Takeda T, Mizugaki Y, Matsubara L, Imai S, Koike T, Takada K
Hokkaido University School of Medicine, Sapporo, Japan.
Arthritis Rheum. 2000 Jun;43(6):1218-25. doi: 10.1002/1529-0131(200006)43:6<1218::AID-ANR4>3.0.CO;2-2.
To evaluate the existence of Epstein-Barr virus (EBV) infection in the synovial tissue of patients with rheumatoid arthritis (RA).
Synovial tissues were obtained at synovectomy or arthroplasty from 32 patients with RA and 30 control patients with osteoarthritis (OA). EBV DNA was detected by Southern blot hybridization and/or polymerase chain reaction (PCR) amplification. To localize the EBV-infected cells, tissue sections were studied by RNA in situ hybridization (ISH) for the EBV-encoded small RNA 1 (EBER-1), by DNA ISH for the Bam HI W region of EBV DNA, and by immunohistochemistry for EBV lytic proteins BZLF1 and gp350/220.
EBV DNA was detected by PCR in 15 of the 32 samples from RA patients (47%), but in none of those from the 30 OA patients (P < 0.01). Of the 15 PCR-positive samples, 9 contained >1 EBV copy/1,000 cells (referred to as EBV 2+), and 6 contained 1 copy/1,000-5,000 cells (EBV 1+). Among the 9 EBV 2+ samples, 3 were also positive for EBV DNA by Southern blot hybridization, 5 were positive for EBER-1 by RNA ISH, and 3 were positive for EBV DNA by DNA ISH. Immunohistochemical analysis showed positive signals in all samples for BZLF1 and in 7 samples for gp350/ 220. In each examination, the positive signals were detected not only in lymphocytes, but also in synovial lining cells.
EBV was frequently detected in the synovial tissue of RA patients. The infected cells were both lymphocytes and synovial cells, and expressed EBV proteins associated with virus replication. These findings suggest that EBV may play a role in the pathogenesis of RA.
评估类风湿关节炎（RA）患者滑膜组织中是否存在EB病毒（EBV）感染。
在滑膜切除术或关节成形术中获取32例RA患者和30例骨关节炎（OA）对照患者的滑膜组织。通过Southern印迹杂交和/或聚合酶链反应（PCR）扩增检测EBV DNA。为了定位EBV感染的细胞，通过RNA原位杂交（ISH）检测EBV编码的小RNA 1（EBER-1）、通过DNA ISH检测EBV DNA的Bam HI W区域、以及通过免疫组织化学检测EBV裂解蛋白BZLF1和gp350/220，对组织切片进行研究。
通过PCR在32例RA患者的15份样本中检测到EBV DNA（47%），但在30例OA患者的样本中均未检测到（P < 0.01）。在15份PCR阳性样本中，9份含有>1个EBV拷贝/1000个细胞（称为EBV 2+），6份含有1个拷贝/1000 - 5000个细胞（EBV 1+）。在9份EBV 2+样本中，3份通过Southern印迹杂交检测EBV DNA也呈阳性，5份通过RNA ISH检测EBER-1呈阳性，3份通过DNA ISH检测EBV DNA呈阳性。免疫组织化学分析显示，所有样本中BZLF1均呈阳性信号，7份样本中gp350/220呈阳性信号。在每次检测中，阳性信号不仅在淋巴细胞中检测到，也在滑膜衬里细胞中检测到。
在RA患者的滑膜组织中经常检测到EBV。感染的细胞既有淋巴细胞也有滑膜细胞，并表达与病毒复制相关的EBV蛋白。这些发现表明EBV可能在RA的发病机制中起作用。
