Yoon Hee-Soo, Kim Hee-Yeon, Cho Kyung-Ah, Kim Yu-Hee, Woo So-Youn, Kim Han-Su, Kang Jihee-Lee, Ryu Kyung-Ha, Park Joo-Won
Department of Biochemistry, College of Medicine, Ewha Womans University, Seoul 07084, Korea.
Department of Microbiology, College of Medicine, Ewha Womans University, Seoul 07084, Korea.
Biology (Basel). 2022 Feb 7;11(2):255. doi: 10.3390/biology11020255.
Reactive oxygen species (ROS) generated by neutrophils provide a frontline defence against invading pathogens. We investigated the supportive effect of tonsil-derived mesenchymal stem cells (TMSCs) on ROS generation from neutrophils using promyelocytic HL-60 cells. Methods: Differentiated HL-60 (dHL-60) cells were cocultured with TMSCs isolated from 25 independent donors, and ROS generation in dHL-60 cells was measured using luminescence. RNA sequencing and real-time PCR were performed to identify the candidate genes of TMSCs involved in augmenting the oxidative burst of dHL-60 cells. Transcriptome analysis of TMSCs derived from 25 independent donors revealed high levels of procollagen C-endopeptidase enhancer 2 (PCOLCE2) in TMSCs, which were highly effective in potentiating ROS generation in dHL-60 cells. In addition, knockdown in TMSCs abrogated TMSC-induced enhancement of ROS production in dHL-60 cells, indicating that TMSCs increased the oxidative burst in dHL-60 cells via PCOLCE2. Furthermore, the direct addition of recombinant PCOLCE2 protein increased ROS production in dHL-60 cells. These results suggest that PCOLCE2 secreted by TMSCs may be used as a therapeutic candidate to enhance host defences by increasing neutrophil oxidative bursts. PCOLCE2 levels in TMSCs could be used as a marker to select TMSCs exhibiting high efficacy for enhancing neutrophil oxidative bursts.
中性粒细胞产生的活性氧（ROS）为抵御入侵病原体提供了一线防御。我们使用早幼粒细胞HL-60细胞研究了扁桃体来源的间充质干细胞（TMSC）对中性粒细胞ROS生成的支持作用。方法：将分化的HL-60（dHL-60）细胞与从25个独立供体分离的TMSC共培养，并使用发光法测量dHL-60细胞中的ROS生成。进行RNA测序和实时PCR以鉴定参与增强dHL-60细胞氧化爆发的TMSC候选基因。对来自25个独立供体的TMSC进行转录组分析，发现TMSC中前胶原C端肽酶增强子2（PCOLCE2）水平较高，其在增强dHL-60细胞中的ROS生成方面非常有效。此外，TMSC中的敲低消除了TMSC诱导的dHL-60细胞中ROS产生的增强，表明TMSC通过PCOLCE2增加了dHL-60细胞中的氧化爆发。此外，直接添加重组PCOLCE2蛋白可增加dHL-60细胞中的ROS产生。这些结果表明，TMSC分泌的PCOLCE2可作为一种治疗候选物，通过增加中性粒细胞氧化爆发来增强宿主防御。TMSC中的PCOLCE2水平可作为选择对增强中性粒细胞氧化爆发具有高效能的TMSC的标志物。
