Egawa H, Ohishi T, Arai T, Inomata Y, Uemoto S, Asonuma K, Kinchi T, Okajima H, Matsui A, Kawashima N, Martinez O M, Tanaka K
Department of Transplanation Immunology, Faculty of Medicine, Kyoto University, Japan.
Clin Transplant. 1998 Apr;12(2):116-22.
For quantitative assessment of ongoing symptomatic Epstein-Barr virus (EBV) infection in pediatric recipients of liver transplantation, we determined the number of peripheral blood mononuclear cells (PBMC) infected by EBV by in situ hybridization (ISH) and related the results with clinical courses of those patients. Twenty-four patients had symptomatic EBV infection between February 1995 and March 1996. Blood samples were obtained from these 24 patients at the time of acute phase, from 13 of them during convalescence, and 37 pediatric patients before transplantation. ISH was performed on the PBMC and polymerase chain reaction (PCR) on DNA from whole blood. Oligonucleotide probes for ISH were chosen from coding sequences of EBV-encoded small nuclear RNA 1 (EBER1). Results of ISH were reported in a number of cells expressing EBER1/5 x 104 PBMC (#EBER1). Fever, diarrhea, upper respiratory symptoms, pleural effusion, ascites, lymphadenopathy, and lymphoproliferative disease (LPD) accompanied with EBV infection proven by serology, viral-specific stain or PCR were regarded as EBV related diseases (EBVD). All samples with positive #EBER1 were accompanied by positive EBV PCR. #EBERI was 68.2 +/- 144.9 (mean +/- SD) ranging from 0 to 621 in the acute phase, 0.20 +/- 0.41 ranging from 0 to 2 in the convalescence phase, 0.27 +/- 0.77 in 23 preoperative patients with positive serology, and 0 in all 14 preoperative patients with negative serology. The #EBER1 in ongoing EBVD was significantly greater than that of patients in convalescence or before transplantation. Patients with #EBERI greater than 10 had a significantly lower chance of convalescence and a higher mortality than patients with #EBER 1 less than 10. We conclude that #EBER1 could be a specific and quantitative marker of EBVD and might predict progression to LPD.
为了对肝移植儿科受者中正在发生的有症状的爱泼斯坦-巴尔病毒（EBV）感染进行定量评估，我们通过原位杂交（ISH）确定了受EBV感染的外周血单个核细胞（PBMC）的数量，并将结果与这些患者的临床病程相关联。1995年2月至1996年3月期间，24例患者发生了有症状的EBV感染。在急性期从这24例患者采集血样，在恢复期从其中13例采集血样，并在移植前从37例儿科患者采集血样。对PBMC进行ISH检测，对全血DNA进行聚合酶链反应（PCR）。ISH的寡核苷酸探针选自EBV编码的小核RNA 1（EBER1）的编码序列。ISH结果以表达EBER1的细胞数/5×10⁴PBMC（#EBER1）报告。发热、腹泻、上呼吸道症状、胸腔积液、腹水、淋巴结病以及经血清学、病毒特异性染色或PCR证实伴有EBV感染的淋巴增殖性疾病（LPD）被视为EBV相关疾病（EBVD）。所有#EBER1阳性的样本均伴有EBV PCR阳性。急性期#EBER1为68.2±144.9（平均值±标准差），范围为0至621；恢复期为0.20±0.41，范围为0至2；血清学阳性的23例术前患者为0.27±0.77，血清学阴性的所有14例术前患者为0。正在发生的EBVD中的#EBER1显著高于恢复期或移植前患者。#EBER1大于10的患者康复机会显著低于#EBER1小于10的患者，且死亡率更高。我们得出结论，#EBER1可能是EBVD的特异性定量标志物，并且可能预测向LPD的进展。
