Baumgarten Peter, Michaelis Martin, Rothweiler Florian, Starzetz Tatjana, Rabenau Holger F, Berger Annemarie, Jennewein Lukas, Braczynski Anne K, Franz Kea, Seifert Volker, Steinbach Joachim P, Allwinn Regina, Mittelbronn Michel, Cinatl Jindrich
Neurological Institute (Edinger Institute), Goethe University, Frankfurt am Main, Germany (P.B., T.S., L.J., A.K.B., Mi.M.); Institute of Medical Virology, Goethe University, Frankfurt am Main, Germany (Ma.M., F.R., H.F.R., A.B., R.A., J.C.); German Cancer Consortium, Heidelberg, Germany (J.P.S., Mi.M.); German Cancer Research Center, Heidelberg, Germany (J.P.S., Mi.M.); Department of Neurosurgery, Goethe University, Frankfurt am Main, Germany (K.F., V.S.); Senckenberg Institute of Neurooncology, University of Frankfurt am Main, Germany (K.F., J.P.S.).
Neuro Oncol. 2014 Nov;16(11):1469-77. doi: 10.1093/neuonc/nou167. Epub 2014 Aug 25.
Experimental findings have suggested that human cytomegalovirus (HCMV) infection of tumor cells may exert oncomodulatory effects that enhance tumor malignancy. However, controversial findings have been published on the presence of HCMV in malignant tumors. Here, we present the first study that systematically investigates HCMV infection in human nervous system tumors by highly sensitive immunohistochemistry in correlation with the HCMV serostatus of the patients.
Immunohistochemical and quantitative PCR-based methods to detect different HCMV antigens and genomic HCMV DNA were optimized prior to the investigation of pathological samples. Moreover, the pathological results were matched with the HCMV serostatus of the patients.
HCMV immediate-early, late, and pp65 antigens could be detected in single cells from HCMV strain Hi91-infected UKF-NB-4 neuroblastoma cells after 1:1024 dilution with noninfected UKF-NB-4 cells. Genomic HCMV DNA could be detected in copy numbers as low as 430 copies/mL. However, we did not detect HCMV in tumors from a cohort of 123 glioblastoma, medulloblastoma, or neuroblastoma patients. Notably, we detected nonspecifically positive staining in tumor tissues of HCMV seropositive and seronegative glioblastoma patients. The HCMV serostatus of 67 glioblastoma patients matched the general epidemiological prevalence data for Western countries (72% of female and 57% of male glioblastoma patients were HCMV seropositive). Median survival was not significantly different in HCMV seropositive versus seronegative glioblastoma patients.
The prevalence of HCMV-infected tumor cells may be much lower than previously reported based on highly sensitive detection methods.
实验结果表明，肿瘤细胞的人巨细胞病毒（HCMV）感染可能发挥促癌作用，增强肿瘤恶性程度。然而，关于恶性肿瘤中HCMV的存在，已有相互矛盾的研究结果发表。在此，我们首次通过高灵敏度免疫组织化学方法，结合患者的HCMV血清学状态，系统地研究了人神经系统肿瘤中的HCMV感染情况。
在对病理样本进行研究之前，优化了基于免疫组织化学和定量PCR的方法，以检测不同的HCMV抗原和基因组HCMV DNA。此外，将病理结果与患者的HCMV血清学状态进行匹配。
用未感染的UKF-NB-4细胞以1:1024稀释HCMV毒株Hi91感染的UKF-NB-4神经母细胞瘤细胞后，可在单细胞中检测到HCMV即刻早期、晚期和pp65抗原。基因组HCMV DNA的检测拷贝数低至430拷贝/毫升。然而，我们在123例胶质母细胞瘤、髓母细胞瘤或神经母细胞瘤患者的肿瘤中未检测到HCMV。值得注意的是，我们在HCMV血清阳性和血清阴性胶质母细胞瘤患者的肿瘤组织中检测到非特异性阳性染色。67例胶质母细胞瘤患者的HCMV血清学状态与西方国家的一般流行病学患病率数据相符（72%的女性和57%的男性胶质母细胞瘤患者HCMV血清阳性）。HCMV血清阳性与血清阴性胶质母细胞瘤患者的中位生存期无显著差异。
基于高灵敏度检测方法，HCMV感染肿瘤细胞的患病率可能远低于先前报道。
