Belge Kai-Uwe, Dayyani Farshid, Horelt Alexia, Siedlar Maciej, Frankenberger Marion, Frankenberger Bernhard, Espevik Terje, Ziegler-Heitbrock Löms
Institute for Immunology, University of Muenchen, Germany.
J Immunol. 2002 Apr 1;168(7):3536-42. doi: 10.4049/jimmunol.168.7.3536.
In human blood two monocyte populations can be distinguished, i.e., the CD14(++)CD16(-)DR(+) classical monocytes and the CD14(+)CD16(+)DR(++) proinflammatory monocytes that account for only 10% of all monocytes. We have studied TNF production in these two types of cells using three-color immunofluorescence and flow cytometry on whole peripheral blood samples stimulated with either LPS or with the bacterial lipopeptide S-(2,3-bis(palmitoyloxy)-(2-RS)-propyl)-N-palmitoyl-(R)-Cys-(S)-Ser-(S)-Lys(4)-OH,trihydrochloride (Pam3Cys). After stimulation with LPS the median fluorescence intensity for TNF protein was 3-fold higher in the proinflammatory monocytes when compared with the classical monocytes. After stimulation with Pam3Cys they almost exclusively responded showing 10-fold-higher levels of median fluorescence intensity for TNF protein. The median fluorescence intensity for Toll-like receptor 2 cell surface protein was found 2-fold higher on CD14(+)CD16(+)DR(++) monocytes, which may explain, in part, the higher Pam3Cys-induced TNF production by these cells. When analyzing secretion of TNF protein into the supernatant in PBMCs after depletion of CD16(+) monocytes we found a reduction of LPS-induced TNF by 28% but Pam3Cys-induced TNF was reduced by 64%. This indicates that the minor population of CD14(+)CD16(+) monocytes are major producers of TNF in human blood.
在人类血液中，可以区分出两种单核细胞群体，即CD14(++)CD16(-)DR(+)经典单核细胞和CD14(+)CD16(+)DR(++)促炎单核细胞，后者仅占所有单核细胞的10%。我们使用三色免疫荧光和流式细胞术，对用脂多糖（LPS）或细菌脂肽S-(2,3-双（棕榈酰氧基）-（2-RS）-丙基）-N-棕榈酰-（R）-半胱氨酸-（S）-丝氨酸-（S）-赖氨酸（4）-OH，三盐酸盐（Pam3Cys）刺激的全外周血样本中的这两种细胞类型的肿瘤坏死因子（TNF）产生情况进行了研究。用LPS刺激后，促炎单核细胞中TNF蛋白的中位荧光强度比经典单核细胞高3倍。用Pam3Cys刺激后，它们几乎完全有反应，TNF蛋白的中位荧光强度水平高10倍。在CD14(+)CD16(+)DR(++)单核细胞上发现Toll样受体2细胞表面蛋白的中位荧光强度高2倍，这可能部分解释了这些细胞中Pam3Cys诱导的TNF产生较高的原因。在去除CD16(+)单核细胞后分析外周血单核细胞（PBMC）中超滤液中TNF蛋白的分泌情况时，我们发现LPS诱导的TNF减少了28%，但Pam3Cys诱导的TNF减少了64%。这表明CD14(+)CD16(+)单核细胞的少数群体是人类血液中TNF的主要产生者。
