抗-CD36抗体引起的血小板输注无效及诊疗策略

Ineffective platelet transfusion caused by anti-CD36 antibody and the effective treatment strategy

目的探讨CD36基因缺失表达患者产生抗-CD36抗体引起的血小板输注无效(PTR)及配型效果追踪。方法用流式细胞仪检测患者血小板和单核细胞上CD36抗原的表达,PAKPLUS试剂盒检测患者血清中血小板膜糖蛋白(包括GPIIb/IIIa、GP Ib/IX、GP Ia/IIa、GPIV)和HLA抗体,从患者全血中提取DNA,通过PCR-SBT方法检测CD36基因突变。通过CD36阴性表达供者库,挑选合适供者,进行血小板配型,血小板配型使用自制的固相凝集试剂盒。结果流式细胞仪检测出患者血小板和单核细胞上CD36表达均为阴性,确定为I型CD36缺失型。从患者血浆中检出抗-GPIV(即CD36)抗体,通过测序发现患者CD36基因有如下突变:exon5 delAC329-330、exon12C1156T、exon14 C1409T(均为杂合突变)。患者输注相合供者的配型血小板后,24 h血小板计数由输注前3×10^9/L上升至55×10^9/L。结论除了HLA抗原引起的免疫致敏外,CD36抗原引起的PTR也应引起重视,对于CD36缺失表达的血小板输注无效患者而言,必须输注CD36阴性血小板才是有效的治疗措施。

Objective To investigate platelet transfusion ineffectiveness(PTR)caused by anti-CD36 antibody production in patients with CD36 gene deletion and evaluate the management of cross-matchin transfusion.Methods Flow cytometry was used to detect the expression of CD36 antigen on platelets and monocytes of patients.PAKPLUS kit was used to detect platelet membrane glycoproteins(including GPIIb/IIIa,GP Ib/IX,GP Ia/IIa,GPIV)and HLA antibodies in patients′serum.DNA was extracted from patients′whole blood,and CD36 gene mutation was detected by PCR-SBT method.The CD36 negative expression donor library was used to select suitable donors for platelet matching.Platelet matching was performed using a selfmade solid phase agglutination kit.Results The patient was detected as type I CD36 deletion showing no CD36 expression on platelets and monocytes in flow cytometry analysis.Anti-CD36 antibodies were detected from the patient′s plasma,and the patient′s CD36 gene was found to have the following mutations by sequencing:exon5 delAC329-330,exon12 C1156 T,exon14 C1409 T(all heterozygous mutations).After transfusion of matched donor platelets from the donor,the platelet count at24 hours increased from 3×10^9/L before the transfusion to 55×10^9/L.Conclusion In addition to the immune sensitization caused by the HLA antigen,the PTR caused by the CD36 antigen should also be paid attention to.For patients with CD36 expression deficiency should receive CD36 negative platelets transfusion.