Montanucci Pia, Pescara Teresa, Greco Alessia, Francisci Daniela, Basta Giuseppe, Calafiore Riccardo
Division of Internal Medicine and Endocrine and Metabolic Sciences (MISEM), Laboratory for Endocrine Cell Transplants and Biohybrid Organs, Department of Medicine and Surgery, University of Perugia Piazzale Gambuli I-06132, Perugia, Italy.
Division of Infectious Diseases, Department of Medicine and Surgery, University of Perugia Piazzale Gambuli I-06132, Perugia, Italy.
Am J Stem Cells. 2021 Aug 15;10(3):36-52. eCollection 2021.
The recent newly appeared Coronavirus disease (COVID-19), caused by an enveloped RNA virus named "severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)", is associated with severe respiratory morbidity and mortality. Recent studies have shown that lymphopenia and a cytokine mass release represent important pathogenic features, with clinical evidence of dyspnea and hypoxemia, often leading to acute respiratory distress syndrome (ARDS), in severely ill patients, with a high death toll. Currently, stem cells are actively being investigated for their potential use in many "untreatable" diseases. In this regard and in particular, Mesenchymal Stem Cells (MSC), due to their intrinsic features, including either ability to impact on regulation of the immune system, or association with both anti-viral and anti-inflammatory properties, or potential for differentiation into several cell lineages, have become a promising tool for cell and molecular-based therapies. On this background, we wished to explore whether human umbilical cord-derived mesenchymal stem cells (hUCMS) would represent a potential viable therapeutic approach for the management of critically ill COVID19 patients.
We tested the hUCMS effects on peripheral blood mononuclear cell (PBMCs) retrieved from patients with COVID19 (Ethical Committee CEAS Umbria, Italy CER N°3658/20 7, May, 2020), both as free cell monolayers and after envelopment in sodium alginate microcapsules. Both cell systems, after priming with IFN-γ, proved able to produce several immunomodulatory molecules such as IDO1 and HLAG5, although only the microencapsulated hUCMS were associated with massive and dose-dependent production of these factors.
The microencapsulated hUCMS improved allo-suppression in mixed lymphocytes reactions (MLRs), while also blunting T helper 1 and T helper 17 responses, that are involved with the cytokine storm and greatly contribute to the patient death. Moreover, we observed that both free and microencapsulated hUCMS permitted 5 days survival of in vitro culture maintained PBMCs extracted from very ill patients.
We have provided evidence that microencapsulated hUCMS in vitro, seem to represent a powerful tool to impact on several immune pathways, clearly deranged in COVID19 patients. Further study is necessary to begin in vivo assessment of this experimental system, upon determining both, the most appropriate time of the disease onset for intervention, and cell dosage/patient of our experimental product.
最近新出现的冠状病毒病（COVID-19）由一种名为“严重急性呼吸综合征冠状病毒2（SARS-CoV-2）”的包膜RNA病毒引起，与严重的呼吸系统发病率和死亡率相关。最近的研究表明，淋巴细胞减少和细胞因子大量释放是重要的致病特征，临床证据显示重症患者出现呼吸困难和低氧血症，常导致急性呼吸窘迫综合征（ARDS），死亡率很高。目前，干细胞正因其在许多“无法治疗”疾病中的潜在用途而受到积极研究。在这方面，特别是间充质干细胞（MSC），由于其内在特性，包括影响免疫系统调节的能力、与抗病毒和抗炎特性的关联以及分化为多种细胞谱系的潜力，已成为基于细胞和分子疗法的一种有前景的工具。在此背景下，我们希望探讨人脐带间充质干细胞（hUCMS）是否可能成为治疗重症COVID-19患者的一种可行治疗方法。
我们测试了hUCMS对从COVID-19患者（意大利翁布里亚地区伦理委员会CEAS，CER编号3658/20，2020年5月）获取的外周血单个核细胞（PBMC）的影响，细胞以游离单层形式以及包裹在海藻酸钠微胶囊中两种形式进行测试。两种细胞系统在经IFN-γ预处理后，均能产生多种免疫调节分子，如IDO1和HLAG5，不过只有微胶囊化的hUCMS与这些因子的大量且剂量依赖性产生相关。
微胶囊化的hUCMS改善了混合淋巴细胞反应（MLR）中的同种异体抑制，同时也减弱了辅助性T细胞1和辅助性T细胞17反应，这两种反应与细胞因子风暴有关，对患者死亡有很大影响。此外，我们观察到游离和微胶囊化的hUCMS都能使从重症患者提取的PBMC在体外培养中存活5天。
我们提供的证据表明，体外微胶囊化的hUCMS似乎是影响COVID-19患者中明显紊乱的多种免疫途径的有力工具。在确定疾病发作的最合适干预时间以及我们实验产品的细胞剂量/患者剂量后，有必要进一步开展该实验系统的体内评估研究。
