粒细胞-巨噬细胞集落刺激因子调节失代偿期肝硬化合并脓毒症患者髓源性抑制细胞和 Treg 活性。

Granulocyte-Macrophage Colony-Stimulating Factor Modulates Myeloid-Derived Suppressor Cells and Treg Activity in Decompensated Cirrhotic Patients With Sepsis.

机构信息

Department of Molecular and Cellular Medicine, Institute of Liver and Biliary Sciences, New Delhi, India.

Amity Institute of Biotechnology, Amity University, Noida, India.

出版信息

Front Immunol. 2022 Jun 3;13:828949. doi: 10.3389/fimmu.2022.828949. eCollection 2022.

BACKGROUND

Decompensated cirrhosis patients are more prone to bacterial infections. Myeloid-derived suppressor cells (MDSCs) expand in sepsis patients and disrupt immune cell functions. Granulocyte-macrophage colony-stimulating factor (GM-CSF) therapy helps in restoring immune cell functions and resolving infections. Its role in MDSC modulation in cirrhosis with sepsis is not well understood.

METHODS

A total of 164 decompensated cirrhotic-62 without (w/o), 72 with sepsis, and 30 with sepsis treated with GM-CSF-and 15 healthy were studied. High-dimensional flow cytometry was performed to analyze MDSCs, monocytes, neutrophils, CD4 T cells, and Tregs at admission and on days 3 and day 7. co-cultured MDSCs with T cells were assessed for proliferation and apoptosis of T cells and differentiation to Tregs. Plasma factors and mRNA levels were analyzed by cytokine-bead assay and qRT-PCR.

RESULTS

Frequencies of MDSCs and Tregs were significantly increased ( = 0.011 and = 0.02) with decreased CD4 T cells ( = 0.01) in sepsis than w/o sepsis and healthy controls (HCs) ( = 0.000, = 0.07, and = 0.01) at day 0 and day 7. In sepsis patients, MDSCs had increased IL-10, Arg1, and iNOS mRNA levels ( = 0.016, = 0.043, and = 0.045). co-cultured MDSCs with T cells drove T-cell apoptosis ( = 0.03, = 0.03) with decreased T-cell proliferation and enhanced FOXP3 expression ( = 0.044 and = 0.043) in sepsis compared to w/o sepsis at day 0. Moreover, blocking the MDSCs with inhibitors suppressed FOXP3 expression. GM-CSF treatment in sepsis patients significantly decreased MDSCs and FOXP3 Tregs but increased CD4 T-cell functionality and improved survival.

CONCLUSION

MDSCs have an immunosuppressive function by expanding FOXP3 Tregs and inhibiting CD4 T-cell proliferation in sepsis. GM-CSF treatment suppressed MDSCs, improved T-cell functionality, and reduced Tregs in circulation.

背景

代偿失调性肝硬化患者更易发生细菌感染。髓系来源的抑制细胞（MDSCs）在脓毒症患者中扩增，并破坏免疫细胞功能。粒细胞-巨噬细胞集落刺激因子（GM-CSF）治疗有助于恢复免疫细胞功能和解决感染。但其在肝硬化合并脓毒症中对 MDSC 调节的作用尚不清楚。

方法

共纳入 164 例失代偿性肝硬化患者-62 例无（w/o）脓毒症，72 例脓毒症和 30 例脓毒症接受 GM-CSF 治疗-以及 15 例健康对照者。入院时和第 3 天及第 7 天进行高维流式细胞术分析 MDSCs、单核细胞、中性粒细胞、CD4 T 细胞和 Tregs。共培养 MDSCs 与 T 细胞，评估 T 细胞增殖和凋亡以及分化为 Tregs 的情况。通过细胞因子珠试验和 qRT-PCR 分析血浆因子和 mRNA 水平。

结果

与无脓毒症和健康对照组相比，脓毒症患者（=0.011 和=0.02）MDSCs 和 Tregs 频率显著升高（=0.01），CD4 T 细胞（=0.01）减少（=0.000，=0.07，和=0.01）在第 0 天和第 7 天。脓毒症患者的 MDSCs 中 IL-10、Arg1 和 iNOS mRNA 水平升高（=0.016，=0.043，和=0.045）。与无脓毒症患者相比，在第 0 天，共培养 MDSCs 与 T 细胞后，T 细胞凋亡增加（=0.03，=0.03），T 细胞增殖减少，FOXP3 表达增强（=0.044 和=0.043）。此外，用抑制剂阻断 MDSCs 可抑制 FOXP3 表达。GM-CSF 治疗脓毒症患者可显著减少 MDSCs 和 FOXP3 Tregs，但可增强 CD4 T 细胞功能，并改善存活率。