背景:越来越多的证据表明,脂肪组织来源的间充质干细胞(ADSCs)是管理2019年冠状病毒病(COVID-19)的有效治疗方法;然而,需要进一步阐明,以确定它们对T辅助细胞相关转录因子(TFs)mRNA表达和外周血单核细胞(PBMC)细胞因子释放的潜在免疫调节作用.
目的:探讨ADSCs对结直肠癌(CRC)合并重症COVID-19(CRC+)患者PBMC中TFsmRNA表达及细胞因子释放的影响。
方法:来自CRC+患者(PBMC-C+)和年龄匹配的CRC患者(PBMC-C)的PBMC在存在/不存在ADSC的情况下被刺激和培养。T-boxTFTBX21(T-bet)的mRNA水平,GATA结合蛋白3(GATA-3),RAR相关孤儿受体C(RORC),通过逆转录酶-聚合酶链反应测定PBMC中的叉头盒P3(FoxP3)。评估培养上清液的干扰素γ(IFN-γ)水平,白细胞介素4(IL-4),IL-17A,和使用酶联免疫吸附测定的转化生长因子β1(TGF-β1)。
结果:与PBMC-C相比,PBMC-C+表现出更高的T-bet和RORCmRNA水平,IFN-γ和IL-17A水平升高。此外,在PBMC-C+中观察到FoxP3mRNA和TGF-β1显著降低,T-bet/GATA-3、RORC/FoxP3、IFN-γ/IL-4和IL-17A/TGF-β1比值增加。此外,ADSCs显著诱导功能性调节性T细胞(Treg)亚群,FoxP3mRNA和TGF-β1释放水平的增加证明了这一点。这伴随着T-bet和RORC的mRNA水平显着降低,IFN-γ和IL-17A的释放,T-bet/GATA-3、RORC/FoxP3、IFN-γ/IL-4和IL-17A/TGF-β1比值,与单独的PBMC-C+相比。
结论:目前的体外研究表明,ADSCs对PBMC-C+有免疫抑制作用,有利于Treg反应。因此,基于ADSC的细胞疗法对于对常规疗法无反应的重度COVID-19患者可能是一种有益的方法。
BACKGROUND: Accumulating evidence has shown that adipose tissue-derived mesenchymal stem cells (ADSCs) are an effective therapeutic approach for managing coronavirus disease 2019 (COVID-19); however, further elucidation is required to determine their underlying immunomodulatory effect on the mRNA expression of T helper cell-related transcription factors (TFs) and cytokine release in peripheral blood mononuclear cells (PBMCs).
OBJECTIVE: To investigate the impact of ADSCs on the mRNA expression of TFs and cytokine release in PBMCs from colorectal cancer (CRC) patients with severe COVID-19 (CRC+ patients).
METHODS: PBMCs from CRC+ patients (PBMCs-C+) and age-matched CRC patients (PBMCs-C) were stimulated and cultured in the presence/absence of ADSCs. The mRNA levels of T-box TF TBX21 (T-bet), GATA binding protein 3 (GATA-3), RAR-related orphan receptor C (RORC), and forkhead box P3 (FoxP3) in the PBMCs were determined by reverse transcriptase-polymerase chain reaction. Culture supernatants were evaluated for levels of interferon gamma (IFN-γ), interleukin 4 (IL-4), IL-17A, and transforming growth factor beta 1 (TGF-β1) using an enzyme-linked immunosorbent assay.
RESULTS: Compared with PBMCs-C, PBMCs-C+ exhibited higher mRNA levels of T-bet and RORC, and increased levels of IFN-γ and IL-17A. Additionally, a significant decrease in FoxP3 mRNA and TGF-β1, as well as an increase in T-bet/GATA-3, RORC/FoxP3, IFN-γ/IL-4, and IL-17A/TGF-β1 ratios were observed in PBMCs-C+. Furthermore, ADSCs significantly induced a functional regulatory T cell (Treg) subset, as evidenced by an increase in FoxP3 mRNA and TGF-β1 release levels. This was accompanied by a significant decrease in the mRNA levels of T-bet and RORC, release of IFN-γ and IL-17A, and T-bet/GATA-3, RORC/FoxP3, IFN-γ/IL-4, and IL-17A/TGF-β1 ratios, compared with the PBMCs-C+alone.
CONCLUSIONS: The present in vitro studies showed that ADSCs contributed to the immunosuppressive effects on PBMCs-C+, favoring Treg responses. Thus, ADSC-based cell therapy could be a beneficial approach for patients with severe COVID-19 who fail to respond to conventional therapies.