Intestinal and mesenteric lymph node immune cell populations were characterised by flow cytometry. For intracellular cytokine staining, cells were incubated in T cell media (RPMI, 10% FBS, 1 mM sodium pyruvate, 2mM GLUTamax, 1X non-essential amino acids, 0.1 mM 2-β-mercaptoethanol, 10 mM HEPES, 1%Penicillin/Streptomycin) + 1X Cell Stimulation Cocktail with Protein Inhibitors (eBioscience) for 3 hours at 37°C before staining. Non-viable cells were stained using Live/Dead Fixable Aqua. Cells were permeabilised with CytoFix/CytoPerm (BD) followed by intracellular staining in PermBuffer (eBioscience).
All antibodies were purchased from eBioscience unless otherwise indicated. Antibodies used were CD45-SB600, TCRb-APC-Cy7, MHCII-FITC, CD4-PE-Cy7, CD8a-AF700, CD8b-PE, IFNg-PerCP-Cy5.5, TNFa-APC, IL-17A-bv421 (BioLegend), CD3e-FITC, TCRg-FITC, B220-FITC, MHCII-APC-e780, CD11b-PerCP-Cy5.5, CD11c-AF700, CD64-APC (BioLegend), SiglecF-PE (BD), Ly6G-Pe-Cy7 (BD) and F4/80-e450. Sample data were acquired with an Attune NxT flow cytometer coupled with an Attune CytKick Max autosampler. Data were analysed using FlowJo v10. Antibody details and concentrations are included in Supplementary Table 1.
All antibodies were purchased from eBioscience unless otherwise indicated. Antibodies used were CD45-SB600, TCRb-APC-Cy7, MHCII-FITC, CD4-PE-Cy7, CD8a-AF700, CD8b-PE, IFNg-PerCP-Cy5.5, TNFa-APC, IL-17A-bv421 (BioLegend), CD3e-FITC, TCRg-FITC, B220-FITC, MHCII-APC-e780, CD11b-PerCP-Cy5.5, CD11c-AF700, CD64-APC (BioLegend), SiglecF-PE (BD), Ly6G-Pe-Cy7 (BD) and F4/80-e450. Sample data were acquired with an Attune NxT flow cytometer coupled with an Attune CytKick Max autosampler. Data were analysed using FlowJo v10. Antibody details and concentrations are included in Supplementary Table 1.