添加剂红细胞悬液保存期间炎性细胞因子水平动态观察

目的　观察添加剂红细胞悬液 (PRC)保存期间白细胞介素 (IL) 1 β、IL 6、IL 8和肿瘤坏死因子 (TNF) α水平变化 ,探讨非溶血性发热反应 (FNHTR)可能发生的机制 ;观察滤除白细胞对上述细胞因子水平的影响 ,为选择滤除白细胞时间提供依据。方法　 1 0U添加剂红细胞各分为 2等份 ,其中 1份滤除白细胞 ,分别于 0周和 1 ,3 ,5周末采用ELISA法测定IL 1 β、IL 6、IL 8和TNF α水平。 结果　新鲜PRC中IL 1 β、IL 6、IL 8和TNF α含量较低 ,随保存时间延长增高 ,尤以 3周后明显 ;保存前滤除白细胞后PRC保存期间IL 1 β、IL 6、IL 8和TNFα水平无明显升高 ;保存期末 4种炎性细胞因子水平与保存前WBC数量间呈正相关。结论　未去除白细胞的PRC在保存期间能够自发产生和积聚IL 1 β、IL 6、IL 8和TNF α ,部分FNHTR的发生可能与此有关 ;保存前滤除白细胞可防止这种现象发生     

滤除白细胞对红细胞悬液保存期间炎性细胞因子水平的影响及与临床非溶血性发热的关系

目的 :研究红细胞悬液保存期间白细胞介素IL 1,IL 6,IL 8和肿瘤坏死因子 (TNF α)水平变化和过滤对细胞因子水平及对非溶血性发热 (FNHTR)发生的影响。方法 :取 2U红细胞悬液 ,1U红细胞悬液予滤除白细胞和 1U红细胞悬液不滤除白细胞处理 ,保存 5周。分别在 0周、1周、3周、5周测定各细胞因子水平 ,临床观察过滤组和非过滤组FNHTR发生率。结果 :未经白细胞滤除的红细胞悬液随保存期延长各细胞因子水平逐渐升高 ,0周、1周、3周有统计学差异 (P <0 0 5 ) ,但 3周与 5周未见明显差异 (P >0 0 5 ) ,而经白细胞滤除的红细胞悬液保存期内各细胞因子的增加无统计学差异 (P >0 0 5 )。未滤除白细胞红细胞悬液输注后FNHTR发生率高于滤除组(P <0 0 5 )。结论 :未滤除白细胞的红细胞悬液在保存期内各细胞因子会积聚增多 ,可能与FNHTR发生有关 ,滤除后可有效防止细胞因子的增加 ,有助于降低临床FNHTR发生率     

白细胞滤器滤除血液中白细胞的时效性及临床意义

目的了解血液在保存期不同时间内炎性细胞因子(IL-1、IL-6、IL-8和TNF-α)含量的差异,探讨炎性细胞因子含量与非溶血性发热反应(FNHTR)的发生率的关系,为选择滤除白细胞时机提供依据。方法取合格红细胞悬液800袋(每袋2U),每袋无菌分装2等份(即每袋1U),其中一份滤除白细胞为过滤组,另一份未滤除白细胞为未滤组。分别于≤7天、8~14天、15~21天、≥22天不同时间段,测定血细胞指标和炎性细胞因子(IL-1、IL-6、IL-8和TNF-α)水平,并观察两组血液在不同保存期内临床输血发热反应(FNHTR)。结果过滤组在保存期内炎性细胞因子(IL-1、IL-6、IL-8和TNF-α)水平无明显升高。未滤组炎性细胞因子(IL-1、IL-6、IL-8和TNF-α)含量随保存时间延长而增高,尤以15~21天时间后更为明显。两组血液在不同保存期内临床输血发热反应(FNHTR)有明显差异。结论未滤组血液在保存期间能自发产生和积聚炎性细胞因子(IL-1、IL-6、IL-8和TNF-α),非溶血性发热反应(FNHTR)除同种白细胞抗体所引起外,可能与炎性细胞因子(IL-1、IL-6、IL-8和TNF-α)有关。血液保存最好当天或15天内滤除白细胞,可有效地减少临床输血FNHTR发生。     

