人血小板表面CD36糖蛋白检测方法的建立及初步应用

Establishment of Method Detecting CD36 Expression on Human Platelet and Its Application

个体血小板表面CD36抗原缺乏在随机输注时有产生抗-CD36免疫反应的风险,是血小板输注无效的原因之一。本研究应用流式细胞术检测杭州地区单采血小板供者的血小板表面CD36抗原表达情况,并分析个体血小板上CD36缺失表型的频率。留取献血者新鲜抗凝血样,经离心获取富血小板血浆,洗涤并调整血小板计数至1×106。采用CD36-FITC、CD41-PE单克隆抗体和血小板孵育反应,然后用流式细胞仪检测和分析血小板表面糖蛋白CD36抗原表达情况。对于血小板表面CD36抗原阴性的标本,进一步筛查其单核细胞表面CD36的表达情况。结果表明:192例无偿献血者筛查出7例血小板表面CD36抗原阴性,CD36缺失型频率为3.6%,均为Ⅱ型缺失。人群中个体CD36抗原表达强度存在差异,参照CD36几何平均荧光强度数值大小,59例为低表达,126例为高表达。结论:人群中存在CD36Ⅱ型缺失表型,这些数据将为研究CD36抗原分布提供参考,有助于解决血小板输注无效问题。

The individual with the deficiency of CD36 antigen on platelet displayed the risk of anti-CD36 immune reaction induced by transfusion,which is one of the reasons for platelet transfusion refractoriness（PTR）.This study was purposed to detect the expression level of CD36 antigen on platelet by flow cytometry among aphoresis platelet donors of Hangzhou area,and the frequency of CD36 deficiency was analyzed.Platelet-rich plasma（PRP） was separated from fresh anticoagulant whole blood by centrifugation,then the platelets were washed and adjusted to 1×10^6.The platelets were incubated with FITC-labeled CD36 and PE-labeled CD41 monoclonal antibodies,then the expression level of CD36 was detected by flow cytometry.The CD36 expression on monocytes for the samples of CD36-deficiency on the platelets was further analyzed.The results showed that 7 samples with CD36 antigen deficiency were found in 192 aphoresis platelet donors.The frequency of CD36 deficiency was 3.6% and all of them were typeⅡ deficiency.The significant difference of CD36 antigen expression was observed in the platelet donors of Hangzhou population,among them 59 individuals with low expressed CD36 antigen and 126 individuals with highly expressed CD36 antigen were found according to the geometric mean fluorescence intensity.It is concluded that the CD36 antigen deficient phenotype existed in the population,these data will provide the information for research of the CD36 antigen distribution and help to solve the platelet transfusion refractoriness.