A novel method for intratracheal injection of infectious agents into mice

Abstract

Intratracheal injection is a traditional technique used in small animal studies of highly contagious airborne pathogens such as Mycobacterium tuberculosis. However, current techniques of intratracheal injection generally involve procedures that pose a risk of incident injury and infection for researchers, and may also cause collateral damage to experimental animals during the installation process. Here we describe an intratracheal injection method that was enabled by a three dimensional printing of a custom platform. This updated technique improved the overall ergonomics of intratracheal injection in mice, minimizing the risk of human injury and implementing the 3R (replacement, reduction and refinement) principle in mouse infection studies.

Extracellular vesicles deliver Mycobacterium RNA to promote host immunity and bacterial killing

“…M.tb input in the lung of mice was determined at day 1. Three weeks post‐infection, infected mice were treated by oral gavage with moxifloxacin at a dose of 50 mg/kg daily 6 days per week for 2 weeks, and one dose of EVs (5 μg/mouse, in 50 μl PBS) was administered intratracheally at 4 weeks post‐ M.tb infection as described previously . After the treatment, mouse serum was harvested via cardiac puncture and prepared using BD Microtainer Serum Separator Tube (BD), and cytokine production was measured by ELISA as described below.…”

Host cytosolic RNA sensing pathway promotes T Lymphocyte-mediated mycobacterial killing in macrophages

“…Mavs -/and wild-type C57BL/6 mice (8-10 weeks old, female) were intratracheally infected with M.avium 104 at a dose of 1X10 7 CFU as described previously [32]. M.avium input in the lung of mice was determined at 5 hr post-infection.…”

“…Infected AMs were washed with pre-cold PBS thrice and then intratracheally injected into mice at a dose of 2x10 5 cells/mouse in 15 μl PBS. At the time of AM injection, recipient mice had been infected with Gen S M.avium for 3 weeks as described [32]. At day 1, 3 and 6 post injection, mouse lungs were harvested, homogenized, and spread on 7H10 plus 10% OADC with or without gentamycin (200μg/ml).…”

Extracellular Vesicles promote host immunity during an M. tuberculosis infection through RNA Sensing

“…M.tb input in the lung of mice was determined at day 1. Three weeks post-infection, infected mice were treated by oral gavage with Moxifloxacin at a dose of 50 mg/kg daily six days per week for 2 weeks, and one dose of EVs (5 μg/mouse, in 50μl PBS) was administered intratracheally at 4 week post M.tb infection as described previously (Cheng et al, 2017). After the treatment, mouse serum was harvested via cardiac puncture and prepared using BD Microtainer Serum Separator Tube (BD), and cytokine production was measured by ELISA as described below.…”