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Investigation of latent adenovirus 5 infection in guinea pig lung

University of British Columbia

The present study was based on the hypothesis that adenovirus E1A DNA persists in airway epithelial cells following a respiratory tract infection and is capable of amplifying cigarette smoke-induced airways inflammation. The experiments were conducted in a guinea pig model of latent adenovirus 5 (Ad5) infection where we have previously shown that latent infection increased the numbers of T-lymphocytes and macrophages in the lungs of animals acutely exposed to cigarette smoke. In the present study, we examined the lungs of 11 Ad5 infected and 12 sham infected animals 5 weeks post infection using PCR to demonstrate persistence of Ad5 E1A DNA, PCR in situ hybridization to localize this E1A DNA, and immunohistochemistry to detect the E1A protein expression in the lungs of Ad5 infected animals. Southern hybridization for the E1A PCR products demonstrated that Ad5 E1A DNA persisted in the lungs of 9 of 11 infected animals. PCR in situ hybridization localized this E1A DNA in the bronchiolar and alveolar epithelial cells in 5 of the 11 latently infected guinea pigs examined. Quantitative histology established that, on average, about 14 - 47 E1A positive cells were present in the total lung surface area (1.30 m² - 1.97 m²) of these lungs. Of the 11 latently infected animals, 2 were positive by immunohistochemistry for E1A protein expression in their lungs. None of the sham infected controls were positive in any of the above studies. These results show that latent adenoviral infection involves a small scattered population of airway epithelial cells in which viral DNA is easier to demonstrate than protein.

Thesis/Dissertation

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2009-06-12

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http://hdl.handle.net/2429/9052

Pathology

University of British Columbia

UBCV

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