Department of Laboratory Medicine, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, 540142 Targu Mures, Romania.
Department of Laboratory Medicine, Emergency Clinical County Hospital Targu Mures, 540136 Targu Mures, Romania.
Int J Mol Sci. 2022 Oct 6;23(19):11875. doi: 10.3390/ijms231911875.
The aim of the study was to evaluate the dynamic changes of the total Natural Killer (NK) cells and different NK subpopulations according to their differentiated expression of CD16/CD56 in COVID-19 patients. Blood samples with EDTA were analyzed on day 1 (admission moment), day 5, and day 10 for the NK subtypes. At least 30,000 singlets were collected for each sample and white blood cells were gated in CD45/SSC and CD16/CD56 dot plots of fresh human blood. From the lymphocyte singlets, the NK cells subpopulations were analyzed based on the differentiated expression of surface markers and classified as follows: CD16CD56/CD16CD56/CD16CD56/CD16CD56. By examining the CD56 versus CD16 flow cytometry dot plots, we found four distinct NK sub-populations. These NK subtypes correspond to different NK phenotypes from secretory to cytolytic ones. There was no difference between total NK percentage of different disease forms. However, the total numbers decreased significantly both in survivors and non-survivors. Additionally, for the CD16CD56 phenotype, we observed different patterns, gradually decreasing in survivors and gradually increasing in those with fatal outcomes. Despite no difference in the proportion of the CD16CD56 NK cells in survivors vs. non-survivors, the main cytokine producers gradually decline during the study period in the survival group, underling the importance of adequate IFN production during the early stage of SARS-CoV-2 infection. Persistency in the circulation of CD56 NK cells may have prognostic value in patients, with a fatal outcome. Total NK cells and the CD16CD56 NK subtypes exhibit significant decreasing trends across the moments for both survivors and non-survivors.
本研究旨在评估根据 COVID-19 患者 NK 细胞分化表达 CD16/CD56 的不同亚群，评估总自然杀伤 (NK) 细胞和不同 NK 亚群的动态变化。在第 1 天（入院时）、第 5 天和第 10 天，用 EDTA 处理的血液样本分析 NK 亚型。每个样本至少收集 30,000 个单细胞，并在新鲜人血的 CD45/SSC 和 CD16/CD56 点图中对白细胞进行门控。从淋巴细胞单细胞中，根据表面标志物的分化表达分析 NK 细胞亚群，并分类如下：CD16CD56+/CD16CD56+/CD16CD56+/CD16CD56+。通过检查 CD56 与 CD16 的流式细胞术点图，我们发现了四个不同的 NK 亚群。这些 NK 亚型对应于从分泌到细胞溶解的不同 NK 表型。不同疾病形式的总 NK 百分比没有差异。然而，幸存者和非幸存者的总数量均显著减少。此外，对于 CD16CD56 表型，我们观察到不同的模式，幸存者逐渐减少，而致命结局的患者逐渐增加。尽管幸存者与非幸存者的 CD16CD56 NK 细胞比例无差异，但在生存组中，研究期间主要细胞因子产生细胞逐渐减少，表明在 SARS-CoV-2 感染的早期阶段适当产生 IFN 的重要性。在患者中，CD56 NK 细胞的持续循环可能具有预后价值，具有致命结局。总 NK 细胞和 CD16CD56 NK 亚群在幸存者和非幸存者的所有时间点均表现出明显的下降趋势。
