Macs γδ t cell negative selection kit

RPMI-1640, FBS, non-essential amino acids, pyruvate, penicillin streptomycin, HEPES, 2-mercaptoethanol, and lymphoprep were from Thermo Fisher Scientific (Pittsburgh, PA). IL-2 and the MACS γδ T cell negative selection kit were from Miltenyi Biotec (Bergisch Gladbach, Germany). K562 cells were from Sigma-Aldrich (St. Louis, MO) while Research Blood Components (Boston, MA) supplied blood. The original passage number of the K562 cells is unknown, and K562 cells were maintained in suspension culture for no more than 3 months before thawing a new vial. Human interferon-γ ELISA MAX deluxe kit was from Biolegend (San Diego, CA).

RPMI-1640, FBS, non-essential amino acids, pyruvate, penicillin streptomycin, HEPES, 2-ME, HMBPP and FITC-γδ TCR antibody (clone 5A6.E91) were obtained through Fisher Scientific. IL-2 and the MACS γδ T cell negative selection kit were obtained through Miltenyi. PE-CD3 (UCHT1), APC-TNFα (MAb11), PE-IFNγ (4S.B3), FITC-IL-2 (MQ1-17H12), APC-CD45RA (HI-100), PerCP-eFluor710-CD27 (LG.7F9), FITC-Annexin V and PerCP-eFluor710-Annexin V were from eBioscience. K562 cells were from ATCC while Research Blood Components (Boston, MA) supplied blood. DiD, calcein AM, pcDNA3.1 and DNA primers were obtained through Life Technologies, while CRISPR/Cas9 repair templates were obtained through IDT. Restriction enzymes including XhoI, EcoRV and NheI were obtained through New England Biolabs. PE-BTN3A1 (CD277) antibody (BT3.1), FITC-CD183 (G025H7), functional grade anti-human TCR γ/δ Antibody (B1), and human IFNγ ELISA MAX deluxe kit were obtained from Biolegend while G418, PP2 and phytohaemagglutinin P were obtained from Sigma. Anti-myc (9E10) antibody was purified from hybridoma cells obtained from the Developmental Studies Hybridoma Bank at The University of Iowa. POM₂-C-HMBP was a kind gift of Dr. David Wiemer at the University of Iowa.