Cd123 clone 6h6

Lung cells from HSPC-engrafted MISTRG mice were isolated as described above. Cell surface staining with fluorescent antibodies was performed as previously described [17 (link), 18 (link)]. The following antibodies were used for flow cytometry: CD127 (clone A019D5, Biolegend), CRTH2 (clone BM16, Biolegend), CD94 (clone DX22, Biolegend), CXCR4 (clone 12G5, Biolegend), CXCR6 (clone K041E5, Biolegend), GITR (clone 108-17, Biolegend), CD16 (clone 3G8, Biolegend), CD69 (clone FN50, Biolegend), CD117 (104D2D1, Beckman Coulter), HLA-DR (clone G46-6, BD Biosciences), CD2 (clone RPA-2.10, BD Biosciences), CD56 (clone NCAM16.2, BD Biosciences), CD81 (clone JS-81, BD Biosciences), CD94 (clone HP-3D9, BD Biosciences), CD98 (clone UM7F8, BD Biosciences), CD103 (clone Ber-ACT8, BD Biosciences), and CD45RA (clone HI100, BD Biosciences). In all experiments, ILCs were defined as human CD45⁺Lin^-CD3⁻TCRαβ⁻CD127⁺CD94⁻ cells. Lineage markers included CD14 (clone M5E2, Biolegend), CD19 (clone HIB19, Biolegend), CD11c (clone Bu15, Biolegend), CD123 (clone 6H6, Biolegend), FcεRIα (clone AER-37 (CRA-1), Biolegend), CD34 (clone 581, BD Biosciences), TCRαβ (clone IP26) and CD3 (clone SK7).

Ex vivo expanded primary AML cells of 9 different patients were co-cultured with freshly isolated healthy donor NK cells, at an E:T ratio of 5:1 in an ex vivo long term culture system as described by Krupka and coworkers [29 (link), 54 (link)]. Antibodies were added at a final concentration of 10 nM. After 24 hours, cells were harvested, stained for CD16 (clone B73.1), CD56 (clone HCD 56), CD33 (clone WM53) and in concrete cases CD123 (clone 6H6; all antibodies from Biolegend) and analyzed by flow cytometry with a BD LSR II (Becton Dickinson, Heidelberg, Germany). The percentage of residual CD33 or CD123 positive cells in treated cultures relative to control cultures was used to determine licMAB-mediated cellular cytotoxicity. Patient characteristics are summarized in Supplementary Table S1.