αβtcr pe

Flow cytometry analysis of immunomagnetically isolated γδ T cells was performed to ascertain their purity. Briefly, a fraction of isolated γδ T cells were washed with 1× phosphate-buffered saline (PBS), cold fixed with 1% paraformaldehyde for 15 min at 4°C and stained for cell surface markers using the following conjugated antibodies procured from BD Biosciences: Vδ2 TCR-FITC (clone B6), CD56-APC R-700 (clone NCAM16.2), αβ TCR-PE (clone T10B9.1A-31), CD14-PE (clone M5E2), CD11b PE-CF594 (clone ICRF44), CD19-BV786 (clone SJ25C1), Vδ2 TCR-PE (clone B6), and Vδ1 TCR-PerCP-Vio 700 (clone REA173). The cells were incubated for 30 min at 4°C and washed with FACS buffer. The purity and distribution of Vδ2 and Vδ1 populations in isolated γδ T cells were confirmed by flow cytometry using FACS Aria III flow cytometer (BD Biosciences, USA) and analysis performed using FlowJo software (TreeStar, Ashland, USA). The purity of γδ T cells isolated from peripheral blood of three healthy individuals was 96.4 ± 1.07%. As the distribution of Vδ2 population was higher (88.2 ± 5.92%) in the isolated γδ T cells as compared to Vδ1 (7.03 ± 1.26%), these are henceforth referred to as Vγ9Vδ2 T cells (Supplementary Figure S1).