Myeloid-derived suppressor cell measurements in fresh and cryopreserved blood samples.
Myeloid-derived suppressor cell measurements in fresh and cryopreserved blood samples.
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DOI：
10.1016/j.jim.2012.04.004
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发表时间：
2012-07-31
影响因子：
2.2
通讯作者：
Whiteside, Theresa L.
中科院分区：
文献类型：
--
作者：
Kotsakis, Athanasios;Harasymczuk, Malgorzata;Schilling, Bastian;Georgoulias, Vasilis;Argiris, Athanassios;Whiteside, Theresa L.
髓源性抑制细胞（MDSC）存在于人外周血中，代表了具有单核细胞和粒细胞特征的异质细胞群。为了提供可靠评估MDSC频率和功能的指南，我们比较了从正常对照和癌症患者中获得的新鲜和冷冻外周血单个核细胞（PBMC）样本。从4名健康供体和21名癌症患者中获得PBMC。用标记抗体染色，用流式细胞术检测冷冻前后DR - /LIN - /CD11b+、DR - /LIN - /CD15+、DR - /LIN - /CD33+和DR - /low/CD14+细胞的频率。以cfse为基础的抑菌法检测MDSC的抑菌功能。流式细胞术检测MDSC亚群中精氨酸酶I的表达和活性氧（ROS）的上调。与DR - /LIN - /CD15+和DR - /LIN - /CD33+亚群相比，MDSC的DR - /low/CD14+和DR - /LIN - /CD11b+亚群对低温保存/解冻程序的抵抗力更强。后两个MDSC亚群的频率在冷冻保存后显著降低。除了DR−/LIN−/CD15+细胞外，所有细胞都抑制自体csf标记的CD4+细胞的增殖，但在冷冻保存后失去抑制活性。只有DR−/LIN−/CD15+细胞精氨酸酶I阳性，但冷冻保存后失去表达。体外刺激后，只有新鲜的DR - /LIN - /CD11b+和DR - /LIN - /CD15+细胞产生ROS。人类MDSC的研究应在新鲜血液样本中进行。如果样本必须冷冻保存，监测CD11b+和CD14+ MDSC亚群提供最可靠的结果。精氨酸酶I的表达或受刺激的ROS的产生通过流式细胞术评估是有用的标记，仅在新鲜样品的MDSC亚群。
Myeloid-derived suppressor cells (MDSC) present in the human peripheral blood, represent a heterogeneous population of cells with monocytic and granulocytic features. To provide guidelines for reliable assessments of the frequency and function of MDSC, we compared fresh vs. cryopreserved peripheral blood mononuclear cell (PBMC) samples obtained from normal controls and patients with cancer. PBMC were obtained from 4 healthy donors and 21 patients with cancer. They were stained with labeled antibodies, and the frequency of DR−/LIN−/CD11b+, DR−/LIN−/CD15+, DR−/LIN−/CD33+ and DR−/low/CD14+ cells was determined by flow cytometry before and after cryopreservation. CFSE-based suppressor assays were used to test inhibitory functions of MDSC. Arginase I expression and reactive oxygen species (ROS) upregulation in MDSC subsets were evaluated by flow cytometry. The DR−/low/CD14+ and DR−/LIN−/CD11b+ subsets of MDSC were found to be more resistant to the cryopreservation/thawing procedure compared to the DR−/LIN−/CD15+ and DR−/LIN−/CD33+ subsets. The frequency of the latter two MDSC subsets was significantly reduced after cryopreservation. All but DR−/LIN−/CD15+ cells inhibited proliferation of autologous CSFE-labeled CD4+ cells but lost suppressor activity after cryopreservation. Only DR−/LIN−/CD15+ cells were positive for Arginase I, but lost its expression after cryopreservation. Only fresh DR−/LIN−/CD11b+ and DR−/LIN−/CD15+ cells produced ROS after in vitro stimulation. Studies of human MDSC should be performed in fresh blood samples. If samples have to be cryopreserved, monitoring of CD11b+ and CD14+ MDSC subsets provides the most reliable results. Arginase I expression or stimulated ROS production assessed by flow cytometry are useful markers for MDSC subsets only in fresh samples.
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影响因子：
5.6
作者：
Greten TF;Manns MP;Korangy F
通讯作者：
Korangy F
影响因子：
2.8
作者：
Vuk-Pavlovic, Stanimir;Bulur, Peggy A.;Lin, Yi;Qin, Rui;Szumlanski, Carol L.;Zhao, Xinghua;Dietz, Allan B.
通讯作者：
Dietz, Allan B.
影响因子：
4.4
作者：
Nefedova, Y;Huang, M;Gabrilovich, D
通讯作者：
Gabrilovich, D
影响因子：
4.4
作者：
Almand, B;Clark, JI;Gabrilovich, DI
通讯作者：
Gabrilovich, DI
影响因子：
2.2
作者：
Elkord, Eyad
通讯作者：
Elkord, Eyad