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TCRVδ亚型表达的流式细胞术模式可快速识别γδT细胞淋巴瘤。

Flow Cytometric Pattern of TCRVδ Subtype Expression Rapidly Identifies γδT Cell Lymphoma.

作者信息

Chen Xiao, Zhao Sishu, Liu Lu, Qiao Chun, Wang Yan, Fan Lei, Jin Huimin, Wu Yujie

机构信息

Department of Hematology, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.

Key Laboratory of Hematology of Nanjing Medical University, Nanjing, China.

出版信息

Front Oncol. 2020 Jun 16;10:844. doi: 10.3389/fonc.2020.00844. eCollection 2020.

Abstract

γδT cell lymphoma (γδ TCL) is a class of hematopoietic malignancy that expresses the γδ T cell receptor (TCR) with a low incidence. Determining the clonal proliferation of γδT cells is important for the diagnosis of such malignancies. Few studies have used flow cytometry to detect VδTCR and its subtypes (Vδ1 and Vδ2) at the protein level, although it is a practical method for determining the neoplastic γδT cells. A TCRVδ-based 10-color protocol was designed for the detection of malignant proliferation of γδT subtype cells by multiparameter flow cytometry, and the diagnostic results were compared with the gene rearrangement results. All 19 cases of γδ TCL were positive for cluster of differentiation 3 (CD3) and TCR γδ and presented with abnormal distribution patterns of Vδ1 and Vδ2, of which 16 of the 19 cases showed a restricted Vδ1 staining pattern and the remaining three cases lacked the expression of either Vδ1 or Vδ2. Among the 10 normal controls and 11 patients with reactively higher CD4 and CD8 double-negative ratio, the percentage of Vδ2 positive events (range: 16.4-99.0%) was significantly higher than that of Vδ1 (range: 0-50.5%; < 0.0001), and all cases had a normal Vδ distribution pattern. To detect clonality, there was no difference in the detection rate between the TCRVδ analysis and the gene scanning techniques ( = 1.000) with a high degree of coincidence (Kappa = 0.850, < 0.001). The heteroduplex analysis was less sensitive than the other methods but was more specific (100%) than the gene scanning techniques, and the TCRVδ subtype analysis had the highest sensitivity, specificity, positive predictive value, and negative predictive value. Compared with molecular methods, immunophenotyping is able to distinguish the T cell lineage. The γδT panel, based on the TCRVδ antibody by flow cytometry, could be advantageous for the rapid identification of suspected γδTCL.

摘要

γδT细胞淋巴瘤(γδTCL)是一类发病率较低的表达γδT细胞受体(TCR)的造血系统恶性肿瘤。确定γδT细胞的克隆增殖对于此类恶性肿瘤的诊断很重要。尽管流式细胞术是确定肿瘤性γδT细胞的一种实用方法,但很少有研究使用该技术在蛋白质水平检测VδTCR及其亚型(Vδ1和Vδ2)。设计了一种基于TCRVδ的10色方案,用于通过多参数流式细胞术检测γδT亚型细胞的恶性增殖,并将诊断结果与基因重排结果进行比较。19例γδTCL患者的分化簇3(CD3)和TCRγδ均呈阳性,且Vδ1和Vδ2呈现异常分布模式,其中19例中有16例表现为Vδ1染色模式受限,其余3例缺乏Vδ1或Vδ2的表达。在10名正常对照者和11例CD4和CD8双阴性比例反应性较高的患者中,Vδ2阳性事件的百分比(范围:16.4% - 99.0%)显著高于Vδ1(范围:0 - 50.5%;P < 0.0001),且所有病例的Vδ分布模式均正常。为检测克隆性,TCRVδ分析与基因扫描技术的检测率无差异(P = 1.000),一致性较高(Kappa = 0.850,P < 0.001)。异源双链分析比其他方法敏感性低,但比基因扫描技术更具特异性(100%),TCRVδ亚型分析具有最高的敏感性、特异性、阳性预测值和阴性预测值。与分子方法相比,免疫表型分析能够区分T细胞谱系。基于流式细胞术TCRVδ抗体的γδT检测板可能有利于快速识别疑似γδTCL。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/001c/7308429/bbd9e2ad5651/fonc-10-00844-g0001.jpg

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