表达 FCGR3 两种等位基因变体组合的 NK-92 细胞系介导的 NK 细胞毒性。

IF 3 Q3 IMMUNOLOGY
Antibodies Pub Date : 2024-07-12 DOI:10.3390/antib13030055
Marta Freitas Monteiro, Maria Papaserafeim, Matteo Andreani, Aline Réal, Athanasios Kouklas, Daniela Reis Galvão, Jörg D Seebach, Gisella L Puga Yung
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引用次数: 0

摘要

自然杀伤(NK)细胞在监控病毒感染和癌症方面发挥着重要作用。NK细胞的抗体依赖性细胞毒性(ADCC)和直接细胞毒性分别是通过Fcγ受体IIIA(FcγRIIIa/CD16)和激活/抑制NK受体的配体识别抗体包被的靶细胞介导的。FCGR3A 基因的等位基因变异包括与单克隆抗体(mAb)疗法疗效相关的高亲和性单核苷酸多态性(SNP)rs396991(V176F)和 SNP rs10127939(L66H/R)。FCGR3A SNPs 对 NK 细胞效应功能的贡献仍存在争议;因此,我们生成了表达这些 SNPs 特定组合的八种 NK-92 细胞系,并测试了它们的细胞毒性。用含有不同 FCGR3A SNPs 组合的质粒稳定转染 NK-92 细胞。分别使用新一代测序技术(NGS)和流式细胞术检测信使 RNA 和 FcγRIIIa/CD16 细胞表面表达。所有经 FcγRIIIa/CD16 转染的 NK-92 细胞系都对三种不同的靶细胞系表现出了强大的 ADCC 能力,但差别不大。此外,还观察到对 K562 靶细胞的直接 NK 细胞毒性增强,这表明 FcγRIIIa/CD16 在直接 NK 细胞毒性中的机制作用。总之,我们生成了八种经 FcγRIIIa/CD16 转染的 NK-92 细胞系,它们携带两种研究最多的 FCGR3A SNPs 的不同组合,代表了欧洲人群中的主要基因型。对这些细胞系的功能表征揭示了它们在ADCC和直接NK细胞毒性方面的差异,这可能会对设计使用NK细胞和肿瘤抗原导向的mAbs的癌症免疫疗法产生影响。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
NK Cytotoxicity Mediated by NK-92 Cell Lines Expressing Combinations of Two Allelic Variants for FCGR3.

Natural killer (NK) cells play an important role in the surveillance of viral infections and cancer. NK cell antibody-dependent cellular cytotoxicity (ADCC) and direct cytotoxicity are mediated by the recognition of antibody-coated target cells through the Fc gamma receptor IIIA (FcγRIIIa/CD16) and by ligands of activating/inhibitory NK receptors, respectively. Allelic variants of the FCGR3A gene include the high-affinity single-nucleotide polymorphism (SNP) rs396991 (V176F), which is associated with the efficacy of monoclonal antibody (mAb) therapies, and the SNP rs10127939 (L66H/R). The contribution of FCGR3A SNPs to NK cell effector functions remains controversial; therefore, we generated a panel of eight NK-92 cell lines expressing specific combinations of these SNPs and tested their cytotoxicities. NK-92 cells were stably transfected with plasmids containing different combinations of FCGR3A SNPs. Messenger RNA and FcγRIIIa/CD16 cell surface expressions were detected using new generation sequencing (NGS) and flow cytometry, respectively. All FcγRIIIa/CD16-transfected NK-92 cell lines exhibited robust ADCC against three different target cell lines with minor differences. In addition, enhanced direct NK cytotoxicity against K562 target cells was observed, suggesting a mechanistic role of FcγRIIIa/CD16 in direct NK cytotoxicity. In conclusion, we generated eight FcγRIIIa/CD16-transfected NK-92 cell lines carrying different combinations of two of the most studied FCGR3A SNPs, representing the major genotypes described in the European population. The functional characterization of these cell lines revealed differences in ADCC and direct NK cytotoxicity that may have implications for the design of adoptive cancer immunotherapies using NK cells and tumor antigen-directed mAbs.

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来源期刊
Antibodies
Antibodies IMMUNOLOGY-
CiteScore
7.10
自引率
6.40%
发文量
68
审稿时长
11 weeks
期刊介绍: Antibodies (ISSN 2073-4468), an international, peer-reviewed open access journal which provides an advanced forum for studies related to antibodies and antigens. It publishes reviews, research articles, communications and short notes. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. Full experimental and/or methodical details must be provided. Electronic files or software regarding the full details of the calculation and experimental procedure - if unable to be published in a normal way - can be deposited as supplementary material. This journal covers all topics related to antibodies and antigens, topics of interest include (but are not limited to): antibody-producing cells (including B cells), antibody structure and function, antibody-antigen interactions, Fc receptors, antibody manufacturing antibody engineering, antibody therapy, immunoassays, antibody diagnosis, tissue antigens, exogenous antigens, endogenous antigens, autoantigens, monoclonal antibodies, natural antibodies, humoral immune responses, immunoregulatory molecules.
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