Transbronchial gene transfer of endothelial nitric oxide synthase to transplanted lungs

Anders Jeppsson, Carlo Pellegrini, Timothy O'Brien, Virginia M. Miller, Henry D. Tazelaar, Christopher G A McGregor

Research output: Contribution to journalArticle

Abstract

Background. Experiments were designed to study the efficiency, distribution, and toxicity of transbronchial adenoviral-mediated transfer of endothelial constitutive nitric oxide synthase (ecNOS) gene to transplanted lungs. Methods. Syngeneic orthotopic single-lung transplantation in the rat was performed after airway administration (300 μL, 1 x 109 pfu/mL) of either the ecNOS gene or the marker gene β-Gal (control group) to donor lungs (n = 4 each). After 4 days, transgene expression, inflammation, and the presence of apoptosis in the transplanted lungs were assessed by molecular, immunohistochemical, and histologic techniques. Results. Gene transfer was confirmed by a positive polymerase chain reaction signal for the recombinant ecNOS gene, and recombinant messenger RNA by reverse transcription polymerase chain reaction. Positive immunohistochemical staining for ecNOS was present in more than 75% of pneumocytes only in ecNOS transduced lungs. Calcium- dependent nitric oxide synthase activity was increased in ecNOS- compared with βGal-transduced lungs (2,139 ± 756 versus 47 ± 28 pmol · mg protein-1 · h-1; p <0.05). Minimal to mild inflammation was observed in all transplanted lungs; fewer than 0.5% of cells in both groups were apoptotic. Conclusions. Transbronchial transfer of ecNOS gene to the transplanted lung results in transduction of pneumocytes with expression of a functionally active transgene product.

Original languageEnglish
Pages (from-to)318-324
Number of pages7
JournalAnnals of Thoracic Surgery
Volume66
Issue number2
DOIs
Publication statusPublished - Aug 1998

Fingerprint

Nitric Oxide Synthase Type III
Lung
Genes
Alveolar Epithelial Cells
Transgenes
Inflammation
Histological Techniques
Polymerase Chain Reaction
Lung Transplantation
Nitric Oxide Synthase
Reverse Transcription
Apoptosis
Staining and Labeling
Calcium
Control Groups
Messenger RNA
Proteins

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Surgery

Cite this

Transbronchial gene transfer of endothelial nitric oxide synthase to transplanted lungs. / Jeppsson, Anders; Pellegrini, Carlo; O'Brien, Timothy; Miller, Virginia M.; Tazelaar, Henry D.; McGregor, Christopher G A.

In: Annals of Thoracic Surgery, Vol. 66, No. 2, 08.1998, p. 318-324.

Research output: Contribution to journalArticle

Jeppsson, Anders ; Pellegrini, Carlo ; O'Brien, Timothy ; Miller, Virginia M. ; Tazelaar, Henry D. ; McGregor, Christopher G A. / Transbronchial gene transfer of endothelial nitric oxide synthase to transplanted lungs. In: Annals of Thoracic Surgery. 1998 ; Vol. 66, No. 2. pp. 318-324.
@article{fc0ba24291cf4c37bb124aa842b639ef,
title = "Transbronchial gene transfer of endothelial nitric oxide synthase to transplanted lungs",
abstract = "Background. Experiments were designed to study the efficiency, distribution, and toxicity of transbronchial adenoviral-mediated transfer of endothelial constitutive nitric oxide synthase (ecNOS) gene to transplanted lungs. Methods. Syngeneic orthotopic single-lung transplantation in the rat was performed after airway administration (300 μL, 1 x 109 pfu/mL) of either the ecNOS gene or the marker gene β-Gal (control group) to donor lungs (n = 4 each). After 4 days, transgene expression, inflammation, and the presence of apoptosis in the transplanted lungs were assessed by molecular, immunohistochemical, and histologic techniques. Results. Gene transfer was confirmed by a positive polymerase chain reaction signal for the recombinant ecNOS gene, and recombinant messenger RNA by reverse transcription polymerase chain reaction. Positive immunohistochemical staining for ecNOS was present in more than 75{\%} of pneumocytes only in ecNOS transduced lungs. Calcium- dependent nitric oxide synthase activity was increased in ecNOS- compared with βGal-transduced lungs (2,139 ± 756 versus 47 ± 28 pmol · mg protein-1 · h-1; p <0.05). Minimal to mild inflammation was observed in all transplanted lungs; fewer than 0.5{\%} of cells in both groups were apoptotic. Conclusions. Transbronchial transfer of ecNOS gene to the transplanted lung results in transduction of pneumocytes with expression of a functionally active transgene product.",
author = "Anders Jeppsson and Carlo Pellegrini and Timothy O'Brien and Miller, {Virginia M.} and Tazelaar, {Henry D.} and McGregor, {Christopher G A}",
year = "1998",
month = "8",
doi = "10.1016/S0003-4975(98)00552-9",
language = "English",
volume = "66",
pages = "318--324",
journal = "Annals of Thoracic Surgery",
issn = "0003-4975",
publisher = "The Society of Thoracic Surgeons. Published by Elsevier Inc",
number = "2",