应用滤除白细胞输血器输注血小板的临床护理分析

目的 为测试PL型滤器对浓缩血小板制剂(PCs)中白细胞的滤除作用。方法 在临床进行40例次伴血小板减少的血液病患者输注滤除白细胞的PCs。测定滤前、后PCs的血小板数、白细胞数。结果 血小板回收率为(91.5±7.8)％,白细胞滤除率为99.9％,输注无效率分别为16.25％(Donna标准)和8.75％(Eernise标准)。临床出血征象消失和好转率为93％。结论 PL型白细胞滤器具有高血小板回收、高白细胞去除的特点,是PCs滤除白细胞的理想滤器。     

保存前滤除白细胞对预防FNHTR及改善红细胞变形能力的作用

目的探讨推广血站型滤器的应用效果。方法将悬浮红细胞分为过滤组和对照组,在保存前和保存期检测5种细胞因子的水平及红细胞变形指数（EI）;临床输血患者按所输去白细胞悬浮红细胞过滤前保存时间分为5组,分别为0d、≤7d、≤14d、≤21d和〉21d,观察各组FNHTR发生率。结果对照组细胞因子随保存时间的延长而在血液中不断积累,过滤组细胞因子在保存中维持在原有的较低水平;在保存后期过滤组EI明显高于对照组;FNHTR发生率与过滤前保存时间呈正相关（P〈0．01）。结论在悬浮红细胞保存中存在多种细胞因子的产生和累积,而去白细胞过滤可在一定程度上抑制这一效应;保存前滤除白细胞可以有效预防FNHTR的发生,故悬浮红细胞应在保存前滤除白细胞。     

白细胞过滤对冰冻保存浓缩血小板制剂细胞因子和血小板功能的影响

目的探讨白细胞过滤对浓缩血小板悬液(platelet concentrate suspend,PCs)在冰冻保存前后的一些细胞因子及血小板体外功能的影响。方法取25份浓缩血小板样品(40ml/份),将每份样品再等分为4份,其中2份白细胞过滤(过滤组),另2份不过滤(未过滤组);将过滤组和未过滤组的分别常规保存和冰冻保存。常规保存0、1、3d和冰冻保存3个月解冻后0、1、3d的所有样品均采用ELISA法测定IL-2、IL-6、INFγ-、TNF-α、IL-10的含量;对冰冻保存复溶后当天的样品和新鲜样品分别作血小板聚集功能、粘附功能及血小板第Ⅲ因子活性检测;采用配对t检验作统计分析。结果未过滤组PCs冰冻保存后复溶0d与常规保存0d的PCs细胞因子的水平无明显升高,两种条件保存后1、3d细胞因子的水平均显著升高;过滤组PCs在冰冻保存和常规保存时细胞因子均无显著变化。过滤组和未过滤组PCs经冰冻保存后的血小板体外功能活性均无明显变化。结论PCs中的细胞因子在常规保存及冰冻保存复溶后随时间延长呈显著性增加趋势,白细胞过滤可明显减轻这种效应。白细胞过滤对冰冻保存的血小板体外功能活性无明显影响。     

去白细胞单采血小板的临床应用

【目的】研究保存前白细胞过滤与保存后白细胞过滤对单采血小板临床输注效果及非溶血性发热输血反应（FNHTR）发生率的影响。【方法】46例肿瘤化疗后血小板减少患者，分别输注保存前过滤单采血小板（保存前组）与保存后过滤单采血小板（保存后组），输注后1h及24h后检测外周血小板计数，以校正血小板计数增殖（CCI）判定输注效果，并考查FNHTR发生率。【结果】保存前组与保存后组其1h及24hCCI值无明显差异（P〉0．05），但保存前组FNHTR发生率要明显低于保存后组（P〈0．05）。【结论】两种方法制备的单采血小板均能有效地治疗血小板减少，但保存前过滤能更有效地减少FNHTR的发生率。     

