Cd8a t cells isolation kit

On 6 dpi, splenocytes were isolated and CD8⁺ T cells were selected through negative selection with a CD8a⁺ T cells isolation kit (Miltenyi Biotec). 5‐μm polyester membrane Transwell inserts (Corning) were placed on a 24‐well plate (Corning) containing 600 μl of RPMI medium with 10% FBS and either no chemokines, 200 ng/ml of CXCL9, or 200 ng/ml of CXCL10. 1 × 10⁶ isolated CD8⁺ T cells were added to the upper chamber in 100 μl of medium lacking chemokines. The Transwell setup was incubated at 37°C for 3 h and activated CD3⁺CD8⁺LFA‐1⁺ or parasite‐specific CD3⁺CD8⁺Pb1⁺ T cells that crossed the membrane were quantified by flow cytometry. The chemotaxis index of each CD8⁺ T‐cell subset was determined by the ratio of cells recovered with chemokine‐containing medium to cells recovered with medium alone.