羧基化聚苯乙烯微球与血型糖蛋白偶联及其对血型抗体吸收/释放的效应  

Coupling of carboxy polystyrene microspheres with blood group glycoprotein and its absorption/release effect on blood group antibody

作　　者：余忠清[1] 田鑫华 陈庆[3] 余静薇 YU Zhong-Qing;TIAN Xin-Hua;CHEN Qing;YU Jing-Wei(Department of Blood Transfusion,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China)

机构地区：[1]华中科技大学同济医学院附属协和医院输血科,武汉430022 [2]孝感市精神病医院检验科,孝感432000 [3]华中科技大学同济医学院附属协和医院肝胆外科,武汉430022 [4]华中科技大学同济医学院附属同济医院外科细胞室,武汉430030

出　　处：《中国免疫学杂志》2021年第5期582-586,共5页Chinese Journal of Immunology

基　　金：湖北省自然基金项目(2017CFB802)。

摘　　要：目的:探讨ABO血型糖蛋白-聚苯乙烯微球偶联复合物(CLC)对血型抗体吸收/释放的影响,为孕妇及胎儿ABO新生儿溶血病(HDN)自身血液回输和疑难输血提供新方法。方法:采用EDC/NHS活化羧基化聚苯乙烯微球(CPM)羧基,使其与血型糖蛋白氨基(-NH_(2))脱水缩合形成CLC。红外光谱扫描CPM特征峰波,浊度法计算血型糖蛋白利用率,荧光二抗检测标记效率,吸收/释放试验监测CPM生物学活性。结果:傅立叶红外线扫描显示,1 728~1 630 cm^(-1)处呈现-COOH峰波,暗视野镜下,A血型糖蛋白CLC呈蓝绿色荧光,B血型呈红色荧光,血型糖蛋白与CPM偶联成功。偶联前后A血型糖蛋白平均含量由0.387 5降至0.180 8(P<0.01),B血型糖蛋白由0.387 4降至0.148 7(P<0.01),A/和B血型糖蛋白偶联率分别为53.45%和61.60%。IgM抗A吸收前后平均由477.87降至55.47(P<0.01),IgM抗B平均由460.80降至53.33(P<0.01),56℃放散液中含丰富的IgM抗A/和抗B,与试剂红细胞发生凝集反应(+),提示IgM抗A/和抗B呈可逆性结合。IgG抗A吸收前后效价平均由750.90降至115.20(P<0.01),IgG抗B由537.60降至99.60(P<0.01)。Rh抗D和自身抗体均不被CLC吸收。结论:可溶性血型糖蛋白可与CPM偶联形成CLC,具有良好的免疫吸附与释放功能。Objective:To explore absorption/release effect of ABO blood group glycoprotein and polystyrene microspheres coupling(CLC) complex on born blood group antibody, and to provide a new method for self-transfusion and difficult transfusion of ABO hemolytic disease of new born(HDN) in pregnant women and fetuses.Methods:Carboxyzed polystyrene microspheres(CPM) were activated by EDC/NHS to dehydrate and condense with blood glycoprotein amino group(-NH_(2)) to form blood CLC.Characteristic peak wave of CPM was scanned by infrared spectrum, utilization ratio of blood glycoprotein was calculated by turbidimetry, labeling efficiency was detected by fluorescent second antibody, and biological activity of CPM was monitored by absorption/release test.Results:Fourier infrared scanning showed-COOH peak wave at 1 728~1 630 cm^(-1).Under dark field microscope, A blood group glycoprotein CLC showed blue-green fluorescence, B blood group showed red fluorescence, which suggested that blood glycoprotein and CPM were successfully coupled.Before and after coupling, content of A blood group glycoprotein decreased from 0.387 5 to 0.180 8(P<0.01),B blood group glycoprotein decreased from 0.387 5 to 0.148 7(P<0.01),coupling rate of A/B blood group glycoprotein were 53.45% and 61.60%,respectively.Before and after absorption of IgM anti-A,average value decreased from 477.87 to 55.47(P<0.01).IgM anti-B decreased from 460.80 to 53.33(P<0.01).Before and after the absorption of IgG anti-A,the average titer decreased from 750.90 to 115.20(P<0.01);IgG anti-B decreased from 537.60 to 99.60(P<0.01).Average value decreased from 477.87 to 55.47. Average anti-B value of IgM decreased from 460.80 to 53.33(P<0.01). Dispersion liquid contains IgM anti-A or/B, which agglutinated with standard A or B erythrocyte(+), which suggested that anti-A/or anti-B IgM can bind reversibly.RH anti-D and autoantibodies were not absorbed by CLC.Conclusion:Soluble blood group glycoprotein can be coupled with CPM to form CLC,which has good function of immune-adsorption and re

关 键 词：羧基化聚苯乙烯 血型糖蛋白 脱水缩合 偶联复合物 吸收/释放 

分 类 号：R556.6[医药卫生—血液循环系统疾病]