Simian virus 40 vectors for pulmonary gene therapy.
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.


CONCLUSIONS
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.
Eid
Luminita
L
Department of Anesthesiology and Critical Care Medicine, Hadassah - Hebrew University Medical Center, Jerusalem, 91120, Israel. luminita25@yahoo.com
Bromberg
Zohar
Z
El-Latif
Mahmoud Abd
MA
Zeira
Evelyn
E
Oppenheim
Ariella
A
Weiss
Yoram G
YG
D000818:Animals; D036022:Capsid Proteins / Q000378:metabolism; D017930:Genes, Reporter / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics / Q000627:therapeutic use*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008168:Lung / Q000201:enzymology / Q000473:pathology; D011650:Pulmonary Alveoli / Q000201:enzymology / Q000473:pathology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012128:Respiratory Distress Syndrome / Q000382:microbiology / Q000628:therapy*; D018805:Sepsis / Q000150:complications; D013539:Simian virus 40 / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology; D014161:Transduction, Genetic
D036022:Capsid Proteins; D012333:RNA, Messenger; D008156:Luciferases