Shalekoff S, Page-Shipp L, Tiemessen C T
Medical Research Council AIDS Virus Research Unit, National Institute for Virology, Johannesburg, South Africa.
Clin Diagn Lab Immunol. 1998 Sep;5(5):695-702. doi: 10.1128/CDLI.5.5.695-702.1998.
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 degreesC for 18 h with or without the addition of the cytokines gamma interferon (IFN-gamma), granulocyte-macrophage colony-stimulating factor (GM-CSF), IFN-gamma plus GM-CSF, tumor necrosis factor beta, or interleukin-6. Of the cytokines tested, the combination of IFN-gamma 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.
采用全血模型评估温度和抗凝剂对外周血白细胞上活化标志物HLA - DR和CD11b表达的影响。用乙二胺四乙酸（EDTA）或肝素抗凝的静脉血取自6名健康献血者和13名住院患者（8名1型人类免疫缺陷病毒血清阳性并发肺结核患者和5名肺炎患者）。对两名正常献血者的全血进行了初步评估，细胞立即用于HLA - DR和CD11b标志物染色，或在室温或37℃孵育18小时后染色，孵育时添加或不添加细胞因子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或白细胞介素 - 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）上标志物表达的调节作用最为显著，尤其是HLA - DR表达，其检测需要诱导。因此，在考虑疾病存在情况下上述影响的进一步评估中使用了这些细胞因子。结果表明，全血中PMN和淋巴细胞上活化标志物的表达受孵育温度、抗凝剂选择的影响，观察到的影响取决于：（i）特定的细胞表面标志物；（ii）所研究的细胞类型；（iii）疾病的存在与否。因此，建议对用于评估表型或免疫功能的体外全血模型针对上述影响进行仔细评估。
