Flow Cytometry Analysis of Lung Immune Cells

BAL and mLN cells were processed and stained for flow cytometry as previously described [19] (link), [21] (link) with the following antibodies: CD16/32 (Fc blocker), CD4 (FITC), CD25 (PE-Cy7), and Foxp3 (APC) from eBioscience (San Diego, CA); CD3 (APC/Cy7) and CD8 (PerCP) from BioLegend (San Diego, CA). The MHC class I tetramer (PE) specific for H-2Db-resricted epitope of the influenza nucleoprotein (NP, D_bNP_366-74) of PR/8 was obtained from the NIH Tetramer Core Facility (Atlanta, GA). Stained cells were subsequently analyzed on a CyAn ADP Analyzer flow cytometer (Beckman Coulter, Inc., Fullerton, CA), and all flow cytometry data were analyzed via FlowJo Software (TreeStar, Ashland, OR). All flow cytometry analysis of T cells consisted of a doublet exclusion gate followed by gating of CD3⁺ cells for further analysis of CD4⁺ and CD8⁺ T cell populations.

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Milner J.J., Wang J., Sheridan P.A., Ebbels T., Beck M.A, & Saric J. (2014). 1H NMR-Based Profiling Reveals Differential Immune-Metabolic Networks during Influenza Virus Infection in Obese Mice. PLoS ONE, 9(5), e97238.

Publication 2014

Foxp3 (APC) CD3 (APC/Cy7) CD8 (PerCP) CyAn ADP Analyzer flow cytometer

Antibodies Cd25 Cd8 t cell Cells Epitope Fitc Flow cytometry H influenza Mhc class i Nucleoprotein Populations T cells Tetramer

Corresponding Organization :

Other organizations : University of North Carolina at Chapel Hill, Imperial College London

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Variable analysis

independent variables

Antibodies used for staining: CD16/32 (Fc blocker), CD4 (FITC), CD25 (PE-Cy7), Foxp3 (APC), CD3 (APC/Cy7), CD8 (PerCP)

dependent variables

Frequency and phenotype of CD4+ and CD8+ T cell populations

control variables

MHC class I tetramer (PE) specific for H-2Db-restricted epitope of the influenza nucleoprotein (NP, DbNP366-74) of PR/8

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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