库存血小板血浆中sCD40L含量变化及其对PMNs呼吸爆发的作用  

The effect of soluble CD40 ligand accumulated during platelet storage on polymorphonuclear neutrophils respiratory burst

作　　者：李玮[1,2] 谢如锋[2] 任亚娜[2] 杨懿铭[2] 钱开诚[2] 范华骅[2] 

机构地区：[1]华东师范大学生命科学学院生物医学系,上海200062 [2]上海市血液中心血液工程研究室

出　　处：《中国输血杂志》2013年第5期430-434,共5页Chinese Journal of Blood Transfusion

基　　金：国家自然科学基金(81270650);上海市公共卫生重点学科建设项目"输血医学"(12GWZX0201);上海市卫生局科研基金(20114294)

摘　　要：目的探讨单采血小板(A-PLTs)和白膜法浓缩血小板(BC-PLTs)库存过程中,其乏血小板血浆(简称血浆)中可溶性CD40配体(sCD40L)含量变化及对中性粒细胞(PMNs)呼吸爆发的作用。方法取库存d1、d3、d5的A-PLTs和BC-PLTs,离心分离获得血浆;ELISA检测血浆中sCD40L的含量;用活性氧特异性荧光探针二氢若丹明123(DHR),流式细胞术检测PMNs呼吸爆发;以小鼠抗人CD154抗体抑制试验去除sCD40L。结果 sCD40L含量(pg/mL),在血小板库存d1、d3、d5时,A-PLTs血浆分别为1 341.62±279.92、2 589.69±856.83及3 250±1 170.44(P<0.05);BC-PLTs血浆分别为3 342.35±1 322.30、3 827.12±1 714.43及3 691.87±1 631.03(P>0.05)。血浆引发fMLP(formyl-Met-Leu-Phe)活化的PMNs呼吸爆发程度(PMNs的引发活性),库存d1、d5 A-PLTs血浆处理组相对于单独加入fMLP阴性对照组其MFI比值相应为1.25±0.1、1.76±0.41(P<0.05)。用小鼠抗人CD154抗体特异性去除血浆中的sCD40L后血浆引发fMLP活化的PMNs呼吸爆发程度相应降低41.84%。以0.1μm滤器滤除血小板微粒后血浆引发fMLP活化的PMNs呼吸爆发程度相应降低69.56%。结论常规库存的血小板随库存时间延长,其血浆中的sCD40L水平升高,并对fMLP活化的PMNs的呼吸爆发具有引发作用。Objective To study the influence of preparation technique and storage time of platelet concentration on sCD40L accumulation and capacity of PLTs free plasma priming PMN respiratory burst.Methods Samples either from APLTs or BC-PLTs which stored for d1,d3 and d5 were collected.Platelet free plasma were isolated by centrifugation Plts at 1 550 g for 20 minutes.sCD40L in plasma were measured by a commercially available ELISA kit.ROS from PMN respiratory burst was measured by oxidation of the indicator dye DHR.The latter had a bright fluorescence which can be detected by flow cytometry.The impact of sCD40L was evaluated by antibody inhibition experiment.Results sCD40L concentrations of A-PLTs plasma（pg / mL） in d1,d3 and d5 were 1 341.62 ± 279.92,2 589.69 ± 856.83,and 3 250 ± 1 170.44（P 0.05）;while those of BC-PLTs plasma（pg / mL） were 3 342.35 ± 1 322.3,3 827.12 ± 1 714.43,and 3 691.87 ± 1 631.03（P 0.05）.The priming activity of A-PLTs plasma to fMLP actived PMNs compared d1 to d5 were（1.25 ± 0.1）,（1.76 ±0.41）（P 0.05）.PMNs priming activity of plasma inhibited by mouse anti-human CD154 monoclonal antibody decreased 41.84% and inhibited by filtration with 0.1 μm membrane decreased 69.56%.Conclusion Amounts of sCD40L accumulation during storage were dependent on preparation technique and storage duration,and was associated with PMNs priming activity.

关 键 词：血小板 库存时间 SCD40L 乏血小板血浆 中性粒细胞 呼吸爆发 

分 类 号：R446.62[医药卫生—诊断学] R331.143[医药卫生—临床医学]