Höpken Merle, Förster Isabel, Maune Steffen, Brockmann Michael, Schildgen Oliver, Schildgen Verena
Klinik für Hals-, Nasen- und Ohrenheilkunde, Kliniken der Stadt Köln, Klinikum der Privaten Universität Witten/Herdecke mit Sitz in Köln, Cologne, Germany.
Institut für Pathologie, Kliniken der Stadt Köln, Klinikum der Privaten Universität Witten/Herdecke mit Sitz in Köln, Cologne, Germany.
Front Microbiol. 2018 Oct 16;9:2450. doi: 10.3389/fmicb.2018.02450. eCollection 2018.
The human bocavirus (HBoV) is known to persist latently in the infected host cells and seems to replicate its DNA via the DNA damage response system, which is frequently defect in tumors and correlates with microsatellite instability (MSI). Because HBoV is able to persist in the infected tissues, induces pro-fibrotic and pro- cancerogenic cytokines and , and is detected in colorectal and lung tumors, the virus may be involved in cancerogenesis at least as a cofactor. Recently it was shown that the adenotonsillar tissue is an important site of HBoV1 persistence and replication. Considering the background that approximately 60% of oropharyngeal cancers were thought to be attributable to a HPV infection, a co-participation of HBoV in terms of a chronic virus infection might play a role in the cancerogenesis of tonsil tumors. Formalin-fixed, paraffin-embedded tonsil tumor samples were screened for HBoV and HPV DNA. Positive tissue sections were afterward subjected to fluorescence hybridization (FISH) analysis to identify HBoV and HPV infected cells. By use of an cell culture model with primary tonsil fibroblasts, keratinocytes, and lymphocytes infected by HBoV we tried to find the target cells of virus replication. MSI testing was based on a previously published protocol using a de-multiplexed PCR followed by fluorescent detection of PCR products in a capillary sequencing device. In total 62 of 103 (60, 19%) of the tonsil squamous cell carcinomas tested positive for HBoV DNA and 66 of 103 (66%) samples were identified as HPV positive. The FISH analysis revealed both double infection of HPV and HBoV in the same cells as well as single infections of both viruses within the tumor tissue. Twenty-two of 62 HBoV positive tumors tested HPV negative, 40 of 62 tissue sections were HBoV and HPV positive. We analyzed 21 out of the 62 HBoV positive tumors for MSI. Of those four tonsils displayed MSI in at least 1 of 10 microsatellite markers. Our findings support the hypothesis that human bocavirus infections as a cofactor may have an impact on tumor development in tonsils, although it still remains possible that HBoV solely displays a tumor tropism.
人博卡病毒（HBoV）已知可在受感染的宿主细胞中潜伏，并似乎通过DNA损伤反应系统复制其DNA，而该系统在肿瘤中常常存在缺陷且与微卫星不稳定性（MSI）相关。由于HBoV能够在受感染组织中持续存在，诱导促纤维化和促致癌细胞因子，并且在结直肠癌和肺癌肿瘤中被检测到，该病毒可能至少作为辅助因子参与肿瘤发生。最近有研究表明，腺样体扁桃体组织是HBoV1持续存在和复制的重要部位。考虑到约60%的口咽癌被认为归因于HPV感染这一背景，HBoV作为一种慢性病毒感染共同参与可能在扁桃体肿瘤的发生中起作用。对福尔马林固定、石蜡包埋的扁桃体肿瘤样本进行HBoV和HPV DNA筛查。之后，对阳性组织切片进行荧光原位杂交（FISH）分析，以鉴定HBoV和HPV感染的细胞。通过使用由HBoV感染的原代扁桃体成纤维细胞、角质形成细胞和淋巴细胞组成的细胞培养模型，我们试图找到病毒复制的靶细胞。MSI检测基于先前发表的方案，使用多重PCR，随后在毛细管测序装置中对PCR产物进行荧光检测。在总共103例扁桃体鳞状细胞癌中，62例（60.19%）HBoV DNA检测呈阳性，103例样本中有66例（66%）被鉴定为HPV阳性。FISH分析显示，在同一细胞中HPV和HBoV存在双重感染，以及在肿瘤组织中两种病毒均存在单一感染。62例HBoV阳性肿瘤中有22例HPV检测呈阴性，62例组织切片中有40例HBoV和HPV均呈阳性。我们对62例HBoV阳性肿瘤中的21例进行了MSI分析。其中4例扁桃体在10个微卫星标记中的至少1个显示出MSI。我们的研究结果支持这样的假设，即人博卡病毒感染作为辅助因子可能对扁桃体肿瘤的发展产生影响，尽管HBoV仍有可能仅表现出肿瘤嗜性。
