Flow Cytometric Analysis
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For flow cytometric analysis of surface markers and intracellular cytokines, enriched C4+ T cells, and single-cell suspensions of lungs were stained with optimal concentrations of the following specific antibodies against surface molecules and intracellular cytokines: C8 α-V450, C4-V500, C11c-FITC, C62L-APC, Gr-1-APC, IFNγ-APC from , , , , and MHCII(I-A/I-E)-eFluor780 from eBioscience, C44-FITC, and IFNγ-PE fromand IL-22-PE from& .
- Fluorescence intensity was measured by using a FacsCantoII® flow cytometer equipped with a 405 nm, 488 nm and 633 nm laser.
- Analysis was performed utilizing the FCSExpress4 program (DeNovo™ Software) gated on leukocytes identified by the forward-scatter/side-scatter profile and further characterization as specified in the figures.
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