Epstein-Barr virus reactivation by persistent apical periodontal pathogens
Epstein-Barr virus reactivation by persistent apical periodontal pathogens
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DOI：
10.1111/iej.13255
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发表时间：
2019-12-15
影响因子：
5
通讯作者：
Ogiso, B.
中科院分区：
文献类型：
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作者：
Himi, K.;Takeichi, O.;Ogiso, B.
目的 使用体外和离体方法评估 Epstein-Barr 病毒 (EBV) 重新激活是否由持续性根尖周炎相关微生物触发。方法 对手术切除的人根尖肉芽肿 (n = 50) 和健康牙龈组织 (n = 10) 进行分析，以确定 EBV 和七种持续性根尖周炎相关微生物的存在。此外，采用实时聚合酶链式反应检测EBV即早期基因BZLF-1 mRNA的表达量。还使用三色免疫荧光染色检查了潜膜蛋白 (LMP)-1 和 ZEBRA（由 BZLF-1 编码的 EBV 早期裂解蛋白）的表达。对微生物产生的正丁酸进行了定量，并结合细菌裂解物进行了荧光素酶测定。另外，用细菌裂解物培养Daudi细胞，测定BZLF-1 mRNA和ZEBRA蛋白的表达水平。结果 50 例根尖肉芽肿中有 47 例检测到 EBV DNA 和 BZLF-1 mRNA，但在健康牙龈组织中未检测到。根尖肉芽肿中EBV DNA拷贝数、具核梭杆菌数量与BZLF-1表达量呈显着正相关。中间普雷沃氏菌的数量与BZLF-1的表达量略有相关；然而，其他微生物则不然。根尖肉芽肿中的 CD79a 阳性 B 细胞表达 LMP-1 和 ZEBRA，而健康牙龈组织中的 CD79a 阳性 B 细胞则不然。正丁酸产量在 F. nucleatum 中最高，在 P. intermedia 中最低。粪肠球菌、白色念珠菌和其他测试微生物不产生正丁酸。 F. nucleatum 裂解物以与商业丁酸相同的方式表现出显着增加的 BZLF-1-荧光素酶活性，而 P. intermedia 则没有。 F. nucleatum还诱导Daudi细胞表达BZLF-1 mRNA和ZEBRA蛋白，表明EBV重新激活被诱导。结论 在所测试的持续性根尖周炎相关细菌中，具核梭菌对潜伏 EBV 的再激活作用最强，而粪肠球菌和白色念珠菌以及其他微生物则不然。
Aim To assess whether Epstein-Barr virus (EBV) reactivation is triggered by persistent apical periodontitis-related microbes using in vitro and ex vivo methodologies. Methodology Surgically removed human periapical granulomas (n = 50) and healthy gingival tissues (n = 10) were analysed to determine the presence of EBV and seven persistent apical periodontitis-related microbes. In addition, real-time polymerase chain reaction was used to detect the mRNA expression of BZLF-1, an immediate-early gene of EBV. Expression of latent membrane protein (LMP)-1 and ZEBRA, an early lytic protein of EBV encoded by BZLF-1, was also examined using triple-colour immunofluorescence staining. n-Butyric acid produced by the microbes was quantified, and luciferase assays were performed in association with bacterial lysates. In addition, Daudi cells were cultured with bacterial lysates, and the expression levels of BZLF-1 mRNA and ZEBRA protein were determined. Results EBV DNA and BZLF-1 mRNA were detected in 47 out of 50 periapical granulomas, but not in healthy gingival tissues. The EBV DNA copy number and the number of Fusobacterium nucleatum were significantly positively correlated with BZLF-1 expression in periapical granulomas. The number of Prevotella intermedia was slightly correlated with BZLF-1 expression; however, the other microbes were not. CD79a-positive B cells in periapical granulomas, but not those in healthy gingival tissues, expressed both LMP-1 and ZEBRA. n-Butyric acid production was the highest in F. nucleatum and the lowest in P. intermedia. Enterococcus faecalis, Candida albicans and the other tested microbes did not produce n-butyric acid. An F. nucleatum lysate exhibited significantly increased BZLF-1-luciferase activity in the same manner of commercial butyric acid, whereas P. intermedia did not. F. nucleatum also induced the expression of BZLF-1 mRNA and ZEBRA protein by Daudi cells, indicating that EBV reactivation was induced. Conclusion Among the persistent apical periodontitis-related bacteria that were tested, F. nucleatum most strongly reactivated latent EBV, whereas E. faecalis and C. albicans as well as the other microbes did not.