}

TY - JOUR

T1 - Transbronchial gene transfer of endothelial nitric oxide synthase to transplanted lungs

AU - Jeppsson, Anders

AU - Pellegrini, Carlo

AU - O'Brien, Timothy

AU - Miller, Virginia M.

AU - Tazelaar, Henry D.

AU - McGregor, Christopher G A

PY - 1998/8

Y1 - 1998/8

N2 - Background. Experiments were designed to study the efficiency, distribution, and toxicity of transbronchial adenoviral-mediated transfer of endothelial constitutive nitric oxide synthase (ecNOS) gene to transplanted lungs. Methods. Syngeneic orthotopic single-lung transplantation in the rat was performed after airway administration (300 μL, 1 x 109 pfu/mL) of either the ecNOS gene or the marker gene β-Gal (control group) to donor lungs (n = 4 each). After 4 days, transgene expression, inflammation, and the presence of apoptosis in the transplanted lungs were assessed by molecular, immunohistochemical, and histologic techniques. Results. Gene transfer was confirmed by a positive polymerase chain reaction signal for the recombinant ecNOS gene, and recombinant messenger RNA by reverse transcription polymerase chain reaction. Positive immunohistochemical staining for ecNOS was present in more than 75% of pneumocytes only in ecNOS transduced lungs. Calcium- dependent nitric oxide synthase activity was increased in ecNOS- compared with βGal-transduced lungs (2,139 ± 756 versus 47 ± 28 pmol · mg protein-1 · h-1; p <0.05). Minimal to mild inflammation was observed in all transplanted lungs; fewer than 0.5% of cells in both groups were apoptotic. Conclusions. Transbronchial transfer of ecNOS gene to the transplanted lung results in transduction of pneumocytes with expression of a functionally active transgene product.

AB - Background. Experiments were designed to study the efficiency, distribution, and toxicity of transbronchial adenoviral-mediated transfer of endothelial constitutive nitric oxide synthase (ecNOS) gene to transplanted lungs. Methods. Syngeneic orthotopic single-lung transplantation in the rat was performed after airway administration (300 μL, 1 x 109 pfu/mL) of either the ecNOS gene or the marker gene β-Gal (control group) to donor lungs (n = 4 each). After 4 days, transgene expression, inflammation, and the presence of apoptosis in the transplanted lungs were assessed by molecular, immunohistochemical, and histologic techniques. Results. Gene transfer was confirmed by a positive polymerase chain reaction signal for the recombinant ecNOS gene, and recombinant messenger RNA by reverse transcription polymerase chain reaction. Positive immunohistochemical staining for ecNOS was present in more than 75% of pneumocytes only in ecNOS transduced lungs. Calcium- dependent nitric oxide synthase activity was increased in ecNOS- compared with βGal-transduced lungs (2,139 ± 756 versus 47 ± 28 pmol · mg protein-1 · h-1; p <0.05). Minimal to mild inflammation was observed in all transplanted lungs; fewer than 0.5% of cells in both groups were apoptotic. Conclusions. Transbronchial transfer of ecNOS gene to the transplanted lung results in transduction of pneumocytes with expression of a functionally active transgene product.

UR - http://www.scopus.com/inward/record.url?scp=0031711929&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031711929&partnerID=8YFLogxK

U2 - 10.1016/S0003-4975(98)00552-9

DO - 10.1016/S0003-4975(98)00552-9

M3 - Article

C2 - 9725363

AN - SCOPUS:0031711929

VL - 66

SP - 318

EP - 324

JO - Annals of Thoracic Surgery

JF - Annals of Thoracic Surgery

SN - 0003-4975

IS - 2

ER -