Van Aelst Britt, Devloo Rosalie, Vandekerckhove Philippe, Compernolle Veerle, Feys Hendrik B
Transfusion Research Center, Belgian Red Cross-Flanders, Ghent, Belgium.
Blood Service of the Belgian Red Cross-Flanders, Mechelen, Belgium.
Transfusion. 2015 Oct;55(10):2404-14. doi: 10.1111/trf.13137. Epub 2015 Apr 25.
Ultraviolet (UV) light illumination in the presence of exogenously added photosensitizers has been used to inactivate pathogens in platelet (PLT) concentrates for some time. The THERAFLEX UV-C system, however, illuminates PLT concentrates with UV-C light without additional photoactive compounds. In this study residual PLT function is measured in a comprehensive paired analysis of UV-C-treated, gamma-irradiated, and untreated control PLT concentrates.
A pool-and-split design was used with buffy coat-derived PLT concentrates in 65% SSP+ additive solution. Thrombus formation kinetics in microfluidic flow chambers onto immobilized collagen was investigated with real-time video microscopy. PLT aggregation, membrane markers, and cellular metabolism were determined concurrently.
Compared to gamma-treated and untreated controls, UV-C treatment significantly affected thrombus formation rates on Days 5 and 7, not Day 2. PLT degranulation (P-selectin) and PLT apoptosis (annexin V binding) was slightly but significantly increased from Day 2 on. UV-C treatment moreover induced integrin αIIb β3 conformational changes reminiscent of activation. However, subsequent integrin activation by either PAR1-activating hexapeptide (PAR1AP) or convulxin was unaffected. This was confirmed by PLT aggregation studies induced with collagen, PAR1AP, and ristocetin at two different agonist concentrations. Finally, UV-C slightly increased lactic acid production rates, resulting in significantly decreased pH on Days 5 and 7, but never dropped below 7.2.
UV-C pathogen inactivation treatment slightly but significantly increases PLT activation markers but does not profoundly influence activatability nor aggregation. The treatment does, however, attenuate thrombus formation kinetics in vitro in microfluidic flow chambers, especially after storage.
在外源性添加光敏剂的情况下，紫外线（UV）照射已被用于使血小板（PLT）浓缩物中的病原体失活一段时间。然而，THERAFLEX UV-C系统使用UV-C光照射PLT浓缩物，无需额外的光活性化合物。在本研究中，通过对UV-C处理、γ射线辐照和未处理的对照PLT浓缩物进行全面的配对分析，测量了残余的PLT功能。
采用池化和分割设计，使用来自 Buffy 层的PLT浓缩物，置于65% SSP+添加剂溶液中。通过实时视频显微镜研究微流体流动室中固定化胶原蛋白上的血栓形成动力学。同时测定PLT聚集、膜标志物和细胞代谢。
与γ处理和未处理的对照相比，UV-C处理在第5天和第7天显著影响血栓形成率，而不是第2天。从第2天开始，PLT脱颗粒（P-选择素）和PLT凋亡（膜联蛋白V结合）略有但显著增加。此外，UV-C处理诱导整合素αIIbβ3构象变化，类似于激活状态。然而，随后由PAR1激活六肽（PAR1AP）或芋螺毒素诱导的整合素激活不受影响。这通过在两种不同激动剂浓度下用胶原蛋白、PAR1AP和瑞斯托菌素诱导的PLT聚集研究得到证实。最后，UV-C略微提高了乳酸产生率，导致第5天和第7天pH值显著降低，但从未降至7.2以下。
UV-C病原体灭活处理略微但显著增加了PLT激活标志物，但并未深刻影响其激活能力或聚集。然而，该处理确实会减弱微流体流动室中体外血栓形成动力学，尤其是在储存后。
