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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.

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Luciferase immunostaining. Was performed on the lung tissue harvested from 2CLP and SO rats given intratracheal SV/luc. Positive immunostaining appears as brown intracytoplasmatic coloration (Black arrows). Shown at ×100 magnification.
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Figure 3: Luciferase immunostaining. Was performed on the lung tissue harvested from 2CLP and SO rats given intratracheal SV/luc. Positive immunostaining appears as brown intracytoplasmatic coloration (Black arrows). Shown at ×100 magnification.

Mentions: Luc immunostaining was nonspecific, involving both alveolar type I type II cells. It was moderate in septic animals and low in SO animals (Figure 3). This may be due to moderate infectivity of the SV40 vector in the lungs or weak activity of the SV40 promoter in alveolar cells. This finding is also consistent with the literature, describing less transgene expression of some non – mammalian, non – vertebrate encoded proteins (e.g. luciferase, GFP, lac Z) commonly used as markers for transduction in SV40 vectors, than is usually seen with other vector systems (e.g., adenovirus) [18].


Simian virus 40 vectors for pulmonary gene therapy.

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

Luciferase immunostaining. Was performed on the lung tissue harvested from 2CLP and SO rats given intratracheal SV/luc. Positive immunostaining appears as brown intracytoplasmatic coloration (Black arrows). Shown at ×100 magnification.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: Luciferase immunostaining. Was performed on the lung tissue harvested from 2CLP and SO rats given intratracheal SV/luc. Positive immunostaining appears as brown intracytoplasmatic coloration (Black arrows). Shown at ×100 magnification.
Mentions: Luc immunostaining was nonspecific, involving both alveolar type I type II cells. It was moderate in septic animals and low in SO animals (Figure 3). This may be due to moderate infectivity of the SV40 vector in the lungs or weak activity of the SV40 promoter in alveolar cells. This finding is also consistent with the literature, describing less transgene expression of some non – mammalian, non – vertebrate encoded proteins (e.g. luciferase, GFP, lac Z) commonly used as markers for transduction in SV40 vectors, than is usually seen with other vector systems (e.g., adenovirus) [18].

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