Anti cd3ε percp

Anti–CD3ε-PerCP (100326), anti–CD8a-AF700 (100730), anti–CD8a-APC (100712), anti–PD-1-PE/DZ594 (109116), anti–Ki67-FITC (652410), anti–TCF1-PE (655208), anti–IFN-γ-BV421 (505830), anti–IFN-γ-PerCp/Cy5.5 (505822), anti–TNF-α-APC/Cy7 (506344), anti–TIM-3-PE/Cy7 (119716), anti–CD45-BV510 (103138), anti–Ly108 (Slamf6)-PE (134606), anti–PD-1-FITC (135214), anti–CD45.1-PE (110708), anti–CD45.2-AF488 (109816), purified anti-mouse CD3ε (100340), and purified anti-mouse CD28 (102116) were obtained from Biolegend. Anti–CD8-FITC (D271-4) and tetramer-SIINFEKL-APC (TS-5001-2c) were purchased from MBL. Cell Stimulation Cocktail Plus Protein Transport Inhibitors and FOXP3/TF Staining Buffer Set were bought from eBioscience. Naive CD8a⁺ T Cell Isolation and Tumor Dissociation Kits were obtained from Miltenyi Biotec.

Mice were infected as detailed in In vivo infections and footpad swelling section, and after 5 days of infection for MAYV or 6 days for CHIKV, the animals were sacrificed, and footpads were removed. Footpads homogenates were obtained after 2 hours incubation in collagenase VIII at 1mg/mL and passed thought a 70 μm cell strainer. Cells were counted and plated in 96-well U bottom. Cells were blocked with Fc Block (BD Biosciences) and then stained for flow cytometry analysis. The antibodies employed were anti-CD3ε-PerCP (BioLegend), anti-CD19-APC (BioLegend), anti-NK1.1-FITC (BioLegend), anti-CD45-APC (BD Biosciences), anti-CD11b-FITC (BioLegend), anti-Ly6G-PerCP (BioLegend) and anti-Ly6C-PE (BioLegend). Cells were also stained with a viability dye (Live/Dead fluorescent dye, Pacific Blue, Life Technologies). Samples were acquired on a FACS Verse flow cytometer (BD Biosciences) and analyzed with FlowJo software (Tree Star).