造血干细胞移植术后抗-CD36抗体介导的血小板输注无效症和相关病例的实验研究  

Anti-CD36 Mediated Platelet Transfusion Refractoriness and Related Cases After Stem Cell Transplantation

作　　者：周燕 李丽兰 钟周琳 刘学军 刘金莲 申卫东 吴国光 ZHOU Yan, LI Li-Lan, ZHONG Zhou-Lin, LIU Xue-Jun, LIU Jin-Lian, SHEN Wei-Dong, WU Guo-Guang(Nan-ning Institute of Transfusion Medicine, Nanning 530007, Guangxi Zhuang Autonomous Region, Chin)

机构地区：[1]南宁输血医学研究所,广西南宁530007

出　　处：《中国实验血液学杂志》2018年第2期541-546,共6页Journal of Experimental Hematology

基　　金：广西科学研究与技术开发计划项目(08160-05);广西自然科学基金项目(2017GXNSFAA198240);广西自然科学基金项目(2016GXNSFAA380143);南宁市科学研究与技术开发计划科技重大专项(20173117)

摘　　要：目的:对造血干细胞移植术后由抗-CD36抗体介导血小板输注无效症和相关病例进行实验研究,对患者和造血干细胞供者的CD36表型和CD36基因,以及相关抗体的特征进行分析和鉴定,探讨患者的血小板输注治疗效果。方法:采用流式细胞术检测患者及造血干细胞供者CD36在血小板及单核细胞上的表达;采用本实验室建立的快速血小板抗原单克隆抗体特异性免疫固定检测技术(fast monoclonal antibody-specific immobilization of platelet antigen,F-MAIPA)鉴定患者血清中的血小板抗体;采用测序(polymerase chain reaction sequence-based typing,PCRSBT)方法分析患者及供者CD36基因序列及HPA序列;应用STR-PCR(short tandem repeat polymerase chain reaction,STR-PCR)技术监测患者植入的证据。从CD36缺失血小板库中挑选CD36表达缺失的供者血小板给予患者输注,并监测其血小板计数。结果:造血干细胞供者为CD36+,而患者为I型CD36缺失者。患者移植后血清中存在抗-CD36抗体并排除HLA及HPA相关抗体;患者CD36外显子测序提示为1个新的CD36突变基因,即CD36Exon 6-1G>C纯合子(突变位于剪切位点)突变基因所致的CD36缺失。移植后18 d患者的STR位点、HPA以及CD36型别已完全转化为供者型别(完全嵌合),移植后96 d患者血小板、单个核细胞上已有CD36表达。给该患者输注CD36缺失血小板后血小板计数明显提高。结论:对造血干细胞移植术后由抗-CD36抗体引起的输血小板无效症,须对CD36在移植中的同种免疫反应给予足够重视。为了确保血小板输注有效,对CD36缺失患者需给予CD36缺失血小板输注。Objective： To analyse the cases of platelet transfusion refractoriness after received HLA-matched unrelated donor hematopoietic stem cell transplantation, to analyze and identify the phenotype and genotype of CD36 in both the patient and stem cell donor, as well as the characteristic of antibody induced platelet transfusion refractoriness, and to analyse the efficacy of matched CD36-deficiency platelets transfusions. Methods： The CD36 expression on platelet and monocyte was analyzed by flow cytometry （FCM） in both patient and donor. Polymerase chain reaction sequence-based typing （PCR-SBT） was used to analyze the exons sequence of CD36 and HPA. Fast monoclonal antibody-specific immobilization of platelet antigen （F-MAIPA） and FCM were used to identify platelet antibodies in the patient. Short tandem repeat polymerase chain reaction （STR-PCR） was applied to monitor engraftment evidence. The platelet level was monitored. CD36- deficiency donor＇s platelets were selected from CD36- deficiency donor blood bank. Results： The donor was CD36 positive and the patient was typed I CD36 deficiency. The anti-CD36 antibody was identified in patient＇s serum （after transplantation）, while the HLA and HPA-related antibodies were excluded. Sequence analysis of CD36 exon in the patient showed Exon 6 -1G 〉 C（ Change in splicing site）homozygote, which was a novel CD36 mutation. STR, HPA and CD36 of the patient （complete chimerism） were conversed to that of donor gene types on day 18 after allo-HSCT. The positive CD36 expression on platelet and monocyte in the patient was observed on day 96 after allo-HSCT. The patient showed the platelet transfusion refractoriness which was significantly improved after platelets transfusions from CD36 deficiency donors. Conclusion： Stem cell transplants resulted in anti-CD36 and caused platelet transfusion refractoriness, that was first reported in China. To ensure the efficacy of platelet transfusion, the CD36-deficiency patient should receive CD36 deficienc

关 键 词：造血干细胞 造血干细胞移植 抗-CD36抗体 血小板输注无效 

分 类 号：R457.7[医药卫生—治疗学] R558[医药卫生—临床医学]