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. 2023 Jan 12;41(9):1296–1306. doi: 10.1038/s41587-022-01626-2

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a, Comparison of the cytotoxicities of NKCE molecules engaging only NKp46 (NKp46-Fc null-CD123) or coengaging NKp46 and CD16 (CD123-NKCE) against AML cell lines with and without CD32 and CD64 expression. THP-1 (CD32⁺ CD64⁺) cells and THP-1 subclones with inactivated CD32 (CD32-KO CD64⁺) or CD64 (CD32⁺ CD64-KO) expression were used as the targets, and purified resting NK cells from healthy donors were used as effectors. The data of one representative experiment among three are shown. b, Comparison of the cytotoxicities of CD123-IgG1⁺(black lozenge), NKp46-Fc null-CD123-NKCE (green circle) and CD123-NKCE (red square) against primary AML blasts expressing or not CD64. Primary AML blasts CD64-negative (AML no. 2) and CD64-positive (AML no. 4) were used as the targets and purified resting NK cells as effectors. The data for three healthy donor NK cells are shown. c, Cytotoxicity of CD123-NKCE against blasts from patients with AML. five blasts from patients with AML (AML nos. 1, 3, 5, 6 and 7) were used as targets, and purified NK cells from healthy donors (n = 9) were used as effectors. Results are shown for all the healthy NK cell donors tested. Data of a and c are presented as mean values ± s.d. d, Maximum cytotoxic activities of CD123-IgG1⁺ and CD123-NKCE molecules against blasts from patients with AML. Blast cells from seven patients with AML were used as targets and purified NK cells from ten healthy donors were used as effectors. Delta (Δ) maximum lysis, defined as percentage maximum lysis of the compound minus percentage background lysis of the isotype control molecule (IC-IgG1⁺ or IC-NKCE), were monitored from the dose response of each compound, and plotted separately for all couples of primary CD64-positive and CD64-negative AML sample/NK donor. (*P ≤ 0.05; **P ≤ 0.005; two-sided Wilcoxon matched-pairs signed rank test).