Hardersen Randolf, Enebakk Terje, Christiansen Dorte, Bergseth Grethe, Brekke Ole-Lars, Mollnes Tom Eirik, Lappegård Knut Tore, Hovland Anders
Department of Nephrology, Division of Internal Medicine, Nordland Hospital, Bodø, Norway.
Research Laboratory, Nordland Hospital, Bodø, Norway.
APMIS. 2018 Apr;126(4):342-352. doi: 10.1111/apm.12821.
The aim of the study was to investigate the role of complement factor 5 (C5) in reactions elicited by plasma separation using blood from a C5-deficient (C5D) individual, comparing it to C5-deficient blood reconstituted with C5 (C5DR) and blood from healthy donors. Blood was circulated through an ex vivo plasma separation model. Leukocyte CD11b expression and leukocyte-platelet conjugates were measured by flow cytometry during a 30-min period. Other markers were assessed during a 240-min period. Granulocyte and monocyte CD11b expression did not increase in C5D blood during plasma separation. In C5DR samples granulocytes CD11b expression, measured by mean fluorescence intensity (MFI), increased from 10481 ± 6022 (SD) to 62703 ± 4936, and monocytes CD11b expression changed from 13837 ± 7047 to 40063 ± 713. Granulocyte-platelet conjugates showed a 2.5-fold increase in the C5DR sample compared to the C5D sample. Monocyte-platelet conjugates increased independently of C5. In the C5D samples, platelet count decreased from 210 × 10 /L (201-219) (median and range) to 51 × 10 /L (50-51), and C3bc increased from 14 CAU/mL (21-7) to 198 CAU/mL (127-269), whereas TCC formation was blocked during plasma separation. In conclusion, up-regulation of granulocyte and monocyte CD11b during plasma separation was C5-dependent. The results also indicate C5 dependency in granulocyte-platelet conjugates formation.
本研究的目的是调查补体因子5(C5)在使用C5缺陷(C5D)个体血液进行血浆分离引发的反应中的作用,并将其与用C5重构的C5缺陷血液(C5DR)以及健康供体的血液进行比较。血液通过体外血浆分离模型进行循环。在30分钟内通过流式细胞术测量白细胞CD11b表达和白细胞-血小板结合物。在240分钟内评估其他标志物。血浆分离过程中,C5D血液中的粒细胞和单核细胞CD11b表达未增加。在C5DR样本中,通过平均荧光强度(MFI)测量的粒细胞CD11b表达从10481±6022(标准差)增加到62703±4936,单核细胞CD11b表达从13837±7047变为40063±713。与C5D样本相比,C5DR样本中的粒细胞-血小板结合物增加了2.5倍。单核细胞-血小板结合物的增加与C5无关。在C5D样本中,血小板计数从210×10⁹/L(201 - 219)(中位数和范围)降至51×10⁹/L(50 - 51),C3bc从14 CAU/mL(21 - 7)增加到198 CAU/mL(127 - 269),而血浆分离过程中末端补体复合物(TCC)形成被阻断。总之,血浆分离过程中粒细胞和单核细胞CD11b的上调是C5依赖性的。结果还表明粒细胞-血小板结合物形成依赖于C5。