Antibodies to cd16

Oral squamous carcinoma stem cells (OSCSCs) and oral squamous cell carcinoma (OSCCs) were isolated from patients with tongue tumors at UCLA (13 (link), 42 (link), 55 (link)). NK cells, OSCSCs, and OSCCs were cultured in RPMI 1640 (Invitrogen by Life Technologies, CA), supplemented with 10% fetal bovine serum (FBS) (Gemini Bio-Products, CA). Recombinant IL-2 was obtained from NIH-BRB. Antibodies to CD16 were purchased from Biolegend (San Diego, CA). Antibodies used for flow cytometry – IgG2, MHC-class I, and B7H1 (PD-L1) were purchased from Biolegend (San Diego, CA). MIA PaCA-2 (MP2), PL12, and Capan human pancreatic cancer cell lines were provided by Dr. Nicholas Cacalano (UCLA, School of Medicine, CA, USA), and were cultured in DMEM supplemented with 10% FBS. Cisplatin and Paclitaxel were purchased from Ronald Reagan Pharmacy at UCLA. ELISA kits for IFN-γ were purchased from Biolegend (San Diego, CA), and multiplex analysis kit was purchased from Millipore (Billerica, MA). Propidium iodide (PI) and chromium-51 was purchased from PeproTech (Cranbury, NJ, USA) and Perken Elmer (Waltham, MA, USA), respectively. Chromium Single cell 3’ Reagent kit v3, Cat#1000075 was purchased from10X Genomics (Pleasanton, CA, USA).

Rabbit anti-cystatin F polyclonal antibody, recombinant cistatin F and mouse 1D10 anti-cathepsin H Mab was prepared by our group in Ljubljana as reported [45 (link), 50 (link)]. Rabbit N1 antibody against N-terminal of cystatin and Δ15N cystatin F were prepared at Dundee University, Dundee, UK, [54 (link)]. Goat anti-cathepsin C pAb was from R&D Systems (Minneapolis, MN), mouse anti-GAPDH mAb was from Invitrogen and rabbit anti-CRK-L pAb was from Sigma. The dye identifying the lysosomes, LysoTracker, was from Molecular Probes (Eugene, OR). Secondary labelled antibodies goat anti-rabbit Alexa Fluor 488 and donkey anti-goat Alexa Fluor 555 were from Molecular Probes (Eugene, OR). Antibodies to CD16 were purchased from Biolegend (San Diego, CA). FITC anti-human/mouse granzyme B antibody was purchased from Biolegend (San Diego, CA).