsson, Anders ; Fitzpatrick, Lorraine A. ; Yap, John ; Tazelaar, Henry D. ; McGregor, Christopher G A. / Influence of temperature on adenovirus-mediated gene transfer. In: European Journal of Cardio-thoracic Surgery. 1998 ; Vol. 13, No. 5. pp. 599-603.
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abstract = "Objective: The transfer of recombinant genes to donor organs may allow for novel therapeutic approaches to the challenges of acute and chronic rejection. Adenoviral vectors are capable of efficient gene transfer, but use of these vectors during donor organ preservation may be less efficient due to the low temperature. This study was designed to examine the effect of temperature on the efficiency of adenovirus-mediated gene transfer. Methods: Gene transfer to human endothelial cells, porcine vascular smooth muscle cells and cultured rat thoracic aortas was examined. Incubation with an adenoviral vector encoding for E. coli β-galactosidase was performed for I h at three different temperatures: 4°C, 10°C and 37°C. Transgene expression was assessed by histochemical staining for β-galactosidase in transduced cells and by evaluation of β-galactosidase activity in organ cultures. Results: Both in human endothelial cells and vascular smooth muscle cells the percentage of positively staining cells following transduction at 37°C was significantly greater than at 4°C and at 10°C (30.55 ± 7.26{\%} vs. 14.29 ± 3.79{\%} and 12.43 ± 2.47{\%}, respectively for endothelial cells, P < 0.01 vs. 4°C and 10°C; and 28.25 ± 4.52{\%} vs. 17.91 ± 3.76{\%} and 16.63 ± 3.92{\%}, respectively for smooth muscle cells, P < 0.05 vs. 4°C, P < 0.01 vs. 10°C). β-galactosidase activity was significantly greater in aortas transduced at 37°C than in vessels transduced at 4°C and 10°C (289 700 ± 113 300 vs. 149 600 °54 390 and 108 800 ± 23 140 relative chemiluminesce units/mg of total protein, respectively; P < 0.05 vs. 4°C, P < 0.001 vs. 10°C). Conclusions: The present study demonstrates that the efficiency of adenovirus-mediated gene transfer is significantly reduced at lower temperatures. The need for cold preservation of donor organs may render efficient adenovirus-mediated gent transfer more difficult in the transplantation setting.",
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AU - Pellegrini, Carlo

AU - O'Brien, Timothy

AU - Jeppsson, Anders

AU - Fitzpatrick, Lorraine A.

AU - Yap, John

AU - Tazelaar, Henry D.

AU - McGregor, Christopher G A

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N2 - Objective: The transfer of recombinant genes to donor organs may allow for novel therapeutic approaches to the challenges of acute and chronic rejection. Adenoviral vectors are capable of efficient gene transfer, but use of these vectors during donor organ preservation may be less efficient due to the low temperature. This study was designed to examine the effect of temperature on the efficiency of adenovirus-mediated gene transfer. Methods: Gene transfer to human endothelial cells, porcine vascular smooth muscle cells and cultured rat thoracic aortas was examined. Incubation with an adenoviral vector encoding for E. coli β-galactosidase was performed for I h at three different temperatures: 4°C, 10°C and 37°C. Transgene expression was assessed by histochemical staining for β-galactosidase in transduced cells and by evaluation of β-galactosidase activity in organ cultures. Results: Both in human endothelial cells and vascular smooth muscle cells the percentage of positively staining cells following transduction at 37°C was significantly greater than at 4°C and at 10°C (30.55 ± 7.26% vs. 14.29 ± 3.79% and 12.43 ± 2.47%, respectively for endothelial cells, P < 0.01 vs. 4°C and 10°C; and 28.25 ± 4.52% vs. 17.91 ± 3.76% and 16.63 ± 3.92%, respectively for smooth muscle cells, P < 0.05 vs. 4°C, P < 0.01 vs. 10°C). β-galactosidase activity was significantly greater in aortas transduced at 37°C than in vessels transduced at 4°C and 10°C (289 700 ± 113 300 vs. 149 600 °54 390 and 108 800 ± 23 140 relative chemiluminesce units/mg of total protein, respectively; P < 0.05 vs. 4°C, P < 0.001 vs. 10°C). Conclusions: The present study demonstrates that the efficiency of adenovirus-mediated gene transfer is significantly reduced at lower temperatures. The need for cold preservation of donor organs may render efficient adenovirus-mediated gent transfer more difficult in the transplantation setting.

AB - Objective: The transfer of recombinant genes to donor organs may allow for novel therapeutic approaches to the challenges of acute and chronic rejection. Adenoviral vectors are capable of efficient gene transfer, but use of these vectors during donor organ preservation may be less efficient due to the low temperature. This study was designed to examine the effect of temperature on the efficiency of adenovirus-mediated gene transfer. Methods: Gene transfer to human endothelial cells, porcine vascular smooth muscle cells and cultured rat thoracic aortas was examined. Incubation with an adenoviral vector encoding for E. coli β-galactosidase was performed for I h at three different temperatures: 4°C, 10°C and 37°C. Transgene expression was assessed by histochemical staining for β-galactosidase in transduced cells and by evaluation of β-galactosidase activity in organ cultures. Results: Both in human endothelial cells and vascular smooth muscle cells the percentage of positively staining cells following transduction at 37°C was significantly greater than at 4°C and at 10°C (30.55 ± 7.26% vs. 14.29 ± 3.79% and 12.43 ± 2.47%, respectively for endothelial cells, P < 0.01 vs. 4°C and 10°C; and 28.25 ± 4.52% vs. 17.91 ± 3.76% and 16.63 ± 3.92%, respectively for smooth muscle cells, P < 0.05 vs. 4°C, P < 0.01 vs. 10°C). β-galactosidase activity was significantly greater in aortas transduced at 37°C than in vessels transduced at 4°C and 10°C (289 700 ± 113 300 vs. 149 600 °54 390 and 108 800 ± 23 140 relative chemiluminesce units/mg of total protein, respectively; P < 0.05 vs. 4°C, P < 0.001 vs. 10°C). Conclusions: The present study demonstrates that the efficiency of adenovirus-mediated gene transfer is significantly reduced at lower temperatures. The need for cold preservation of donor organs may render efficient adenovirus-mediated gent transfer more difficult in the transplantation setting.

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KW - Endothelial cells

KW - Gene therapy

KW - Gene transfer

KW - Transplantation

KW - Vascular smooth muscle cells

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