一种制备乏白细胞的浓缩血小板的方法

长期输注血小板可致输血小板失效,已证明这是由于血小板中加杂白细胞引起HLA抗体所致,本文报道两种除去浓缩血小板(PCs)中白细胞的方法.用常规方法制备血小板,滤前将6份PCs混合.经醋酸纤维素(CA)滤器过滤的血小板,滤前须用前列腺素E_2(PGl_2)作短暂失活处理;经棉毛     

过滤和γ照射对保存期间单采血小板浓缩物中的RANTES和β1转化生长因子累积的影响

背景 在血小板保存过程中,由于血小板的活化,血小板衍生的生物学反应修饰因子(BRMs),包括RANTES和β1转化生长因子(TGF),在血小板成分中累积。它们可能与非溶,血性发热反应有关。在日本,大多数高品质的PCs来自单个供者的单采。研究设计与方法作者对采自单个供者的单采血小板浓缩物(单采PCs)单位和全血(白膜PCs)制备的血小板单位的RANTES和TGF-β1水平进行了研究。检测了保存前和保存后,过滤和γ照射对单采PCs保存期间上清液中RANTES和TGF-β1水平的影响。结果在保存的0至5天,RANTES和TGF-β1水平升高。RANTES和TGF-β1水平与血小板浓度相关(P<0.01),但与单采PCs中未经过滤减少白细胞的残余白细胞的浓度无关(P>0.05)。此外,RANTES和TGF-β1水平间有相关性(P<0.01)。使用阴电荷滤器过滤的单采PCs和经γ照射的单采PCs,在保存期内的任何时间,两种BRMs水平均与那些未经过滤     

亚低温对多发伤患者血清中细胞因子的影响及临床意义

目的探讨亚低温对多发性创伤患者血清中肿瘤坏死因子 α(TNF α)、白细胞介素 1β(IL 1β)、白细胞介素 6 (IL 6 )、白细胞介素 10 (IL 10 )的影响及临床意义。方法采用酶联免疫吸附法 (ELISA)监测 38例多发性创伤患者在亚低温治疗期间外周血清TNF α、IL 1β、IL 6、IL 10在伤后即刻、d3 、d5、d9的动态变化 ,并以 30例常规治疗的多发性创伤患者作为对照组进行对比分析。结果亚低温治疗组患者血清TNF α、IL 1β、IL 6、IL 10浓度比常规治疗组明显降低 (P <0 .0 5 )。结论亚低温治疗能够降低多发性创伤患者血清中细胞因子浓度 ,从而推测可能会减少患者多脏器功能不全综合征发生率     

Effect of filtration of platelet concentrates on the accumulation of cytokines and platelet release factors during storage

BACKGROUND: Platelet transfusions are frequently accompanied by febrile nonhemolytic transfusion reactions. These may be due, in part, to the release of cytokines interleukin 1 beta (IL-1 beta), interleukin 6 (IL- 6), interleukin 8 (IL-8), and tumor necrosis factor alpha (TNF-alpha) by white cells (WBCs) into the plasma during storage of platelet concentrates (PCs). Acting as endogenous pyrogens, these agents may induce inflammatory responses. STUDY DESIGN AND METHODS: This study proposed to determine if WBC reduction in PCs by filtration significantly reduced the levels of cytokines normally generated during storage of unfiltered PCs up to 5 days. Serotonin, platelet-derived growth factor (PDGF-AB), and von Willebrand factor levels were also assessed to establish whether or not filtration or storage elicited significant platelet activation and granule release. RESULTS: Filtration significantly reduced total WBC counts by 99.1 percent before storage (p < 0.001) without affecting total platelet counts. Compared to unfiltered PCs, filtration prevented a rise in the levels of each cytokine by Day 3 for IL-1 beta (27.7 vs. 0.6 pg/mL; p < 0.05), IL-6 (114.2 vs. 0.4 pg/mL; p < 0.001), and IL-8 (4.2 vs. 0.02 ng/mL; p < 0.001). By Day 5, further increases in the levels of all cytokines were noted in unfiltered PCs, but Day 0 levels remained in filtered PCs (IL-1 beta: 105.4 vs. 0.4 pg/mL, p < 0.001; TNF-alpha: 42.2 vs. 7.5 pg/mL, p < 0.025; IL-6: 268.8 vs. 0.4 pg/mL, p < 0.001; and IL-8: 7.6 vs. 0.02 ng/mL, p < 0.001). From Day 0 to Day 5, there were significant increases in serotonin (21.3 vs. 6.3 ng/mL, p < 0.05), PDGF-AB (72.6 vs. 25.8 ng/mL, p < 0.001), and von Willebrand factor (4.7 vs. 2.7 IU/mL, p < 0.05) in unfiltered PCs, with similar increased levels being observed in filtered PCs during storage. CONCLUSION: These data indicate that the accumulation of high levels of cytokines in stored PCs could be prevented by WBC-reduction filtration of PCs without the induction of significant platelet activation or granule release. As cytokines have the potential to induce febrile nonhemolytic transfusion reactions in patients, the transfusion of WBC-reduced PCs would be expected to reduce the frequency and severity of such reactions.     

