Intratracheal Inflammation Modulation in Mice

This study was carried out in strict accordance with the recommendations in the Guide for the Care and use of Laboratory Animals of the National Institutes of Health. The protocol was approved by Texas A&M University Animal Use and Care Committee approved the protocol (Protocol Number: IACUC 2017–0414). 5-week old 20 g male and female C57BL/6 mice (Jackson Laboratories, Bar Harbor, ME) were given an oropharyngeal aspiration of 1 mg/kg (or less where indicated) ORN06/LyoVec (a ssRNA oligonucleotide with 6 UGU repeats complexed with a lipid which protects the ssRNA from degradation; henceforth referred to as ORN06) (InvivoGen, San Diego, CA) in 50 μl phosphate-buffered saline (PBS), or an oropharyngeal aspiration of an equal volume of PBS as a control, following [51 (link),52 (link)]. At 24 and 48 hours after ORN06 insult, the control mice and some of the ORN06-treated mice were given intraperitoneal injections of 100 μl PBS, or SAP diluted to a dose of 2.5 mg/kg in 100 μl PBS. All the mice were monitored twice daily to observe any sign of distress. At day 3, mice were euthanized by CO₂ inhalation, and bronchoalveolar lavage fluid (BALF) was obtained as previously described [34 (link),51 (link)]. All mice survived until euthanasia at day 3, and 20 mice were used in the study.