a Center of Perioperative Thrombosis and Hemostasis , Zurich , Switzerland.
b Institute of Laboratory Diagnostics, Hygiene and Transfusion Medicine , Academic City Hospital , Ludwigshafen , Germany.
Platelets. 2019;30(6):720-727. doi: 10.1080/09537104.2018.1513471. Epub 2018 Sep 11.
Platelet quality in different platelet concentrates (PCs) has been the subject of several studies. Nonetheless, there is a lack of robust data on the correlation and agreement among platelet function tests as a prerequisite for the association of PC functionality in vitro with platelet function in vivo post PC transfusion. The purpose of our study was to correlate a larger panel of platelet function assays in PCs and to assess whether the methods agree sufficiently and can be used interchangeably. Twelve apheresis platelet concentrates in plasma (APC), 16 pooled platelet concentrates in plasma (PPC), and 12 PPC in T-sol (PPCA) were examined on days 1 and 4 after production. PCs were tested for platelet count, light transmission aggregation (LTA) induced by ADP, collagen, or TRAP; platelet ATP release induced by collagen; and spontaneous and ADP and TRAP-induced increase in CD62P and PAC1 expression measured by flow cytometry. All tests were performed in undiluted platelet-rich plasma, recalcified and mixed with an inhibitor of factor Xa and thrombin. Most platelet function parameters correlated significantly with each other, but agreement among methods was insufficient. A proper inverse correlation was observed between ADP-induced LTA and spontaneous platelet activation assessed by CD62P expression ( = -0.61, < 0.0001). Spontaneous CD62P correlated also significantly with spontaneous PAC1 ( = 0.69, < 0.0001) and inversely with TRAP-induced CD62P expression ( = -0.86, < 0.0001). We found significant correlations among all flow cytometric assays measuring platelet CD62P and PAC1 expression induced by ADP or TRAP. Subsequent Bland Altman analysis revealed insufficient agreement between methods. With one exception (collagen-induced LTA compared with TRAP-induced LTA, percentage error = 16%) the limits of agreement expressed as percentage error exceeded the chosen acceptable difference of 30%. In APC, platelet count was 41% and 44% higher, respectively, than in PPC and PPCA ( < 0.0001). Spontaneous CD62P and PAC1 expression were significantly greater, and ADP-induced aggregation and agonist-induced increase in CD62P and PAC1 were significantly lower in PPCA compared to APC and PPC on day 4 of storage. ADP and TRAP-induced CD62P and PAC1 activatability fell significantly during storage between day 1 and day 4 in APC and PPCA, but not in PPC. In conclusion, different platelet function tests capture different aspects of platelet function and do not correlate and agree sufficiently to be used interchangeably.
不同血小板浓缩物(PC)中的血小板质量一直是多项研究的主题。尽管如此,对于血小板功能测试之间的相关性和一致性,缺乏强有力的数据,这是将 PC 体外功能与 PC 输注后体内血小板功能相关联的前提。我们的研究目的是在 PC 中对更大的血小板功能检测面板进行相关性分析,并评估这些方法是否足够一致且可以互换使用。在制备后第 1 天和第 4 天,对 12 份富含血小板的血浆中的血小板浓缩物(APC)、16 份富含血小板的血浆中的血小板浓缩物(PPC)和 12 份 T-sol 中的血小板浓缩物(PPCA)进行了检测。PC 被检测血小板计数、ADP、胶原或 TRAP 诱导的透光率聚集(LTA);胶原诱导的血小板 ATP 释放;以及通过流式细胞术测量的自发和 ADP 及 TRAP 诱导的 CD62P 和 PAC1 表达增加。所有测试均在未稀释的富含血小板的血浆中进行,重钙化并与因子 Xa 和凝血酶抑制剂混合。大多数血小板功能参数彼此之间具有显著相关性,但方法之间的一致性不足。ADP 诱导的 LTA 与通过 CD62P 表达评估的自发性血小板激活之间存在适当的负相关性(r=-0.61,p<0.0001)。自发性 CD62P 也与自发性 PAC1 显著相关(r=0.69,p<0.0001),与 TRAP 诱导的 CD62P 表达呈负相关(r=-0.86,p<0.0001)。我们发现所有流式细胞术检测方法之间均存在显著相关性,这些方法可测量 ADP 或 TRAP 诱导的血小板 CD62P 和 PAC1 表达。随后的 Bland Altman 分析表明,这些方法之间的一致性不足。除了一种情况(与 TRAP 诱导的 LTA 相比,胶原诱导的 LTA,百分比误差=16%)之外,以百分比误差表示的一致性界限超过了所选的 30%可接受差异。在 APC 中,血小板计数分别比 PPC 和 PPCA 高 41%和 44%(p<0.0001)。在储存第 4 天,与 APC 和 PPC 相比,PPCA 中的自发性 CD62P 和 PAC1 表达明显更高,ADP 诱导的聚集和激动剂诱导的 CD62P 和 PAC1 表达增加明显更低。在 APC 和 PPCA 中,ADP 和 TRAP 诱导的 CD62P 和 PAC1 可激活性在第 1 天至第 4 天的储存期间显著下降,但在 PPC 中没有下降。总之,不同的血小板功能检测方法捕捉到了血小板功能的不同方面,并且彼此之间没有足够的相关性和一致性,无法互换使用。