Ongoing enterovirus-induced myocarditis is associated with persistent heart muscle infection: quantitative analysis of virus replication, tissue damage, and inflammation.

Abstract

Coxsackievirus B3-induced myocarditis in different immunocompetent mouse strains was used as a model to investigate interrelationships between virus replication and development of chronic enteroviral heart disease. Using in situ hybridization to detect enteroviral RNA, we show that heart muscle infection is not only detected in acute myocarditis but is also detected during the chronic phase of the disease. Coxsackievirus B3 could evade immunological surveillance in a host-dependent fashion, thus inducing a persistent infection of the myocardium in association with ongoing inflammation. Patterns of acute and persistent myocardial infection were quantitatively assessed in one representative mouse strain (A.CA/SnJ, H-2f) by applying computer-assisted digital image processing; these patterns were then related to the extent of myocardial tissue damage as well as to inflammation. We observed a strong correlation, both spatial and temporal, between viral replication and development of myocardial lesions, indicating that acute and chronic myocardial injuries are a consequence of multifocal organ infection. Analysis of strandspecific in situ hybridization revealed that viral replication in persistent infection is restricted at the level of RNA synthesis. The described procedure for quantitating organ infection provides a powerful tool for evaluating virus-host interactions and will be of particular interest to those studying human enterovirus-induced cardiomyopathies.

The prevalence of enteroviral capsid protein vp1 immunostaining in pancreatic islets in human type 1 diabetes

“…By contrast, when the infection continued into a chronic phase, the number of positive and negative enteroviral RNA strands per cell became equivalent and capsid protein was less readily demonstrable [32]. On this basis, it is conceivable that a molecular form of virus that is not readily detectable by immunohistochemistry for vp1, may exist as a chronic infection in many of the beta cells in type 1 diabetes and that, occasionally, production of the whole virus increases, leading to capsid protein synthesis in a small number of beta cells.…”

Antigen‐presentation capacity of dendritic cells is impaired in ongoing enterovirus myocarditis

“…Correspondence: Dr. Peter M. Kloetzel e-mail: p-m.kloetzel@charite.de ongoing disease with myocardial scarring and inflammatory lesions in areas of virus persistence [3].…”

“…Ongoing research focuses on the delineation of host determinants that identify patients at risk for the development of chronic disease. The murine model of ongoing CVB3-myocarditis properly mimics human disease progression [3][4][5][6][7]: upon CVB3 infection so-called resistant C57BL/6 mice eliminate the virus following acute myocarditis. In contrast, MHC-matched, immunocompetent A.BY/SnJ mice develop ongoing chronic myocarditis with chronic virus persistence [3,6].…”

Expression of Ksp-cadherin during kidney development and in renal cell carcinoma

“…The obtained linearised DNA was purified by chloroform/phenol extraction and served subsequently as a template for preparation of single-stranded 35 Slabelled RNA probes, which were synthesised using SP6 RNA polymerase. Enterovirus-specific single-stranded RNA probes were used as negative controls (Klingel et al, 1992).…”

“…In brief, slides were pretreated with 0.2 M HCl, heated and digested with proteinase K. Then, the slides were dehydrated in graded ethanol series, dried over night and hybridised at 421C for 18 h with 500 ng ml À1 35 S-labelled RNA. After washing, nonhybridised single-stranded RNA was digested with RNase A as previously described (Klingel et al, 1992). Hybridised preparations were autoradiographed with Kodak NBT2 film, exposed for several weeks, and stained with haematoxylin/ eosin.…”