Limits...
Simian virus 40 vectors for pulmonary gene therapy.

Eid L, Bromberg Z, El-Latif MA, Zeira E, Oppenheim A, Weiss YG - Respir. Res. (2007)

Bottom Line: Moreover, our results showed vector presence in type II alveolar cells.The vector did not induce significant cellular immune response.These vectors appear to be capable of in vivo transduction of alveolar type II cells and may thus become a future therapeutic tool.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Anesthesiology and Critical Care Medicine, Hadassah - Hebrew University Medical Center, Jerusalem, 91120, Israel. luminita25@yahoo.com

ABSTRACT

Background: Sepsis remains the leading cause of death in critically ill patients. One of the primary organs affected by sepsis is the lung, presenting as the Acute Respiratory Distress Syndrome (ARDS). Organ damage in sepsis involves an alteration in gene expression, making gene transfer a potential therapeutic modality. This work examines the feasibility of applying simian virus 40 (SV40) vectors for pulmonary gene therapy.

Methods: Sepsis-induced ARDS was established by cecal ligation double puncture (2CLP). SV40 vectors carrying the luciferase reporter gene (SV/luc) were administered intratracheally immediately after sepsis induction. Sham operated (SO) as well as 2CLP rats given intratracheal PBS or adenovirus expressing luciferase served as controls. Luc transduction was evaluated by in vivo light detection, immunoassay and luciferase mRNA detection by RT-PCR in tissue harvested from septic rats. Vector abundance and distribution into alveolar cells was evaluated using immunostaining for the SV40 VP1 capsid protein as well as by double staining for VP1 and for the surfactant protein C (proSP-C). Immunostaining for T-lymphocytes was used to evaluate the cellular immune response induced by the vector.

Results: Luc expression measured by in vivo light detection correlated with immunoassay from lung tissue harvested from the same rats. Moreover, our results showed vector presence in type II alveolar cells. The vector did not induce significant cellular immune response.

Conclusion: In the present study we have demonstrated efficient uptake and expression of an SV40 vector in the lungs of animals with sepsis-induced ARDS. These vectors appear to be capable of in vivo transduction of alveolar type II cells and may thus become a future therapeutic tool.

Show MeSH

Related in: MedlinePlus

Lung pathology. H&E stained lung tissue, shown at ×20 and ×100 magnifications. Untreated control rats show normal lung histology; SO rats given intratracheal SV/luc also show normal histological appearance; 2CLP rats given intratracheal PBS or SV/luc show distinct ARDS pathology: alveoli filled with proteinaceous fluid, septal thickening, interstitial lymphocytic and neutrophilic infiltration.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2238754&req=5

Figure 1: Lung pathology. H&E stained lung tissue, shown at ×20 and ×100 magnifications. Untreated control rats show normal lung histology; SO rats given intratracheal SV/luc also show normal histological appearance; 2CLP rats given intratracheal PBS or SV/luc show distinct ARDS pathology: alveoli filled with proteinaceous fluid, septal thickening, interstitial lymphocytic and neutrophilic infiltration.

Mentions: Histological examination of the lung sections from 2CLP animals showed changes consistent with ARDS. Macroscopically: lungs were less aerated, covered with white fibrin patches and pleural fluid was found in different quantities. H&E stained sections depicted: alveoli filled with proteinaceous fluid, septal thickening, and interstitial neutrophilic infiltration (Figure 1). Mortality was present only in the septic (2CLP) animals. The 48 hrs mortality rate following 2CLP was similar to previously published numbers in sepsis induced ARDS [13-15].


Simian virus 40 vectors for pulmonary gene therapy.

Eid L, Bromberg Z, El-Latif MA, Zeira E, Oppenheim A, Weiss YG - Respir. Res. (2007)

Lung pathology. H&E stained lung tissue, shown at ×20 and ×100 magnifications. Untreated control rats show normal lung histology; SO rats given intratracheal SV/luc also show normal histological appearance; 2CLP rats given intratracheal PBS or SV/luc show distinct ARDS pathology: alveoli filled with proteinaceous fluid, septal thickening, interstitial lymphocytic and neutrophilic infiltration.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2238754&req=5

Figure 1: Lung pathology. H&E stained lung tissue, shown at ×20 and ×100 magnifications. Untreated control rats show normal lung histology; SO rats given intratracheal SV/luc also show normal histological appearance; 2CLP rats given intratracheal PBS or SV/luc show distinct ARDS pathology: alveoli filled with proteinaceous fluid, septal thickening, interstitial lymphocytic and neutrophilic infiltration.
Mentions: Histological examination of the lung sections from 2CLP animals showed changes consistent with ARDS. Macroscopically: lungs were less aerated, covered with white fibrin patches and pleural fluid was found in different quantities. H&E stained sections depicted: alveoli filled with proteinaceous fluid, septal thickening, and interstitial neutrophilic infiltration (Figure 1). Mortality was present only in the septic (2CLP) animals. The 48 hrs mortality rate following 2CLP was similar to previously published numbers in sepsis induced ARDS [13-15].

Bottom Line: Moreover, our results showed vector presence in type II alveolar cells.The vector did not induce significant cellular immune response.These vectors appear to be capable of in vivo transduction of alveolar type II cells and may thus become a future therapeutic tool.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Anesthesiology and Critical Care Medicine, Hadassah - Hebrew University Medical Center, Jerusalem, 91120, Israel. luminita25@yahoo.com

ABSTRACT

Background: Sepsis remains the leading cause of death in critically ill patients. One of the primary organs affected by sepsis is the lung, presenting as the Acute Respiratory Distress Syndrome (ARDS). Organ damage in sepsis involves an alteration in gene expression, making gene transfer a potential therapeutic modality. This work examines the feasibility of applying simian virus 40 (SV40) vectors for pulmonary gene therapy.

Methods: Sepsis-induced ARDS was established by cecal ligation double puncture (2CLP). SV40 vectors carrying the luciferase reporter gene (SV/luc) were administered intratracheally immediately after sepsis induction. Sham operated (SO) as well as 2CLP rats given intratracheal PBS or adenovirus expressing luciferase served as controls. Luc transduction was evaluated by in vivo light detection, immunoassay and luciferase mRNA detection by RT-PCR in tissue harvested from septic rats. Vector abundance and distribution into alveolar cells was evaluated using immunostaining for the SV40 VP1 capsid protein as well as by double staining for VP1 and for the surfactant protein C (proSP-C). Immunostaining for T-lymphocytes was used to evaluate the cellular immune response induced by the vector.

Results: Luc expression measured by in vivo light detection correlated with immunoassay from lung tissue harvested from the same rats. Moreover, our results showed vector presence in type II alveolar cells. The vector did not induce significant cellular immune response.

Conclusion: In the present study we have demonstrated efficient uptake and expression of an SV40 vector in the lungs of animals with sepsis-induced ARDS. These vectors appear to be capable of in vivo transduction of alveolar type II cells and may thus become a future therapeutic tool.

Show MeSH
Related in: MedlinePlus