Richter Jillian R, Sutton Jeffrey M, Hexley Phillip, Johannigman Taylor A, Lentsch Alex B, Pritts Timothy A
Department of Surgery and Institute for Military Medicine, University of Cincinnati, Cincinnati, Ohio.
Shriners Hospitals for Children, Cincinnati, Ohio.
J Surg Res. 2018 Mar;223:128-135. doi: 10.1016/j.jss.2017.09.052. Epub 2017 Dec 22.
Leukoreduction prior to packed red blood cell (pRBC) storage is not a universally accepted practice. Our laboratory has previously shown that microvesicles (MVs) accumulate in pRBC units during storage and play an important role in lung injury after resuscitation. Currently, the effect of leukoreduction on MV formation in stored pRBC units is unknown. In the present study, we investigated the hypothesis that leukoreduction of pRBC units prior to storage would attenuate the production of MVs and decrease pulmonary inflammation after hemorrhage and resuscitation.
Leukoreduced and nonleukoreduced pRBC units were prepared from human donors and C57/Bl6 mice and stored for up to 42 d and 14 d, respectively. At intervals during storage, MVs were isolated from pRBC units, quantified and characterized based on size, morphology, and levels of proinflammatory cytokines. In additional experiments, mice underwent controlled hemorrhage followed by resuscitation with normal saline (NS) with or without equal numbers of MVs isolated from leukoreduced or nonleukoreduced stored mouse pRBC. Histologic lung sections were evaluated for the presence of tissue edema and inflammatory cells.
For both human and mouse pRBCs, the number of MVs significantly increased throughout the storage period. There were significantly fewer MVs present in leukoreduced units. The average MV size significantly increased over time and was similar between groups. Levels of interleukin 1α (IL-1α), regulated on activation, normal T cell expressed and secreted (RANTES), and macrophage-derived chemokine (MDC) were lower in MVs from leukoreduced pRBC units as compared with MVs from nonleukoreduced units. Hemorrhaged mice resuscitated with NS with the addition of MV from leukoreduced pRBC demonstrated significantly less pulmonary edema and inflammatory cell recruitment as compared to those resuscitated with NS with the addition of MV from nonleukoreduced pRBC.
Prestorage leukoreduction of pRBC units reduces the formation and proinflammatory properties of MV, which in turn decreases lung injury secondary to MV from stored pRBC units after hemorrhage and resuscitation.
在储存浓缩红细胞（pRBC）之前进行白细胞滤除并非普遍接受的做法。我们实验室先前已表明，微泡（MVs）在pRBC储存过程中会积聚，并在复苏后的肺损伤中起重要作用。目前，白细胞滤除对储存的pRBC单位中MV形成的影响尚不清楚。在本研究中，我们调查了以下假设：在储存前对pRBC单位进行白细胞滤除会减弱MV的产生，并减少出血和复苏后的肺部炎症。
从人类供体和C57/Bl6小鼠制备白细胞滤除和未进行白细胞滤除的pRBC单位，分别储存长达42天和14天。在储存期间的不同时间点，从pRBC单位中分离出MVs，根据大小、形态和促炎细胞因子水平进行定量和表征。在额外的实验中，小鼠经历控制性出血，然后用生理盐水（NS）进行复苏，添加或不添加从白细胞滤除或未进行白细胞滤除的储存小鼠pRBC中分离出的等量MVs。对肺组织切片进行评估，以确定是否存在组织水肿和炎症细胞。
对于人类和小鼠pRBC，在整个储存期间MV的数量显著增加。白细胞滤除的单位中存在的MV明显较少。平均MV大小随时间显著增加，且各组之间相似。与未进行白细胞滤除的单位的MV相比，白细胞滤除的pRBC单位的MV中白细胞介素1α（IL-1α）、活化调节正常T细胞表达和分泌因子（RANTES）以及巨噬细胞衍生趋化因子（MDC）的水平较低。与添加未进行白细胞滤除的pRBC的MV的NS复苏的出血小鼠相比，添加白细胞滤除的pRBC的MV的NS复苏的出血小鼠的肺水肿和炎症细胞募集明显减少。
储存前对pRBC单位进行白细胞滤除可减少MV的形成和促炎特性，进而减少出血和复苏后储存的pRBC单位的MV继发的肺损伤。
