Shimakage Misuzu, Sasagawa Toshiyuki, Kimura Michio, Shimakage Tatsuya, Seto Shiro, Kodama Ken, Sakamoto Haruhiko
Clinical Research Institute, Osaka National Hospital, Osaka, Japan.
Hum Pathol. 2004 Jul;35(7):862-8. doi: 10.1016/j.humpath.2004.03.017.
Langerhans' cell histiocytosis (LCH) is a proliferative histiocytic disorder of unknown etiology. We previously reported that Epstein-Barr virus (EBV) infects and proliferates in macrophages, and investigated the possibility that EBV exhibits etiologic effects in LCH. To detect EBV expression, paraffin sections from 17 LCH cases were examined by mRNA in situ hybridization for EBV BamHIW, Epstein-Barr virus nuclear antigen-2 (EBNA2), and Epstein-Barr virus-encoded small nonpolyadenylated RNA (EBER1) sequences, and by indirect immunofluorescence staining for EBNA2, latent membrane protein 1 (LMP1), and BamHIZ-coding leftward-reading frame 1 (BZLF1). To detect EBV DNA, polymerase chain reaction (PCR)-Southern blotting was used. All cases showed positive hybridization signals by BamHIW mRNA in situ hybridization. Also, 13 and 14 cases showed positive signals for EBNA2 and EBER1 RNA in situ hybridization, respectively. Furthermore, almost all cases exhibited fluorescence after immunofluorescence staining with monoclonal anti-EBNA2 and anti-BZLF1 antibodies, and 15 cases were positive after treatment with monoclonal anti-LMP1 antibody. PCR-Southern blotting detected an amplified EBER1 sequence in all 9 cases examined. EBV expression was confirmed in LCH using in situ hybridization and immunofluorescence. Furthermore, EBV DNA was also detected by PCR-Southern blotting. These positive results of BZLF1 suggest that EBV replicates in LCH tissues.
朗格汉斯细胞组织细胞增多症（LCH）是一种病因不明的组织细胞增生性疾病。我们之前报道过，爱泼斯坦-巴尔病毒（EBV）可在巨噬细胞中感染和增殖，并研究了EBV在LCH中发挥病因学作用的可能性。为检测EBV表达，对17例LCH病例的石蜡切片进行了EBV BamHIW、爱泼斯坦-巴尔病毒核抗原-2（EBNA2）和爱泼斯坦-巴尔病毒编码的小非聚腺苷酸化RNA（EBER1）序列的mRNA原位杂交检测，以及对EBNA2、潜伏膜蛋白1（LMP1）和BamHIZ编码的左向阅读框1（BZLF1）的间接免疫荧光染色检测。为检测EBV DNA，采用了聚合酶链反应（PCR）-Southern印迹法。所有病例通过BamHIW mRNA原位杂交均显示阳性杂交信号。此外，分别有13例和14例病例在EBNA2和EBER1 RNA原位杂交中显示阳性信号。此外，几乎所有病例在用抗EBNA2和抗BZLF1单克隆抗体进行免疫荧光染色后均呈现荧光，15例在用抗LMP1单克隆抗体处理后呈阳性。PCR-Southern印迹法在所有检测的9例病例中均检测到扩增的EBER1序列。通过原位杂交和免疫荧光证实了LCH中EBV的表达。此外，通过PCR-Southern印迹法也检测到了EBV DNA。BZLF1的这些阳性结果表明EBV在LCH组织中复制。
