All animals used in this study were female BALB/c mice (Janvier Labs, France) housed in a minimal disease unit at Oslo University Hospital. All experiments were approved by the Norwegian Animal Research Authority.
Mice were anaesthetized by an intraperitoneal (i.p.) injection of ZRF (zolazepam [3.3 mg/mL], tiletamine [3.3 mg/mL], xylazine [0.45 mg/mL], fentanyl [2.6 μg/mL]) at 10 μL/g body weight. Immunizations were performed by first shaving the hind legs of anaesthetized mice, followed by an intramuscular (i.m.) injection of 50 μL DNA (0.5 mg/mL) solution into each quadriceps, immediately followed by five-pulse electroporation of the injection site with an AgilePulse system (Harvard Apparatus BTX, Holliston, MA). Protein vaccination was performed under anesthesia by i.m. injection of 50 μL vaccine solution in each quadriceps. For commercial vaccines, 2 × 50 μL of Flublok (Sanofi Pasteur, France) or a 1:1 mixture of Pandemrix:AS03 (GSK, Belgium) was delivered i.m. Viral influenza challenges were done by infecting anaesthetized mice intranasally (i.n.) with a virus dose of 5× the 50% lethal dose (LD50) in 10 μL per nostril. The LD50 dose was established by the Reed and Muench method and titrations of virus in mice. The viral strains used in this study were A/Puerto Rico/8/1934(H1N1), A/California/07/2009(H1N1), and RG14 [reassorted PR8 virus with H5 from A/Viet Nam/04/2005(H5N1)].
Mice were anaesthetized by an intraperitoneal (i.p.) injection of ZRF (zolazepam [3.3 mg/mL], tiletamine [3.3 mg/mL], xylazine [0.45 mg/mL], fentanyl [2.6 μg/mL]) at 10 μL/g body weight. Immunizations were performed by first shaving the hind legs of anaesthetized mice, followed by an intramuscular (i.m.) injection of 50 μL DNA (0.5 mg/mL) solution into each quadriceps, immediately followed by five-pulse electroporation of the injection site with an AgilePulse system (Harvard Apparatus BTX, Holliston, MA). Protein vaccination was performed under anesthesia by i.m. injection of 50 μL vaccine solution in each quadriceps. For commercial vaccines, 2 × 50 μL of Flublok (Sanofi Pasteur, France) or a 1:1 mixture of Pandemrix:AS03 (GSK, Belgium) was delivered i.m. Viral influenza challenges were done by infecting anaesthetized mice intranasally (i.n.) with a virus dose of 5× the 50% lethal dose (LD50) in 10 μL per nostril. The LD50 dose was established by the Reed and Muench method and titrations of virus in mice. The viral strains used in this study were A/Puerto Rico/8/1934(H1N1), A/California/07/2009(H1N1), and RG14 [reassorted PR8 virus with H5 from A/Viet Nam/04/2005(H5N1)].
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