体外应用HDAC抑制剂TSA诱导骨髓细胞分化为MDSC的实验研究  

The effect of HDAC enzyme inhibitor TSA on inducing bone marrow cells to differentiate into MDSC in vitro

作　　者：赵曙光[1] 高伟年[1] 于丁[1] 陈子英[1] ZHAO Shu-guang;GAO Wei-nian;YU Ding;CHEN Zi-ying(Department of Cardiac Surgery,the Second Affiliated Hospital of Hebei Medical University,Shijiazhuang050000,China)

机构地区：[1]河北医科大学第二医院心外科,石家庄050000

出　　处：《现代免疫学》2021年第2期94-98,共5页Current Immunology

基　　金：国家自然科学基金(81470068)。

摘　　要：为探讨利用广谱组蛋白去乙酰化酶(histone deacetylase, HDAC)抑制剂曲古抑菌素A(trichostatin A,TSA)调控骨髓(bone marrow, BM)细胞分化、发育及炎性因子转录,体外诱导BM细胞分化为髓源性抑制细胞(myeloid-derived suppressor cell, MDSC)的细胞群,应用TSA及重组小鼠GM-CSF(recombinant mouse GM-CSF,rmGM-CSF)体外培养BM细胞7 d后,用FACS检测MDSC相关抗原CD11b、Gr-1的表达,并通过ELISA检测培养上清液中炎性因子IL-6的分泌水平,经磁珠分选、纯化MDSC,行混合淋巴细胞培养,鉴定其免疫抑制活性。结果显示,TSA+rmGM-CSF可诱导BM细胞分化为纯度为59.7%的CD11b^(+)Gr-1^(+)MDSC细胞群,该细胞群经Gr-1^(+)磁珠分选可获得93.8%的CD11b^(+)Gr-1^(+)MDSC,混合淋巴细胞培养显示MDSC具有抑制异基因CD4^(+)CD25^(-)T细胞增殖的能力,该抑制活性弱于Treg。该研究提示,HDAC抑制剂TSA与GM-CSF联合应用可高效诱导BM细胞分化为MDSC。This in vitro study is aimed to induce bone marrow(BM)cells into myeloid-derived suppressor cells(MDSC)by histone deacetylase(HDAC)inhibitor trichostatin A(TSA)to regulate the differentiation and development of BM cells and the transcription of inflammatory factors.After the BM cells were cultured with TSA and recombinant mouse GM-CSF(rmGM-CSF)in vitro for 7 d,the expressions of MDSC-related CD11 b and Gr-1 were detected by FACS,and the secretion level of inflammatory factor IL-6 in the culture supernatant was detected by ELISA.MDSC were purified by Gr-1^(+)magnetic bead sorting and mixed lymphocyte culture was performed to identify their immunosuppressive activity.The results showed that TSA+rmGM-CSF could induce BM cells into a CD11b^(+)Gr-1^(+)MDSC cell population with a purity of 59.7%and the purity could augment to 93.8%by further Gr-1^(+)magnetic bead sorting.Mixed lymphocyte culture showed that MDSCs had the ability to inhibit the proliferation of allogeneic CD4^(+)CD25^(-)T cells,and the inhibitory activity was weaker than Treg.The results indicate that the combination of HDAC inhibitor TSA and GM-CSF can effectively induce BM cells to differentiate into MDSC.

关 键 词：组蛋白去乙酰化酶 骨髓细胞 髓源性抑制细胞 

分 类 号：R392.12[医药卫生—免疫学]