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from Charles River Laboratories (Montreal, Quebec, Saucony Grid 9000 Black Canada) and the Cancer Research Centre (Frederick, MD), respectively. Animals were allowed to acclimatize for 5 d before use. Animals receiving RSV were housed separately from sham inoculated animals in large polycarbonate cages.
of pageMETHODSAnimals. Female, juvenile CH and STR2 (1 mo old, body weight 250 g) were obtained Saucony Shadow Vegan Slate
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Otherwise, all animals were kept under similar conditions of alternating 12 h light dark cycles and free access to guinea pig chow (Ralston Purina Corp. , St. Louis, MO) and water. This study was approved by the Committee on Animal Care of the University of British Columbia, and all animals were kept in accordance with standards of the Canadian Council on Animal Care (27). Virus preparation and inoculation.
3 104 plaque forming units (pfu)] or sham vehicle (uninfected HEp 2 .
Human Long strain, type A RSV (American Type Culture Collection, Manassas, VA) was propagated on HEp 2 cell monolayers (28). Working stocks of RSV were prepared by collection of the supernatant from lysed RSV infected HEp 2 cells and stored at Preparations were free of Mycoplasma as determined by culture of aliquots (Microbiology Laboratory, St. Paul's Hospital, Vancouver, BC, Canada), and endotoxin levels were below the limits of detection of Limulus E Toxate assay (Sigma Chemical Co. ). Guinea pigs were anesthetized via inhalation of 3% halothane and received 300 of either RSV containing inoculum [5.
in and STR2 were used as the reference allergy resistant strain. We performed viral culture on lung homogenates to quantify titers of intrapulmonary, replicating RSV; semiquantitative reverse transcription polymerase chain reaction (RT PCR) to measure IFN and IL 5 mRNA levels in lung homogenates, with calculation of IFN mRNA ratios to assess host Th1/Th2 balance; inhalational acetylcholine (ACh) challenge to test for RSV associated nonspecific AHR; and quantitative histology for T cells and eosinophils (14,16) to measure the extent of airway inflammation Saucony All Black Shoes
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