Subfractionated CD62L^high and CD62L^low early effector cells possess similar properties in vitro. (A and B) Phenotypic characterization of CD62L^high and CD62L^low subsets. Five days after the initial stimulation with antigen and IL-2, CD62L^high and CD62L^low populations were sorted by MACS CD62L-positive selection column and analyzed by flow cytometry on day 6 for the expression of IL-7Rα; lymphoid-homing molecules β7 integrin and CCR7; the costimulatory molecule CD27; activation/effector markers CD44, CD69, CD25, and granzyme-B; and the senescence marker KLRG-1. MFI values after gating on CD8⁺Vβ13⁺ cells are shown. The MFI was determined for all groups at the same time under the same cytoflourometric settings using the same reagents and isotype-matched controls. Data shown are representative of 2 independent experiments. (C) CD62L^high and CD62L^low subsets have similar functional qualities. ⁵¹Cr, IFN-γ, and IL-2 release against hgp100_25–33–pulsed MCA-205 targets. MCA-205 cells plus influenza nucleoprotein peptide (np) were used as control. Where error bars are not visible, it is because they are obscured by the symbols used. Data shown are representative of 2 independent experiments. (D) CD62L^high and CD62L^low subsets have similar proliferative capacity. CD62L^high and CD62L^low cells were labeled with CFSE and were cultured with hgp100_25–33 peptide–pulsed on irradiated splenocytes in the presence of 30 IU/ml of rhIL-2. Cells were harvested from day 1 to day 4, and their CFSE content was determined. Results after gating on CD8⁺Vβ13⁺ population are shown. This experiment was performed twice, with similar results.