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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Effects of Anticoagulants and Temperature on Expression of Activation Markers CD11b and HLA-DR on Human Leukocytes
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Effects of Anticoagulants and Temperature on Expression of Activation Markers CD11b and HLA-DR on Human Leukocytes

机译：抗凝剂和温度对人白细胞激活标记CD11b和HLA-DR表达的影响

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A whole-blood model was used to evaluate the effects of temperature and anticoagulant on the expression of activation markers HLA-DR and CD11b on peripheral leukocytes. Venous blood, anticoagulated with either EDTA or heparin, was obtained from six healthy blood donors and 13 hospitalized patients (8 human immunodeficiency virus type 1-seropositive individuals with concurrent pulmonary tuberculosis and 5 patients with pneumonia). A preliminary evaluation was carried out with whole blood from two of the normal donors, and cells were stained immediately for HLA-DR and CD11b markers or stained after incubation at room temperature or 37°C for 18 h with or without the addition of the cytokines gamma interferon (IFN-γ), granulocyte-macrophage colony-stimulating factor (GM-CSF), IFN-γ plus GM-CSF, tumor necrosis factor beta, or interleukin-6. Of the cytokines tested, the combination of IFN-γ and GM-CSF had the most pronounced modulation of marker expression on polymorphonuclear neutrophils (PMN), in particular, HLA-DR expression, which required induction for its detection. These cytokines were therefore used in further evaluations that considered the above-mentioned effects in the presence of disease. Results indicated that the expression of activation markers on PMN and lymphocytes in whole blood are influenced by the temperature of incubation and the choice of anticoagulant and the effects noted were dependent on (i) the particular cell surface marker, (ii) the cell type being studied, and (iii) the presence or absence of disease. It is therefore recommended that ex vivo whole-blood models for evaluating phenotype or immune function be carefully evaluated for the above-mentioned effects.
机译：使用全血模型评估温度和抗凝剂对外周血白细胞激活标记HLA-DR和CD11b表达的影响。从6名健康献血者和13例住院患者（8例人类免疫缺陷病毒1型血清阳性并发肺结核的患者和5例肺炎患者）获得了用EDTA或肝素抗凝的静脉血。用来自两个正常供体的全血进行初步评估，并立即对细胞进行HLA-DR和CD11b标记染色，或者在室温或37°C下孵育18小时（添加或不添加细胞因子）后进行染色γ干扰素（IFN-γ），粒细胞巨噬细胞集落刺激因子（GM-CSF），IFN-γ加GM-CSF，肿瘤坏死因子β或白介素6。在所测试的细胞因子中，IFN-γ和GM-CSF的组合对多形核中性粒细胞（PMN）的标志物表达，尤其是HLA-DR表达具有最明显的调节作用，需要对其进行诱导才能检测到。因此，将这些细胞因子用于考虑到疾病存在时上述效应的进一步评估中。结果表明，温育温度和抗凝剂的选择会影响全血中PMN和淋巴细胞上活化标记的表达，所观察到的效果取决于（i）特定的细胞表面标记，（ii）细胞类型（iii）是否存在疾病。因此，建议仔细评估用于评估表型或免疫功能的离体全血模型的上述作用。

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