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Quantitative analysis of viral load per haploid genome revealed the different biological features of Merkel cell polyomavirus infection in skin tumor
Authors:Satoshi Ota  Shumpei Ishikawa  Yutaka Takazawa  Akiteru Goto  Takeshi Fujii  Ken-ichi Ohashi  Masashi Fukayama
Institution:Department of Pathology, Chiba University Hospital, University of Chiba, Chuo, Chiba, Chiba, Japan.
Abstract:Merkel cell polyomavirus (MCPyV) has recently been identified in Merkel cell carcinoma (MCC), an aggressive cancer that occurs in sun-exposed skin. Conventional technologies, such as polymerase chain reaction (PCR) and immunohistochemistry, have produced conflicting results for MCPyV infections in non-MCC tumors. Therefore, we performed quantitative analyses of the MCPyV copy number in various skin tumor tissues, including MCC (n?=?9) and other sun exposure-related skin tumors (basal cell carcinoma BCC, n?=?45], actinic keratosis AK, n?=?52], Bowen's disease n?=?34], seborrheic keratosis n?=?5], primary cutaneous anaplastic large-cell lymphoma n?=?5], malignant melanoma n?=?5], and melanocytic nevus n?=?6]). In a conventional PCR analysis, MCPyV DNA was detected in MCC (9 cases; 100%), BCC (1 case; 2%), and AK (3 cases; 6%). We then used digital PCR technology to estimate the absolute viral copy number per haploid human genome in these tissues. The viral copy number per haploid genome was estimated to be around 1 in most MCC tissues, and there were marked differences between the MCC (0.119-42.8) and AK (0.02-0.07) groups. PCR-positive BCC tissue showed a similar viral load as MCC tissue (0.662). Immunohistochemistry with a monoclonal antibody against the MCPyV T antigen (CM2B4) demonstrated positive nuclear localization in most of the high-viral-load tumor groups (8 of 9 MCC and 1 BCC), but not in the low-viral-load or PCR-negative tumor groups. These results demonstrated that MCPyV infection is possibly involved in a minority of sun-exposed skin tumors, including BCC and AK, and that these tumors display different modes of infection.
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