Pediatric Hospital Bambino Gesù, Rome, Italy.
Department of Experimental Medicine and Center of Excellence for Biomedical Research, University of Genova, Genova, Italy.
J Allergy Clin Immunol. 2021 Jun;147(6):2343-2357. doi: 10.1016/j.jaci.2020.12.648. Epub 2021 Jan 23.
There is limited knowledge on the origin and development from CD34 precursors of the ample spectrum of human natural killer (NK) cells, particularly of specialized NK subsets.
This study sought to characterize the NK-cell progeny of CD34DNAM-1CXCR4 and of other precursors circulating in the peripheral blood of patients with chronic viral infections (eg, HIV, hepatitis C virus, cytomegalovirus reactivation).
Highly purified precursors were obtained by flow cytometric sorting and cultured in standard NK-cell differentiation media (ie, SCF, FLT3, IL-7, IL-15). Phenotypic and functional analyses on progenies were performed by multiparametric cytofluorimetric assays. Transcriptional signatures of NK-cell progenies were studied by microarray analysis. Inhibition of cytomegalovirus replication was studied by PCR.
Unlike conventional CD34 precursors, LinCD34DNAM-1CXCR4 precursors from patients with chronic infection, rapidly differentiate into cytotoxic, IFN-γ-secreting CD94/NKG2CKIRCD57 NK-cell progenies. An additional novel subset of common lymphocyte precursors was identified among LinCD34CD56CD16 cells and characterized by expression of CXCR4 and lack of perforin and CD94. LinCD34CD56CD16PerfCD94CXCR4 precursors are also endowed with generation potential toward memory-like NKG2CNK cells. Maturing NK-cell progenies mediated strong human cytomegalovirus-inhibiting activity. Microarray analysis confirmed a transcriptional signature compatible with NK-cell progenies and with maturing adaptive NK cells.
During viral infections, precursors of adaptive NK cells are released and circulate in the peripheral blood.
人们对 CD34DNAM-1CXCR4 前体细胞以及循环于慢性病毒感染（如 HIV、丙型肝炎病毒、巨细胞病毒再激活）患者外周血中的其他前体细胞向广泛谱系的人类自然杀伤（NK）细胞，尤其是向特定 NK 亚群分化的起源和发育过程知之甚少。
本研究旨在描述 CD34DNAM-1CXCR4 前体细胞和其他前体细胞在慢性病毒感染患者外周血中的 NK 细胞后代。
通过流式细胞术分选获得高纯度的前体细胞，并在标准 NK 细胞分化培养基（即 SCF、FLT3、IL-7、IL-15）中培养。通过多参数细胞荧光术分析对后代进行表型和功能分析。通过微阵列分析研究 NK 细胞后代的转录特征。通过 PCR 研究巨细胞病毒复制的抑制情况。
与传统的 CD34 前体细胞不同，慢性感染患者的 LinCD34DNAM-1CXCR4 前体细胞可迅速分化为具有细胞毒性、IFN-γ 分泌功能的 CD94/NKG2CKIRCD57 NK 细胞后代。在 LinCD34CD56CD16 细胞中还鉴定到一种新型的普通淋巴细胞前体细胞亚群，其特征为表达 CXCR4，缺乏穿孔素和 CD94。LinCD34CD56CD16PerfCD94CXCR4 前体细胞也具有向记忆样 NKG2CNK 细胞分化的潜能。成熟的 NK 细胞后代具有强烈的人巨细胞病毒抑制活性。微阵列分析证实了一种与 NK 细胞后代和成熟适应性 NK 细胞相兼容的转录特征。
在病毒感染期间，适应性 NK 细胞的前体细胞被释放并循环于外周血中。
