Pe anti human cd56

20-Deoxyingenol 3-angelate (DI3A) was purchased from BioBioPha (Yunnan, China). Ingenol 3-angelate (I3A) was obtained from AdipoGen (San Diego, CA, USA). Calcein-AM was supplied by Sigma-Aldrich (St. Louis, MO, USA). Recombinant human IL-2 protein was provided by PeproTech (Rehovot, Israel). PE anti-human CD56, FITC anti-human CD107α, APC anti-human IFN-γ, and murine isotype controls (IgG1κ-PE, IgG1κ-FITC and IgG1κ–APC) were purchased from BioLegend Inc. (San Diego, CA, USA). CellTiter-Glo Luminescent Cell Viability Assay Kit was obtained from Promega (Madison, WI, USA). Fixation/Permeabilization Solution Kit with BD GolgiStop™ was provided by BD Biosciences (CA, USA).

NK cells were collected and treated with MTB H37Rv protein lysates as described above. And infected monocytes were cocultured with NK cells (2 × 10⁵/well) at a ratio of 1 : 2 at 37°C in a humidified 5% CO₂ atmosphere. After 4 h cultivation, cells were collected and incubated with specific antibodies at manufacturer's recommended concentrations at 4°C for 30 min to analyze the percentage of nonviable CD14+ monocytes by flow cytometry. The antibodies used in the study were as follows: PE anti-human CD56, brilliant violet 510™ anti-human CD16, APC anti-human CD14, and brilliant violet 711™ anti-human CD3 (Biolegend, San Diego, CA, USA). And fixability viability stain 440UV (FVS440UV, BD Biosciences, USA) was used for discrimination of nonviable from viable CD14+ monocytes to measure NK cell cytotoxicity. The cells were detected using BD LSRFortessa™ Flow Cytometer (BD Biosciences, USA), and results were analyzed using the BD FACSDiva software (version 8.0.2).

To isolate pDCs, cDCs, and monocytes, human PBMCs were labeled with Alexa 647 anti-human CD11c (301620, clone 3.9, BioLegend), anti-human HLA-DR (Class III) PE-Texas conjugate (MHLDR17, Life Technology), BV421 anti-human CD14 (325627, clone HCD14, BioLegend), PE-Cy7 anti-human CD123 (560826, clone 7G3, BD Biosciences), APC-Fire 750 anti-human CD8a (301065, clone RPA-T8, BioLegend), APC-Fire 750 anti-human CD20 (302357, clone 2H7, BioLegend), APC-H7 anti-human CD3 (560275, clone SK7, BD Pharmingen) and Live/Dead Fixable Aqua Dead Cell Stain Kit (L34966, Invitrogen). Labeled cells were fixed with 4% PFA in PBS for 5 min, then cell marker expression was analyzed and cells were isolated with BD FACSAria II (BD Biosciences). The gating strategy is shown in Supplementary Figure S4.
To isolate CD4⁺ T, CD8⁺ T, NK, and B cells, human PBMCs were labeled with APC-H7 anti-human CD3 (560275, BD Bioscience), PE-Cy7 anti-human CD4 (317413, Biolegend), PE anti-human CD56 (355503, Biolegend), FITC anti-human CD8 (300905, Biolegend), APC anti-human CD19 (302211, Biolegend), and Live/Dead Fixable Aqua Dead Cell Stain Kit (L34966, Invitrogen). Labeled cells were fixed with 4% PFA in PBS for 5 min, then expression markers were analyzed and cells were isolated with BD FACSAria II (BD Biosciences). The gating strategy is shown in Supplementary Figure S4.