Journal: ImmunoTargets and Therapy

Article Title: Characterizing CD8+ TEMRA Cells in CP/CPPS Patients: Insights from Targeted Single-Cell Transcriptomic and Functional Investigations

doi: 10.2147/ITT.S451199

Figure Lengend Snippet: Comparative analysis of CD8+ TEMRA cells derived from patients with CP-LS and HCs. PBMCs from patients with CP-LS and HCs were stimulated for 16 hours and analyzed via flow cytometry for the expression of the cytokines IFN-γ, TNF-α, granzyme B, and perforin. ( A ) Violin plot displaying the comparison of the expression of selected tropic chemokines and classical cytotoxicity activation genes and downregulated exhausted T-cell effector molecules in CD8+ TEMRA cells derived from the CP/CPPS group and HC group. ( B ) Representative flow cytometry dot plots of CD45RA and CCR7 expression were generated to identify four CD8+ T-cell subsets among the PBMCs of patients with CP-LS (right plot) and HCs (left plot). ( C ) Percentages of the CD45RA+CCR7+ (T-cell naïve), CD45RA-CCR7+ (TCM), CD45RA-CCR7- (TEM) and CD45RA+CCR7- (TEMRA) subsets within the CD8+T-cell population in PBMCs of patients with CP-LS (n=16) and HCs (n=16). ( D ) Percentages of IFN-γ+, TNF-α+, Granzyme B+, and Perforin+ CD8+ TEMRA cells from patients with CP-LS (n=16) and HCs (n=16). Values of *p<0.05, **p<0.01, and ***p<0.001 were considered significant.

Article Snippet: The cells were fixed with 100 μL of fixation reagent A (Fix/Perm medium A; Thermo Fisher Scientific) for 15 min. After washing, the cells were resuspended in 100 μL of permeabilization reagent B (Fix/Perm medium B; Thermo Fisher Scientific) and labeled with BV421-conjugated anti-perforin (BD Horizon, 563,393), BV510-conjugated anti-Granzyme B (BD Horizon, 563,388), BV786-conjugated anti-IFN-γ (BD Horizon, 563,731), and PE-conjugated anti-TNF-α (BD Horizon, 502,909) for 30 min at room temperature in the dark.

Techniques: Derivative Assay, Flow Cytometry, Expressing, Comparison, Activation Assay, Generated