Percp anti cd8a

APC-anti-CD3e, PE-Cy7-anti-CD3e, FITC-anti-CD3e, PE-anti-CD8a, PerCp-anti-CD8a, Alex700-anti-CD19, FITC-anti-CD44, APC-anti-CD44, V450-anti-CD44, PE-anti-IFN-γ, PE-Cy7-anti-TNF-α were purchased from BD Biosciences (Australia); PE-Cy7-anti-CD25, FITC-anti-CD27, APC-anti-CD27, Alexa647-anti-CD107a, eFluo450-anti-CD122, PE-anti-T-bet and Alexa700-anti-Eomes and purified anti-mouse MHC I-SIINFEKL complex (clone 25D) were purchased from eBioscience (Jomar Bioscience, Kensington, SA., Australia); PerCp-Cy5.5-anti-CD62L, PE-Cy7-anti-CD69 and purified anti-CD11c were purchased from BioLegend, (Australian Biosearch, Karrinyup, WA., Australia); APC-H-2Kb-SIINFEKL pentamer was purchased from ProImmune (Oxford, UK).

Single lung cell suspensions were prepared and stained as previously described.18 (link) Antibodies used were FITC-anti-TCRβ chain (BD, cat. no. 553171, clone H57-597), PE-anti-CD4 (BD, cat. no. 553652, clone H129.19), PerCP-anti-CD8a (BD, cat. no. 561092, clone 53-6.7), PerCP-Cy5.5-anti-CD11b (BD, cat. no. 561092, clone M1/70), FITC-anti-CD11c (BD, cat. no. 553801, clone HL3), PE-anti-MHCII (eBioscience, cat. no. 12-5321, clone M5/114.15.2), all at 1:15 dilution. Positive cells were identified using a FACSCanto (BD) by the following criteria: mDCs—CD11b⁺ CD11c⁺ MHCII⁺; T cells—TCRβ chain⁺ with CD4⁺ or CD8a⁺. Data analysed with BD FACsdiva.