Eight 10 week old female C57BL/6 mice were briefly anesthetized and bled using tail vein nick two days prior to i.n. infection with 104 or 105 FFU of DOG4 or mock infection and then after PBS or vaccine treatment (as described above) and on days 4, 6, 8 and 10 p.i. To control for natural diurnal fluctuations in levels of corticosterone, mice were bled between the hours of 9:00 am and 11:00 am each day. 25 µl serum from each animal was placed on ice and then spun at 2500× g at 4°C for 20 min. Plasma was stored at −80°C until assay. Corticosterone was measured by corticosterone EIA (Arbor Assays, Ann Arbor, MI) in duplicate according to manufacturer's instructions.
All experiments with mice were conducted in accordance with the Public Health Service Policy on Humane Care and Use of Laboratory Animals under protocols approved by the Institutional Animal Care and Use Committee of Thomas Jefferson University (Animal Welfare Assurance no. A3085-01).
All experiments with mice were conducted in accordance with the Public Health Service Policy on Humane Care and Use of Laboratory Animals under protocols approved by the Institutional Animal Care and Use Committee of Thomas Jefferson University (Animal Welfare Assurance no. A3085-01).