Localization of Enteroviral Antigen in Myocardium and Other Tissues from Patients with Heart Muscle Disease by an Improved Immunohistochemical TechniqueHongyi Zhanga,b, Yanwen Lia, Tianqing Penga,c, Malen Aasaa, Li Zhanga, Yingzhen Yangc, and Leonard C. Archardaa Molecular Pathology Section, Division of Biomedical Sciences, Imperial College of Science, Technology and Medicine, London, United Kingdom b Medical Microbiology, Division of Investigative Science, Imperial College of Science, Technology and Medicine, London, United Kingdom c Key Laboratory of Viral Heart Disease, Ministry of Public Health, Zhongshan Hospital, Shanghai Medical University, Shanghai, People's Republic of China Correspondence to: Hongyi Zhang, Molecular Pathology Section, Div. of Biomedical Sciences, SAF Building, ICSTM, South Kensington, London SW7 2AZ, UK. E-mail: h.zhang@ic.ac.uk The association of enterovirus infection and heart muscle diseases has been investigated extensively by detection of viral genomic RNA using nucleic acid hybridization and the reverse transcription-polymerase chain reaction. To further understand the role of enterovirus and its persistence in these diseases, an immunohistochemical technique was optimized to investigate the expression of viral capsid proteins in situ. A monoclonal antibody (5-D8/1) against an epitope in the N-terminus of capsid protein VP1, conserved in the enterovirus genus, was employed. To enhance sensitivity, the EnVison system was used to detect antigenantibody complex. VP1 was detected in formalin-fixed, paraffin-embedded endomyocardial biopsy or postmortem myocardial tissues and in liver, spleen, lung, kidney, and pancreas from patients with myocarditis or dilated cardiomyopathy, but not from controls. VP1 was localized in cytoplasm of myofibers, often adjacent to necrosis and infiltrate in myocarditis, and was clustered or scattered in dilated cardiomyopathy. This technique can be used for a definitive laboratory diagnosis of enterovirus-associated diseases and for studying the mechanisms of virus persistence in chronic myocardial disease. (J Histochem Cytochem 48:579584, 2000) Key Words: enterovirus, antigen, myocarditis, cardiomyopathy, immunohistochemistry, monoclonal antibody, EnVision
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