Department of Pediatrics and Developmental Biology, Tokyo Medical and Dental University (TMDU), Tokyo, Japan.
Laboratory of Lymphocyte Activation and Susceptibility to EBV Infection, UMR 1163, INSERM, Imagine Institute, Paris, France.
Int J Hematol. 2024 Aug;120(2):241-251. doi: 10.1007/s12185-024-03786-0. Epub 2024 May 3.
When Epstein-Barr virus (EBV) infection is suspected, identification of infected cells is important to understand the pathogenesis, determinine the treatment strategy, and predict the prognosis. We used the PrimeFlow™ RNA Assay Kit with a probe to detect EBV-encoded small RNAs (EBERs) and multiple surface markers, to identify EBV-infected cells by flow cytometry. We analyzed a total of 24 patients [11 with chronic active EBV disease (CAEBV), 3 with hydroa vacciniforme lymphoproliferative disorder, 2 with X-linked lymphoproliferative disease type 1 (XLP1), 2 with EBV-associated hemophagocytic lymphohistiocytosis, and 6 with posttransplant lymphoproliferative disorder (PTLD)]. We compared infected cells using conventional quantitative PCR methods and confirmed that infected cell types were identical in most patients. Patients with CAEBV had widespread infection in T and NK cells, but a small amount of B cells were also infected, and infection in patients with XLP1 and PTLD was not limited to B cells. EBV-associated diseases are believed to be complex pathologies caused by EBV infecting a variety of cells other than B cells. We also demonstrated that infected cells were positive for HLA-DR in patients with CAEBV. EBER flow FISH can identify EBV-infected cells with high sensitivity and is useful for elucidating the pathogenesis.
当怀疑 EBV 感染时，鉴定感染细胞对于了解发病机制、确定治疗策略和预测预后非常重要。我们使用了带有 EBV 编码小 RNA（EBERs）和多个表面标志物探针的 PrimeFlow™ RNA 检测试剂盒，通过流式细胞术鉴定 EBV 感染细胞。我们共分析了 24 例患者[11 例慢性活动性 EBV 疾病（CAEBV）、3 例水疱疹样淋巴组织增生性疾病、2 例 X 连锁淋巴组织增生性疾病 1 型（XLP1）、2 例 EBV 相关噬血细胞性淋巴组织细胞增生症和 6 例移植后淋巴组织增生性疾病（PTLD）]。我们使用传统定量 PCR 方法比较了感染细胞，并证实大多数患者的感染细胞类型相同。CAEBV 患者的 T 和 NK 细胞广泛感染，但也有少量 B 细胞被感染，而 XLP1 和 PTLD 患者的感染并不仅限于 B 细胞。人们认为 EBV 相关疾病是由 EBV 感染除 B 细胞以外的多种细胞引起的复杂病理。我们还证明 CAEBV 患者的感染细胞 HLA-DR 阳性。EBER 流式荧光原位杂交（FISH）可以高灵敏度地鉴定 EBV 感染细胞，有助于阐明发病机制。
