CD3+ and CD4+ T cells were freshly isolated from spleen and lymph nodes, some of which were shipped to Hamburg, Germany, on wet ice by negative selection using the EasySep Mouse T Cell Enrichment Kit or the EasySep Mouse CD4+ T Cell Enrichment Kit (STEMCELL Technologies Inc.). Cell purity (which was typically 95% T cells) was assessed by immunostaining with fluorescein isothiocyanate–conjugated anti-mouse TCRβ antibody (clone H57-597, BioLegend) and measured with a FACSCalibur flow cytometer (BD Biosciences).

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