Induction of tissue factor expression in whole blood: Lack of evidence for the presence of tissue factor expression in granulocytes
Induction of tissue factor expression in whole blood: Lack of evidence for the presence of tissue factor expression in granulocytes
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DOI：
10.1055/s-0037-1613934
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发表时间：
2000-06-01
影响因子：
6.7
通讯作者：
Olsen, JO
中科院分区：
文献类型：
--
作者：
Osterud, B;Rao, LVM;Olsen, JO
采用Western印迹和ELISA法研究了血小板、肿瘤坏死因子(TNF)和佛波酯(PMA)对内毒素(LPS)诱导的组织因子(TF)活性和组织因子抗原的影响。脂多糖可诱导单核细胞表达相关水平的转铁蛋白抗原和活性。肿瘤坏死因子和PMA单独使用时，不能诱导单核细胞的转铁蛋白活性和抗原，但可使单核细胞内毒素诱导的转铁蛋白活性和抗原增加2~3倍。在分离的血细胞中加入富含血小板的血浆，可提高内毒素诱导的单核细胞的转铁蛋白活性，但不能提高单核细胞的抗原水平。与全血小板相比，血小板裂解物既能增强脂多糖诱导的转铁蛋白活性，又能增强抗原。从肝素化血浆中分离出的粒细胞单独或与PMA共同孵育2或24小时，均不能产生TF抗原或活性。虽然从与内毒素和PMA孵育24小时的全血中分离出的粒细胞制剂明显具有显著的TF活性和抗原，但这可能是由于微量污染单核细胞所致。在有粒细胞存在的情况下，血小板可上调脂多糖诱导的TF活性，但不能上调其抗原活性。提示这种增强的TF活性可能是由于单核细胞与活化的单核细胞融合后PS浓缩或血小板与活化的单核细胞的PS浓缩所致。
The present investigation was undertaken to explore the effect of platelets, tumor necrosis factor (TNF) and phorbel ester [phorbol 12-myristate 13-acetate (PMA)] on lipopolysaccharide (LPS)-induced tissue factor (TF) activity and TF antigen by using Western blot and ELISA-techniques. LPS was found to induce correlating levels of TF antigen and the activity in monocytes. TNF and PMA, when used alone, failed to induce TF activity and the antigen in monocytes, but enhanced the LPS-induced TF activity and the antigen by 2 to 3-fold. Addition of platelet rich plasma to isolated blood cells enhanced the LPS-induced TF activity but not the antigen levels in monocytes. In contrast to whole platelets, platelet lysates enhanced both LPS-in- duced TF activity and the antigen. Granulocytes isolated from heparinized plasma incubated for 2 or 24 h with LPS alone or together with PMA, failed to generate TF antigen or the activity. Although granulocyte preparations isolated from whole blood that was incubated for 24 h with LPS and PMA apparently possessed a significant amount of TF activity and the antigen, this could be accounted for by trace levels of contaminating monocytes. Upregulation of LPS-induced TF activity but not the antigen by platelets in the presence of granulocytes suggests that the increased TF activity could be the result of PS enrichment of monocytes by fusion or platelets with activated monocytes.