Decidua-derived granulocyte macrophage colony-stimulating factor induces polymorphonuclear myeloid-derived suppressor cells from circulating CD15+ neutrophils
Decidua-derived granulocyte macrophage colony-stimulating factor induces polymorphonuclear myeloid-derived suppressor cells from circulating CD15+ neutrophils
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蜕膜源性粒细胞巨噬细胞集落刺激因子诱导循环 CD15 中性粒细胞产生多形核骨髓源性抑制细胞
DOI：
10.1093/humrep/deaa217
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发表时间：
2020-12-01
影响因子：
6.1
通讯作者：
Zhao, Aimin
中科院分区：
文献类型：
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作者：
Li, Congcong;Chen, Chao;Zhao, Aimin
研究问题：早孕期蜕膜衍生因子是否刺激循环中性粒细胞向多形核粒细胞骨髓衍生抑制细胞（PMN-MDSC）转化？总结回答：循环中性粒细胞可以获得PMN-MDSC样表型和功能通过磷酸化的信号转导和转录激活因子5/程序性死亡配体2（pSTAT 5/PD-L2）信号转导后与蜕膜衍生的粒细胞巨噬细胞集落刺激因子（GM-CSF）的刺激。中性粒细胞具有高度的异质性和可塑性，在刺激后可转化为免疫抑制性PMN-MDSC。为了分析髓源性抑制细胞（MDSC）亚群比例，从非妊娠妇女收集12个子宫内膜组织和12个外周血样品，40例蜕膜组织和16例外周血标本均来自正常早孕妇女，这些妇女因非医学原因接受了随孕龄变化的选择性手术终止妊娠6-10周。收集29个蜕膜组织用于分离CD 15（+）PMN-MDSC。收集20个子宫内膜组织和30个蜕膜组织用于细胞因子分析、免疫组织化学或中性粒细胞刺激。对象/材料、地点、方法：采用流式细胞术检测正常早孕妇女蜕膜、外周血、子宫内膜及非妊娠妇女外周血中MDSC的比例。采用流式细胞术比较蜕膜PMN-MDSC和循环中性粒细胞的表型和功能。用蜕膜外植体上清液（DES）刺激循环中性粒细胞，用流式细胞术和免疫荧光法测定表型。蜕膜PMN-MDSC和DES条件中性粒细胞的抑制能力通过流式细胞术分析，有或没有抗程序性细胞死亡-1（PD-1）抗体。通过Luminex测定法检测DES和子宫内膜外植体上清液（EES）中的细胞因子。用ELISA法和免疫组化法检测GM-CSF的表达。用DES、EES、DES加抗GM-CSF抗体或EES加GM-CSF刺激中性粒细胞。CD 11b，凝集素样氧化低密度脂蛋白受体-1（LOX-1），PD-L2和pSTAT 5的表达通过流式细胞仪测定。主要结果和机会的作用：与非妊娠妇女外周血，非妊娠妇女的子宫内膜或外周血相比，早孕蜕膜中的PMN-MDSCs的频率显著增加。蜕膜PMN-MDSC抑制T细胞增殖和细胞因子产生。蜕膜PMN-MDSC的表型与成熟活化的中性粒细胞相似。DES诱导中性粒细胞中的CD 11b、LOX-1、PD-L2表达和STAT 5磷酸化。中性粒细胞中的PD-L2表达依赖于STAT 5磷酸化。蜕膜PMN-MDSC和DES调节的中性粒细胞均通过PD-1信号传导抑制T细胞增殖。GM-CSF在蜕膜中表达上调，并诱导中性粒细胞表达CD 11b、LOX-1和PD-L2。DES显著诱导CD 11b、LOX-1、PD-L2表达和STAT 5磷酸化。抗GM-CSF抗体可显著阻断中性粒细胞的这种刺激作用。EES未诱导CD 11b、LOX-1、PD-L2表达或STAT 5磷酸化，而GM-CSF处理充分刺激中性粒细胞中的CD 11b、LOX-1、PD-L2表达和STAT 5磷酸化。这项研究是基于体外实验，我们不能评估中性粒细胞分化为PMN-MDSC在其他网站之前，进入母体，由于人体样本的可用性有限，结果的更广泛意义：这是第一项研究证明了蜕膜PMN-MDSC是一组具有成熟状态的免疫调节细胞，并且中性粒细胞可以通过pSTAT 5/PD-L2信号转导被蜕膜来源的GM-CSF诱导为PMN-MDSC样表型。本研究表明GM-CSF可通过调节PMN-MDSCs促进早孕免疫耐受，进而为GM-CSF在妊娠并发症的防治提供了潜在的作用。
STUDY QUESTION: Do decidua-derived factors stimulate the conversion of circulating neutrophils to polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) in early human pregnancy?SUMMARY ANSWER: Circulating neutrophils can acquire PMN-MDSC-like phenotypes and function via phosphorylated signal transducer and activator of transcription 5/programmed death ligand 2 (pSTAT5/PD-L2) signalling after stimulation with decidua-derived granulocyte macrophage colony-stimulating factor (GM-CSF).WHAT IS KNOWN ALREADY: PMN-MDSCs are an important immunoregulatory cell type in early pregnancy. Neutrophils are of high heterogeneity and plasticity and can polarize to immunosuppressive PMN-MDSCs upon stimulation.STUDY DESIGN, SIZE, DURATION: For analysis of myeloid-derived suppressor cell (MDSC) subset proportions, 12 endometrium tissues and 12 peripheral blood samples were collected from non-pregnant women, and 40 decidua tissues and 16 peripheral blood samples were obtained from women with normal early pregnancy undergoing elective surgical pregnancy termination for nonmedical reasons with gestation age of 6-10 weeks. Twenty-nine decidua tissues were collected for isolation of CD15(+) PMN-MDSCs. Twenty endometrium