Adaptive single-KIR+NKG2C+ NK cells expanded from superdonors show potent missing-self reactivity and efficiently control HLA-mismatched acute myeloid leukemia

Adoptive natural killer (NK) cell therapy in combination with hematopoietic stem cell transplantation (HSCT) can provide cure for acute myeloid leukemia (AML) but is in part dependent on variable NK cell reactivity. Adaptive NK cells are a highly potent NK cell subsets that can be utilized to maximize ‘missing-self’ reactivity against tumor cells. What this study adds: It describes a novel GMP-compliant protocol to expand clinically relevant numbers of adaptive NK cells from third-party ‘superdonors’. These NK cells provide strong reactivity in a mouse model of AML as well as against primary AML blasts ex vivo. How this study might affect research, practice or policy: These pre-clinical data demonstrate the feasibility of an NK cell-based cell therapy with a non-engineered and yet highly specific NK cell population, representing the first route to clinical testing of missing-self recognition. CITEseq was performed as outlined in Biolegend ‘TotalSeqTM-A Antibodies and Cell Hashing with 10x Single Cell 3' Reagent Kit v3 3.1 Protocol’ with minor modifications, using Biolegend oligo-conjugated antibodies and streptavidin TotalSeq reagents. Briefly, ADAPT-NK cells were stained with CD56-biotin mAb (Miltenyi, clone REA196), followed by TotalSeq antibodies and streptavidin-PE and Live/Dead Aqua (Invitrogen). Cells were subsequently sorted for viable CD56+ cells by flow cytometry. Submitter states: Raw data uploaded to EGA with accession EGAS00001006614<<<

过继性自然杀伤（natural killer, NK）细胞疗法联合造血干细胞移植（hematopoietic stem cell transplantation, HSCT）可治愈急性髓系白血病（acute myeloid leukemia, AML），但其疗效一定程度上依赖于可变的NK细胞反应性。适应性NK细胞（adaptive NK cells）是一类高效能的NK细胞亚群，可用于最大化针对肿瘤细胞的“缺失自我（missing-self）”免疫反应性。
本研究的新增贡献：本研究描述了一种符合良好生产规范（Good Manufacturing Practice, GMP）的新型方案，可从第三方“超级供者（superdonors）”体外扩增获得符合临床应用规模的适应性NK细胞。这类NK细胞在AML小鼠模型以及原代AML母细胞的离体实验中均表现出强劲的抗肿瘤反应活性。
本研究对科研、临床实践及政策的影响：这些临床前数据证实了基于非基因工程化且高特异性NK细胞群体的NK细胞疗法的可行性，为“缺失自我”识别的临床测试提供了首个可行路径。
CITEseq实验按照Biolegend公司《TotalSeqTM-A抗体及基于10x单细胞3'试剂试剂盒v3 3.1的细胞哈希实验方案》进行，并做了小幅修改，实验使用了Biolegend的寡核苷酸偶联抗体及链霉亲和素标记的TotalSeq试剂。简言之，ADAPT-NK细胞先用CD56生物素化单克隆抗体（美天旎，克隆号REA196）进行染色，随后加入TotalSeq抗体、链霉亲和素-PE及死活染料Aqua（赛默飞世尔Invitrogen）。后续通过流式细胞术分选出存活的CD56阳性细胞。
提交者说明：原始数据已上传至欧洲基因组-表型档案库（European Genome-phenome Archive, EGA），登录编号为EGAS00001006614。