Elsaid Dina Samir, Elbedewy Tamer Abd Elhamid, Soliman Nema Ali, Shalaby Kamal Ali, Haroun Riham Abdel-Hamid
Department of Biochemistry, Faculty of Science, Ain Shams University, Cairo, Egypt.
Department of Internal Medicine, Faculty of Medicine, Tanta University, Tanta, Egypt.
Int J Lab Hematol. 2025 Apr;47(2):255-265. doi: 10.1111/ijlh.14391. Epub 2024 Nov 6.
Immune thrombocytopenic purpura (ITP) is an acquired autoimmune disease characterized by reduced platelet counts due to immune system dysregulation caused by many factors, including genetics, autoimmune diseases, infections, and inflammations. Therefore, the current study aimed to evaluate immunological markers such as the expression level of lysosomal associated membrane protein 3 (LAMP-3), also known as CD63, and the expression level of Fc-gamma receptor I (FcγRI), also known as CD64 and also investigate the association of Fc-gamma receptor IIIA (FcγRIIIA) 158 V/F polymorphism to the risk of ITP.
A total of 180 subjects; 60 ITP patients, 60 patients with thrombocytopenia of other causes and 60 controls were enrolled into our study. The expression level of CD63 was done using reverse transcription quantitative PCR (RTqPCR), while CD64 expression level was done by flow cytometry. The polymorphism of FcγRIIIA 158 V/F gene was analyzed by polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) analysis. Finally, CD63 and CD64 protein-protein interactions were done by using the STRING online database.
The expression of CD63 was significantly elevated in ITP patients than thrombocytopenia patients and healthy control. Also there was high expression level of CD64 on granulocytes and monocytes from ITP patients than other groups. Receiver operating characteristic curve (ROC curve) analysis of CD63 showed an area under the curve (AUC) revealed of 1.00, sensitivity of 100% and specificity of 100%; while for CD64 on granulocytes, AUC of 0.998 as well as a sensitivity of 96.66% and specificity of 93.33%. Regarding FcγRIIIa 158 V/F polymorphism, all patients and healthy volunteers included in this study showed the wild FF genotype.
The expression of both CD63 and CD64 were significantly increased in ITP patients and could be good biomarkers to diagnose ITP. Additionally, there is no association between FcγRIIIa 158 V/F polymorphism and the risk of ITP disease.
免疫性血小板减少性紫癜(ITP)是一种获得性自身免疫性疾病,其特征是由于多种因素(包括遗传、自身免疫性疾病、感染和炎症)导致免疫系统失调而使血小板计数减少。因此,本研究旨在评估免疫标志物,如溶酶体相关膜蛋白3(LAMP-3,也称为CD63)的表达水平以及Fc-γ受体I(FcγRI,也称为CD64)的表达水平,并研究Fc-γ受体IIIA(FcγRIIIA)158V/F多态性与ITP风险的相关性。
本研究共纳入180名受试者,包括60例ITP患者、60例其他原因导致血小板减少的患者和60名对照。采用逆转录定量PCR(RTqPCR)检测CD63的表达水平,通过流式细胞术检测CD64的表达水平。采用聚合酶链反应(PCR)及限制性片段长度多态性(PCR-RFLP)分析检测FcγRIIIA 158V/F基因多态性。最后,利用STRING在线数据库分析CD63和CD64的蛋白质-蛋白质相互作用。
ITP患者中CD63的表达明显高于血小板减少患者和健康对照。ITP患者粒细胞和单核细胞上CD64的表达水平也高于其他组。CD63的受试者工作特征曲线(ROC曲线)分析显示曲线下面积(AUC)为1.00,敏感性为100%,特异性为100%;而粒细胞上CD64的AUC为0.998,敏感性为96.66%,特异性为93.33%。关于FcγRIIIa 158V/F多态性,本研究纳入的所有患者和健康志愿者均表现为野生型FF基因型。
ITP患者中CD63和CD64的表达均显著增加,可能是诊断ITP的良好生物标志物。此外,FcγRIIIa 158V/F多态性与ITP疾病风险之间无相关性。