Pneumocystis Infection Model in Mice

Mice were fed mouse chow containing trimethoprim-sulfamethoxazole for 3 weeks to clear possible previous infections (72 (link)) and then allowed 2 weeks to clear the medication (73 (link)). Throughout experimentation, mice were maintained on an antibiotic regimen (500 mg/liter) in drinking water, alternating monthly between cephalexin and amoxicillin. Mice were intranasally infected with 0.5 × 10⁶P. murina cysts (ATCC) intranasally on day 0 and day 7 and monitored for 20% weight loss and subjective moribund characteristics resulting from PCP. Before sacrifice, blood oxygen saturation was measured to indicate compromise of pulmonary function and active pneumonia via the MouseOx System (Starr Life Sciences, Oakmont, PA). Upon sacrifice via carbon dioxide asphyxiation, lungs were weighed to assess general fluid influx into lung parenchyma. Mice that did not reach a terminal endpoint were allowed to live a minimum of 150 days. Depletion of CD4 T cells was accomplished by intraperitoneal injection of 0.3 mg GK1.5 mAb (BioXcell, New Lebanon, NH; or PBS control) on days −4 and −1 before infection, and weekly thereafter. Depletion status was monitored by flow cytometry analysis of blood periodically throughout the experiment.

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Neier S.C., Ferrer A., Wilton K.M., Smith S.E., Kelcher A.M., Pavelko K.D., Canfield J.M., Davis T.R., Stiles R.J., Chen Z., McCluskey J., Burrows S.R., Rossjohn J., Hebrink D.M., Carmona E.M., Limper A.H., Kappes D.J., Wettstein P.J., Johnson A.J., Pease L.R., Daniels M.A., Neuhauser C., Gil D, & Schrum A.G. (2019). The early proximal αβ TCR signalosome specifies thymic selection outcome through a quantitative protein interaction network. Science immunology, 4(32), eaal2201.

Publication 2019

MouseOx System GK1.5 mAb

Amoxicillin Antibiotic Asphyxiation Blood analysis Blood oxygen saturation Carbon dioxide Cd4 t cells Cephalexin Cysts Flow cytometry Infection Intraperitoneal injection Lung Mab gk1 5 Medication Mice Pneumonia Regimen Trimethoprim sulfamethoxazole

Corresponding Organization :

Other organizations : Mayo Clinic, Mayo Clinic in Arizona, University of Missouri, Peter Doherty Institute, University of Melbourne, QIMR Berghofer Medical Research Institute, University of Queensland, Australian Regenerative Medicine Institute, ARC Centre of Excellence in Advanced Molecular Imaging, Cardiff University, Monash University, Fox Chase Cancer Center, University of Minnesota

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Variable analysis

independent variables

Depletion of CD4 T cells accomplished by intraperitoneal injection of 0.3 mg GK1.5 mAb (or PBS control) on days -4 and -1 before infection, and weekly thereafter

dependent variables

Weight loss (20% weight loss as a terminal endpoint)
Subjective moribund characteristics resulting from PCP
Blood oxygen saturation (to indicate compromise of pulmonary function and active pneumonia)
Lung weight (to assess general fluid influx into lung parenchyma)

control variables

Mice were maintained on an antibiotic regimen (500 mg/liter) in drinking water, alternating monthly between cephalexin and amoxicillin
Mice were fed mouse chow containing trimethoprim-sulfamethoxazole for 3 weeks to clear possible previous infections and then allowed 2 weeks to clear the medication

positive control

PBS control for CD4 T cell depletion

negative control

Not explicitly mentioned

Annotations

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