Data_Sheet_1_Human Dental Pulp Stem Cells Modulate Cytokine Production in vitro by Peripheral Blood Mononuclear Cells From Coronavirus Disease 2019 Patients.doc

A subset of patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) developed a condition of hyper-inflammation, which can cause multi-organ damage and the more severe forms of coronavirus disease 2019 (COVID-19). Mesenchymal stem cells (MSCs) can promote tissue regeneration and modulate immune responses and, thus, have the rational requirements to be used to counteract SARS-CoV-2-induced pneumonia and hyper-inflammation. The aim of the present study was to gain insight into possible mechanisms of action of MSCs obtained from human dental pulp [dental pulp stem cells (DPSCs)] in COVID-19 patients. We investigated the concentrations of 18 cytokines in supernatants of peripheral blood mononuclear cells (PBMCs) obtained from COVID-19 patients cultured in vitro alone and in contact with DPSCs. The modulation of cytokines in PBMCs was confirmed by real-time PCR. IL-6 was the sole cytokine detected in supernatants of DPSCs. In resting conditions, co-culture increased IL-1β, IL-2, IL-5, IL-6, IL-10, IL-18, TNFα, and granulocyte macrophage colony-stimulating factor (GM-CSF) levels. When PBMCs were activated with anti-CD3/CD28 antibody-coated beads, co-culture increased IL-6 and GM-CSF, whereas it decreased IFNγ, TNFα, IL-2, IL-5, IL-9, IL-10, IL-12 (p70), IL-17A, IL-18, IL-21, IL-23, and IL-27 levels. Concentrations of IL-1β, IL-4, IL-13, and IL-22 were not affected. The comparison of cytokine concentrations in supernatants of PBMCs from COVID-19 patients vs. healthy subjects revealed lower concentrations of IL-10 and higher concentrations of IL-18 in supernatants of CD3/CD28-activated PBMCs from COVID-19 patients. Results are explorative but indicate that DPSCs can modulate the production of cytokines deregulated in COVID-19 patients, supporting their potential use in COVID-19.

部分感染严重急性呼吸综合征冠状病毒2（severe acute respiratory syndrome coronavirus 2，SARS-CoV-2）的患者会出现过度炎症状态，该状态可引发多器官损伤，并进展为更危重的2019冠状病毒病（coronavirus disease 2019，COVID-19）。间充质干细胞（mesenchymal stem cells，MSCs）可促进组织再生并调节免疫应答，因此具备用于对抗SARS-CoV-2诱导的肺炎与过度炎症的理论依据。本研究旨在阐明源自人牙髓的间充质干细胞[牙髓干细胞（dental pulp stem cells，DPSCs）]对COVID-19患者的潜在作用机制。本研究检测了COVID-19患者来源的外周血单个核细胞（peripheral blood mononuclear cells，PBMCs）单独体外培养、与DPSCs共培养两种条件下，其上清液中18种细胞因子的浓度水平。通过实时荧光定量PCR（real-time PCR）验证了PBMCs内细胞因子的调控变化。仅在DPSCs的上清液中检测到白细胞介素6（interleukin 6，IL-6）。在静息状态下，共培养可提升IL-1β、IL-2、IL-5、IL-6、IL-10、IL-18、肿瘤坏死因子α（tumor necrosis factor α，TNF-α）以及粒细胞-巨噬细胞集落刺激因子（granulocyte macrophage colony-stimulating factor，GM-CSF）的表达水平。当用抗CD3/CD28抗体包被磁珠激活PBMCs后，共培养可提升IL-6与GM-CSF的水平，同时降低干扰素γ（interferon γ，IFN-γ）、TNF-α、IL-2、IL-5、IL-9、IL-10、IL-12（p70）、IL-17A、IL-18、IL-21、IL-23以及IL-27的表达水平。IL-1β、IL-4、IL-13与IL-22的浓度未受共培养的影响。对比COVID-19患者与健康受试者PBMCs上清液的细胞因子浓度发现，经CD3/CD28激活的COVID-19患者PBMCs上清液中，IL-10浓度更低而IL-18浓度更高。本研究结果尚属探索性，但表明DPSCs可调控COVID-19患者体内失调的细胞因子生成，为其用于COVID-19的治疗提供了理论支持。