Cd3 percp cy5.5 clone 17a2

Cells were incubated 10 min with the TruStain fcX (clone 93) washed and incubated with the antibodies during 20 minutes followed by two washes with PBS. Monoclonal antibodies specific for mouse molecules were purchased from Biolegend: CD3-FITC (clone 17A2), CD3-APC (clone 17A2), CD3-PerCp/Cy5.5 (clone 17A2), CD8-Brillant Violet 421 (clone 53-6.7), CD45-PE (clone 30-F11), CD45-PErCP(clone 30-F11), CD45.1-PE/Cy7 (clone A20), CD45.1-FITC (clone A20), CD103-APC (clone 2E7), CD103-PerCP (clone 2E7), CD69-APC/Cy7 (clone H1.2F3), CD69-APC (clone H1.2F3), CD44-PerCP (clone IM7), IFN-γ-PE (clone XMG1.2), IFN-γ-APC (clone XMG1.2), TNF-α-APC/Cy7 (clone MP6-XT22), CD11b-FITC (clone M1/70), CD207-PE (clone 4C7), XCR1-APC (clone ZET), XCR1-PerCP-Cy5.5 (clone ZET), CD11c-PE/Cy7 (clone N418), MHCII-APC/Cy7 (clone M5/114.15.2), CD24-PerCP-Cy5.5 (clone M1/69), CD80-APC (clone 16-10A1), CD80-PE/Cy7 (clone 16-10A1), CD86 Brilliant Violet 421 (clone GL-1), CCR7-PE/Cy7 (clone 4B12), IL-2-PE/Cy7 (clone JES6-5H4) IL-12/23-APC (clone C15.6), granzyme B-APC (clone GB11) and viability dye Zombie Aqua (ref 423101). Samples were acquired in a FACSCanto II cytometer (BD Bioscience) and data were analyzed using FlowJo version X.0.7 (Tree Star, Inc.). Gating strategies for all flow cytometry experiments are shown in Supplementary Fig. 4.

Splenocyte populations of vaccinated mice were obtained at 7 days post-boost and were assessed by flow cytometry, as previously described, using the following directly conjugated Abs, CD3 PerCP-Cy5.5 (clone17A2; Biolegend, San Diego, CA, USA), CD4 APC/Cy7 (clone GK1.5; eBioscience, San Diego, CA, USA), CD8 APC (clone 53–6.7; eBioscience, San Diego, CA, USA), CD44 PE-Cy7 (clone IM7; eBioscience, San Diego, CA, USA), and CD62L Alexa Fluor 488 (Clone MEL-14; Biolegend, San Diego, CA, USA). Subsequently, cells were washed and fixed on ice with 2% paraformaldehyde in PBS and the number of CD4 and CD8 T effector cells was then quantified using an LSRII flow cytometer.