Vδ2 percp clone b6

PBMCs were isolated from HC samples and then incubated with the following antibodies: CD3-FITC (clone SK7), TCR γδ-PE (clone IP26), Vδ1- PE-Cy7 (clone TS8.2), Vδ2-PerCP (clone B6), PE-Cy7 isotype control (clone P3.6.2.8.1), PE isotype control (clone eBM2a), FITC isotype control (clone MOPC-21), and PerCP isotype control (clone MOPC-21) (Biolegend, San Diego, USA; BD, Biosciences, San Jose, USA; Abcam Cambridge, UK). PBMCs were stained with different combinations of anti-human TCR γδ, Vδ1, Vδ2, and CD3 antibodies at room temperature in a dark room for 20 min. A total of 30,000–50,000 CD3\+ T cells were collected with a Cytoflex S flow cytometer (Beckman Coulter, USA), and data were analyzed by FlowJo software (FlowJo LLC, USA).

Peripheral blood mononuclear cells (PBMCs) were isolated from AML patients and HIs and then incubated with the following antibodies: CD45-V450 (clone H130), CD3-Alexa Fluor^® 700 (clone SP34-2), TCR γδ-PE-Cy7 (clone B1), Vδ1-FITC (clone TS8.2), Vδ2-PerCP (clone B6), PD-1-PE (clone EH12.2H7), Foxp3-Alexa Fluor^® 647 (clone 150D), PE isotype control (clone MOPC-21), and Alexa Fluor^® 647 isotype control (clone 259D/C7) (BioLegend, SanDiego, USA; BD Biosciences, San Jose, USA; Abcam Cambridge, UK). First, cells obtained from fresh PB samples were stained with surface markers including CD45, CD3, TCR γδ, Vδ1, Vδ2, and PD-1 at 4°C in the dark for 30 minutes. For Foxp3 expression detection, anti-Foxp3 antibody and Foxp3 Fix/Perm buffer were used according to the manufacturer’s instructions. The gating standards for PD-1 and Foxp3 were set using the isotype controls recommended by the manufacturer. A total of 30,000-50,000 CD3⁺ cells were collected with a BD FACS VERSE flow cytometer (BD Biosciences, San Jose, USA), and data were analyzed by Flowjo software (Flowjo LLC, USA) (38 (link)).