2.10. Immunophenotype and Cytotoxic Functional Analysis
Flasks of K562 cells were removed from the cell incubator and transferred into a conical tube. Cells were counted using a Countess 3 Automated Cell Counter (Invitrogen, Waltham, MA, USA) and spun down at 400× Analysis of acquired raw data was performed using FlowJo software (version 10, TreeStar, Ashland, OR, USA). Cell cultures containing only PBMC populations were used for immunophenotyping. Cell populations were determined as being CD3Data were processed as normalized to the control K562-only culture for each blood sample. Median fluorescence intensities of 1:1, 5:1, 10:1, and 20:1 ratios were divided by the K562-only control median fluorescence intensity, then multiplied by 100 to normalize data to the control cell viability of K562 only and expressed as percent cell viability as previously shown [