Cytokines in WBC-reduced apheresis PCs during storage: a comparison of two WBC-reduction methods

BACKGROUND: Several studies have suggested that cytokine accumulation during storage of platelet concentrates (PCs) may mediate nonhemolytic febrile transfusion reactions and that a reduction in WBC numbers prevents the generation of cytokines. Despite efforts to minimize WBC contamination in apheresis PCs, high numbers of WBCs and increased cytokine levels may still occur, depending on the quality of the apheresis device employed. STUDY DESIGN AND METHODS: This study was undertaken to investigate whether PCs collected with WBC-reduction devices (Spectra LRS, COBE;or MCS+ LDP, Haemonetics) were sufficiently depleted of WBCs to limit cytokine accumulation during storage. The study evaluated 1) the levels of cytokines of WBC and platelet origin in two types of apheresis PCs during storage and 2) the effects of prestorage filtration on cytokine levels in the Spectra LRS PCs. RESULTS: In the Spectra LRS PCs, low levels of IL-6, IL-8, and monotype chemoattractant protein 1 (MCP-1) were detected in Day 1 PCs, and they remained consistent during the shelf life. RANTES, platelet factor 4 (PF4), beta-thromboglobulin (beta-TG), and transforming growth factor (TGF)-beta1 were also detected in these PCs, and their levels increased significantly on storage. Prestorage filtration of Spectra LRS PCs did not further reduce the levels of IL-6, IL-8, MCP-1, PF4, beta-TG, and TGF-beta1 in the filtered component. In the MCS+ LDP PCs, IL-6 was detected on Day 1, and its level increased significantly on storage, whereas the levels in the Spectra PCs remained steady. IL-8 levels were lower in MCS+ LDP PCs than in Spectra LRS PCs of the same age. MCP-1 levels were similar in both products on Day 1 and marginally increased in stored MCS+ LDP PCs. Substantial amounts of RANTES, PF4, beta-TG, and TGF-beta1 occurred in Day 1 MCS+ LDP PCs, and, on storage, these levels rose significantly. CONCLUSION: Despite a significant reduction in levels of WBC-derived cytokines, platelet-derived cytokines were present in different amounts in the two products.     

Cytokine accumulation in photochemically treated and gamma-irradiated platelet concentrates during storage

