Journal: bioRxiv

Article Title: Metabolic and functional impairment of CD8 + T cells from the lungs of influenza-infected obese mice

doi: 10.1101/2020.04.19.047282

Figure Lengend Snippet: Lungs were harvested, digested, and homogenized into single cell suspension. Total CD8 + T cells from day 10-post influenza infected mice were isolated using negative selection magnetic bead separation. Cells were plated at 300,000 cells per well, and extracellular flux analysis was performed using the Agilent Seahorse XF e 96 flux analyzer. (A) Oxygen consumption rates (OCR), a surrogate of glycolysis, and (B) Extracellular Acidification Rates (ECAR), a surrogate of mitochondrial respiration, were measured in response to the Mitochondrial Stress Test for CD8 + T cells from Lean (n=11) and Obese (n=12) mice. (C) Basal OCR and (D) Basal ECAR were determined as the last point before oligomycin injection. Data represents mean ± SEM and were normalized to cell number. Wilcoxon rank sum test was used to compare groups. *p<0.05, **p<0.01.

Article Snippet: CD8 + T cells were isolated from the single cell suspension of mouse lungs at day 10 post PR8 infection using magnetic bead negative selection (Stemcell, Vancoover, Canada) in EasySep buffer (PBS + 2% FBS + 1mM EDTA).

Techniques: Suspension, Infection, Isolation, Selection, Injection

Journal: bioRxiv

Article Title: Metabolic and functional impairment of CD8 + T cells from the lungs of influenza-infected obese mice

doi: 10.1101/2020.04.19.047282

Figure Lengend Snippet: Lungs were harvested, digested, and homogenized into single cell suspension. Total CD8 + T cells from day 10-post influenza infected mice were isolated using negative selection magnetic bead separation. Cells were plated at 300,000 cells per well, and extracellular flux analysis was performed using the Agilent Seahorse XF e 96 flux analyzer and Mitochondrial Stress test for lean (n=11) and obese (n=12) mice. (A) Maximal respiration was measured as peak OCR following injection of 1.5μM FCCP. (B) Spare Respiratory Capacity was calculated as the difference in Max OCR minus Basal OCR. (C) ATP production was calculated as the difference between Basal OCR and the minimum OCR following injection of 0.5 μM oligomycin. (D) Glycolytic potential (OCR vs. ECAR) before and after injection of oligomycin. Data represents mean ± SEM and were normalized to cell number. Wilcoxon rank sum test was used to compare groups. *p<0.05.

Article Snippet: CD8 + T cells were isolated from the single cell suspension of mouse lungs at day 10 post PR8 infection using magnetic bead negative selection (Stemcell, Vancoover, Canada) in EasySep buffer (PBS + 2% FBS + 1mM EDTA).

Techniques: Suspension, Infection, Isolation, Selection, Injection