Distinct CD16a features on human NK cells observed by flow cytometry correlate with increased ADCC.
Distinct CD16a features on human NK cells observed by flow cytometry correlate with increased ADCC.
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通过流式细胞术观察到的人类 NK 细胞的独特 CD16a 特征与 ADCC 增加相关。
DOI：
10.1038/s41598-024-58541-6
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发表时间：
2024
影响因子：
4.6
通讯作者：
Barb,AdamW
中科院分区：
文献类型：
--
作者：
Benavente,MariaCRodriguez;Hakeem,ZainabA;Davis,AlexanderR;Murray,NathanB;Azadi,Parastoo;Mace,EmilyM;Barb,AdamW
自然杀伤(NK)细胞破坏被抗体调理的组织。产生或鉴定具有更高效力的细胞的策略预计将加强基于NK细胞的免疫疗法。我们以前用Kifunensine治疗后产生的NK细胞具有更高的抗体依赖细胞介导的细胞毒(ADCC)，Kifunensine是一种针对甘露糖苷酶的抑制剂，在当时的糖蛋白加工途径早期。Kifunensine治疗还可提高Fcγ受体IIIa/CD16A的抗体结合亲和力。在这里，我们证明了抑制NK细胞N-糖链的处理增加了ADCC。我们通过CRIPSR-Cas9敲除另一种早期N-糖链处理酶MGAT1来减少N-糖链的处理，并表明这些细胞同样增加了抗体结合亲和力和ADCC。这些实验导致观察到，N-聚糖处理能力减弱的NK细胞在使用B73.1和3G8抗体结合两个不同的CD16A表位的流式细胞术实验中也显示出明显的表型。我们使用原代NK细胞对这种“亲和力分析”方法进行了评估，并通过与ADCC增加相关的流式细胞术鉴定了一个明显的变化和分化的群体。
Natural killer (NK) cells destroy tissue that have been opsonized with antibodies. Strategies to generate or identify cells with increased potency are expected to enhance NK cell-based immunotherapies. We previously generated NK cells with increased antibody-dependent cell mediated cytotoxicity (ADCC) following treatment with kifunensine, an inhibitor targeting mannosidases early in theN-glycan processing pathway. Kifunensine treatment also increased the antibody-binding affinity of Fc γ receptor IIIa/CD16a. Here we demonstrate that inhibiting NK cellN-glycan processing increased ADCC. We reducedN-glycan processing with the CRIPSR-CAS9 knockdown of MGAT1, another early-stageN-glycan processing enzyme, and showed that these cells likewise increased antibody binding affinity and ADCC. These experiments led to the observation that NK cells with diminishedN-glycan processing capability also revealed a clear phenotype in flow cytometry experiments using the B73.1 and 3G8 antibodies binding two distinct CD16a epitopes. We evaluated this “affinity profiling” approach using primary NK cells and identified a distinct shift and differentiated populations by flow cytometry that correlated with increased ADCC.