Merkel cell polyomavirus expression in merkel cell carcinomas and its absence in combined tumors and pulmonary neuroendocrine carcinomas

Abstract

Merkel cell carcinoma (MCC) is the eponym for primary cutaneous neuroendocrine carcinoma. Recently, a new polyoma virus has been identified that is clonally integrated in the genome of the majority of MCCs, with truncating mutations in the viral large T antigen gene. We examined the presence of Merkel cell polyomavirus (MCV) in a set of 17 frozen tumor samples by quantitative polymerase chain reaction; 15 of them (88%) were positive. Sections from corresponding archival material were analyzed by immunohistochemistry (IHC) with the novel monoclonal antibody CM2B4, generated against a predicted antigenic epitope on the MCV T antigen, and tested for the expression of cytokeratin 20 (CK20). Sufficient archival material for IHC was available in only 15 of the 17 cases whose frozen tissue samples had been studied by polymerase chain reaction. Of the 15 tumors analyzed immunohistochemically, 10 (67%) showed positive labeling with CM2B4, 14 (93%) expressed CK20. A tissue microarray of 36 MCCs, 7 combined squamous and neuroendocrine carcinomas of the skin, and 26 pulmonary neuroendocrine carcinomas were also examined by IHC. Of the 36 MCCs assembled on a microarray, 32 (89%) tumors expressed CK20, and 27 (75%) were immunoreactive with CM2B4. The skin tumors with a combined squamous and neuroendocrine phenotype and all pulmonary neuroendocrine carcinomas failed to react with CM2B4. Our study shows that CM2B4 is a useful reagent for the diagnosis of MCC. It labels the majority of MCCs, but fails to react with pulmonary neuroendocrine carcinomas. We also found that neuroendocrine carcinomas of the skin arising in association with a squamous cell carcinoma seem to be independent of MCV.

FIGURE 1

A, Merkel cell carcinoma presenting as tumor nodule in the dermis and subcutis. B, Dense population of blue cells (hematoxylin and eosin-stained section). C, Positive staining for cytokeratin 20 in a predominantly paranuclear dot-like pattern. D, Nuclear labeling with monoclonal antibody CM2B4 (this tumor was polymerase chain reaction-positive for Merkel cell polyomavirus).

FIGURE 2

Tissue microarray—patterns of colabeling for CK20 and monoclonal antibody CM2B4. A and B, A tumor is positive for both CK20 (A) and CM2B4 (B) [most commonly seen]. C and D, A tumor negative for CK20 (C), but immunreactive for CM2B4 (D). E and F, A tumor expresses CK20 (E), but fails to react with CM2B4 (F). G and H, A tumor is negative for both CK20 (G) and CM2B4 (H). CK20 indicates cytokeratin 20.

FIGURE 3

Combined cutaneous squamous and neuroendocrine carcinoma. A, The tumor shows a biphenotypic appearance (hematoxylin and eosin-stained section). It contains a pink squamous and a small cell neuroendocrine component. B, The neuroendocrine component is immunoreactive for cytokeratin 20. C, The neuroendocrine component fails to react with CM2B4.