BACKGROUND: Photochemical treatment (PCT) for pathogen reduction of platelet concentrates (PCs) affects all cells containing DNA and/or RNA. Soluble mediators, which may cause transfusion reactions, are determined by the balance between secretion and/or cell destruction and binding and/or degradation. STUDY DESIGN AND METHODS: Ten double-dose single-donor leukoreduced PCs were split in two identical units. Two study arms were created: Study Arm A consisting of five PCT PCs with corresponding untreated control PCs and Study Arm B consisting of five PCT PCs with corresponding gamma-irradiated control PCs. PCs that had added PAS-III (Intersol) were treated with amotosalen and ultraviolet A light. Corresponding controls PCs, to which PAS-II (T-sol) were added, received no treatment or were gamma-irradiated before storage. Platelet (PLT)-derived (CCL5/RANTES, CXCL4/PF4, CCL3/MIP-1alpha, transforming growth factor [TGF]-beta, CXCL8/interleukin [IL]-8, IL-1beta) as well as white blood cell (WBC)-associated (IL-6, IL-10, IL-11, IL-12, tumor necrosis factor, interferon-gamma) cytokines were investigated by enzyme-linked immunosorbent assay and cytometric bead array during storage for up to 12 days. RESULTS: Independent of previous treatment we observed that all concentrates showed low levels of WBC-associated cytokines. PLT-derived cytokines were detected at higher levels and showed significant increase during storage. Statistical analysis showed lower PLT content per unit in PCT PCs, higher levels of activation variables in PCT PCs, and higher levels and accumulation rate of CCL5, CXCL4, TGF-beta, and CXCL8 in PCT PCs. CONCLUSION: PLTs are the main source of released cytokines during storage of untreated, gamma-irradiated, and PCT PCs. PCT may affect the level of PLT-derived cytokines in PCs. No additional reduction of WBC-associated cytokines were observed after PCT in prestorage leukoreduced PCs.     

The effect of prestorage WBC reduction on the rates of febrile nonhemolytic transfusion reactions to platelet concentrates and RBC

BACKGROUND: Febrile non-hemolytic transfusion reactions (FNHTRs) are a common complication of platelet concentrate (PC) and RBC transfusions, usually ascribed to cytokines released by WBCs and perhaps the platelets themselves during storage. Prestorage WBC reduction should abrogate the accumulation of these cytokines reducing the number of FNHTRs. STUDY DESIGN AND METHODS: A retrospective analysis of FNHTR to PCs and RBCs before universal WBC reduction (PrUR) (July 1997-January 1998 for PCs, July 1997-July 1999 for RBCs) and after its introduction (PoUR) (February 1998-August 2001 for PC, August 1999-August 2001 for RBCs) was undertaken. All transfusion reactions were stratified based on component and date of reaction. Other adverse transfusion reactions were grouped into three periods: July 1997-January 1998, February 1998-July 1999, and August 1999-August 2001. A chi-square test was performed to determine the significance of the differences between groups. RESULTS: In the PRUR group, there were: 231 FNHTRs in 70,396 RBC units transfused (0.33%) and 29 FNHTRs in 6502 PC units transfused (0.45% percent). In the PoUR group, there were 136 FNHTRs in 72,949 RBC units transfused (0.19%, p < 0.001) and 56 FNHTRs in 50,555 PC units transfused (0.11%, p < 0.001). Of the other adverse events, only TRALI reactions were significantly reduced. CONCLUSION: Prestorage WBC reduction significantly reduced the rate of FNHTRs to PCs and RBCs.     

Cytokines and adhesion molecules in the course of acute myocardial infarction

The plasma levels of interleukin 1 beta (IL 1β), interleukin 6 (IL 6), interleukin 8 (IL 8), tumor necrosis factor alpha (TNF-), E-selectin, ICAM 1 and C-reactive protein (CRP) have been studied in 24 patients with acute myocardial infarction in the course of 96 h. The plasma IL 1β and IL 6 levels were continually elevated during the 96 h study period (the peak of plasma IL 1β level was 22.2 pg/ml, S.D. 8.6, P<0.001, normal values of IL 1β are less than 10 pg/ml, the mean peak plasma concentration of IL 6 was 184.9 pg/ml, S.D. 134.7, vs. normal values of 15.57 pg/ml, S.D. 2.4, P<0.001). The mean plasma IL 8 level was increased for the duration of the study, the mean plasma IL 8 level was 103.0 pg/ml, S.D. 23.4 (normal value was below 30 pg/l, S.D. 8.0) P<0.001. The plasma TNF- level was elevated throughout the time of observation without any significant peak. The mean plasma TNF- concentration was 46.8 pg/ml, S.D. 2.13, vs. normal value 4.35 pg/ml, S.D. 1.23, P<0.001. The plasma E-selectin level reached the mean level of 145.1 ng/ml, S.D. 75.4, vs. normal value 29.1–63.4 ng/ml, P<0.001 at an interval of 15–42 h after the onset of the symptoms. The plasma ICAM 1 level showed only a slight significant increase during the first 36 h. The plasma CRP concentration increased later than IL 6, and reached a peak at 42 h after the onset of the symptoms (69.2 mg/l, S.D. 29.9, vs. 1.2 mg/l, S.D. 4.7, P<0.0001). We conclude that cytokines and adhesion molecules can play an important role in the mechanisms of tissue injury in the process of ischemia and reperfusion.     

