T Cell Isolation

Spleen cell or lymph node cell suspensions were prepared in ice-cold PBS using a tissue homogenizer. Erythrocytes were depleted by hypo-osmotic shock. Highly purified (>98%) CD3⁺ T cells were obtained using the Pan T Cell Isolation MACS^® kit from Miltenyi Biotec (Cambridge, MA, United States), as per the manufacturer’s instructions. Highly purified CD4⁺ or CD8⁺ T cells were isolated by negative selection from lymph node cell preparations using CD4⁺ or CD8⁺ T Cell MACS^® isolation kits, as per the manufacturer’s instructions.