tissues and 30 decidua tissues were collected for cytokine analysis, immunohistochemistry or neutrophil stimulation. Peripheral blood samples were obtained from 36 healthy donors for isolation of CD3(+) T cells and CD15(+) neutrophils.PARTICIPANTS/MATERIALS, SETTING, METHODS: The proportion of MDSC subsets in the decidua and peripheral blood of normal early pregnancy, endometrium and peripheral blood of non-pregnant women was analysed by flow cytometry. The phenotypes and function of decidual PMN-MDSCs and circulating neutrophils were compared by flow cytometry. Circulating neutrophils were stimulated with decidual explant supernatant (DES) and the phenotypes were measured by flow cytometry and immunofluorescence. The suppressive capacity of decidual PMN-MDSCs and DES-conditioned neutrophils was analysed by flow cytometry with or without antiprogrammed cell death-1 (PD-1) antibody. Cytokines from DES and endometrial explant supernatant (EES) were detected by a Luminex assay. GM-CSF expression was determined by ELISA and immunohistochemistry. Neutrophils were stimulated with DES, EES, DES with anti-GM-CSF antibody or EES with GM-CSF. CD11b, lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), PD-L2 and pSTAT5 expression were measured by flow cytometry.MAIN RESULTS AND THE ROLE OF CHANCE: The frequency of PMN-MDSCs was significantly increased in the decidua of early pregnancy compared with peripheral blood of non-pregnant women, the endometrium of non-pregnant women or peripheral blood during early pregnancy. Decidual PMN-MDSCs suppressed T-cell proliferation and cytokine production. Phenotypes of decidual PMN-MDSCs were similar to mature activated neutrophils. DES-induced CD11b, LOX-1, PD-L2 expression and STAT5 phosphorylation in neutrophils. The PD-L2 expression in neutrophils was dependent on STAT5 phosphorylation. Both decidual PMN-MDSCs and DES-conditioned neutrophils suppressed T-cell proliferation via PD-1 signalling. GM-CSF was up-regulated in the decidua and induced CD11b, LOX-1 and PD-L2 expression on neutrophils. DES significantly induced CD11b, LOX-1, PD-L2 expression and STAT5 phosphorylation. Anti-GM-CSF antibody remarkably blocked such stimulation in neutrophils. EES did not induce CD11b, LOX-1, PD-L2 expression or STAT5 phosphorylation, while GM-CSF treatment sufficiently stimulated CD11b, LOX-1, PD-L2 expression and STAT5 phosphorylation in neutrophils.LARGE SCALE DATA: N/ALIMITATIONS, REASONS FOR CAUTION: The study was based on in vitro experiments and we were not able to evaluate neutrophils differentiation to PMN-MDSCs in other sites before entering the maternal-foetal interface due to the limited availability of human samples. This needs to be explored using murine models.WIDER IMPLICATIONS OF THE FINDINGS: This is the first study demonstrating that decidual PMN-MDSCs are a group of immunoregulatory cells with mature status, and that neutrophils can be induced to a PMN-MDSC-like phenotype with decidua-derived GM-CSF via pSTAT5/PD-L2 signalling. This study indicates that GM-CSF can facilitate immune tolerance of early pregnancy through regulating PMN-MDSCs and further provides a potential role of GM-CSF in prevention and treatment for pregnancy complications.