White cell subsets in apheresis and filtered platelet concentrates

BACKGROUND: White cell (WBC)-reduced platelet concentrates (PCs) are defined by their absolute WBC count, a criterion which provides no information regarding the various WBC subsets contained in the PC. These heterogeneous cells are known to mediate different physiologic and pathophysiologic functions and account for distinct adverse transfusion responses. This study describes a method which allows the detection and quantification of these subsets and characterizes their presence in a variety of platelet components. STUDY DESIGN AND METHODS: Random-donor pooled PCs (RD PCs) and single-donor apheresis PCs (SD PCs) were studied. RD PCs consisting of 6 units of 2- to 3-day old PCs were randomly assigned to be filtered with one of four WBC-reduction filters from three different manufacturers (n=34). The residual WBCs were pelleted by centrifugation and isolated on a density gradient. The various WBC subsets were quantified by flow cytometry in unfiltered and filtered PCs using fluorescence and two-angle light scatter. SD PCs obtained with two manufacturer's systems and three processing protocols (n=30) were studied in like manner. RESULTS: WBC counts for non-WBC- reduced PCs averaged 3 × 10(8) in RD PCs and ranged from 8.6 to 9.6 × 10(6) per SD PC. Residual WBC counts in filtered PCs ranged from 2.3 × 10(4) to 2.2 × 10(5) and those in WBC-reduced SD PCs averaged 2.2 × 10(5) per unit. The data demonstrate significant phenotypic differences among PCs produced with various procedures. All SD PCs and two of four filtered RD PCs contained five WBC populations including granulocytes and monocytes, while RD PCs filtered with the remaining manufacturer's devices contained only lymphocytes. CONCLUSION: The data confirm that distinct phenotypic differences exist among PCs prepared with different devices and/or procedures. It is suggested that as for non-generic pharmaceuticals, the clinical benefits of these various PCs should be individually proved.     

WBC content of platelet concentrates prepared by the buffy coat method using different processing procedures and storage solutions

BACKGROUND: The number of WBCs in platelet concentrates (PCs) prepared by the buffy coat (BC) method with different storage solutions can result in low (5 x 10(6)/unit) WBC levels by the use of careful centrifugation techniques without filtration. At present, most blood banks use filtration steps to meet these requirements. The difference in processing methods and suspension solutions prompted the investigation of the influence of the various procedures on the WBC and platelet content of PCs. STUDY DESIGN AND METHODS: PCs from 5 BCs were harvested without or with inline filtration (AutoStop BC, Pall Corp.) in either plasma (PCs-plasma) or platelet additive solution (PCs-PAS-2). After preparation, samples were taken for counting WBCs and platelets and for analyzing WBC subsets by flow cytometry using specific MoAbs. The WBCs were concentrated before analysis of the WBC subsets. Results less than 2.5 cells per microL were considered below the limit of accuracy of the subset analysis. RESULTS: All filtered PCs met the AABB standard of 5 x 10(6) per unit and the European guidelines of 1 x 10(6) per unit. None of the nonfiltered PCs met the European guidelines, but all met the AABB guidelines. All filtered units gave residual WBC counts below the detection limit for subset analysis. Filtered PCs-plasma gave significantly higher platelet counts than filtered PCs-PAS-2 or nonfiltered PCs (p<0.01, ANOVA). CONCLUSION: Careful centrifugation of pooled BCs, with plasma or PAS-2, can result in PCs with low WBC contamination levels. However, filtered PCs are superior, because of better WBC removal and higher platelet counts.