Phenotypic and genetic changes in coxsackievirus B5 following repeated passage in mouse pancreas in vivo

Abstract

Common enterovirus infections appear to initiate or facilitate the pathogenetic processes leading to type 1 diabetes, and also sometimes precipitate the clinical disease. In experimental infection of mice, coxsackieviruses have shown to have a strong affinity for the exocrine tissue, while even in lethal cases, the islets remain unaffected. The virus strain most intensively studied in this respect is the diabetogenic variant E2 of coxsackievirus B4. In addition, it is known that all six serotypes of coxsackie B viruses can be made diabetogenic by repeated passages in either mouse pancreas in vivo or in cultured mouse beta-cells in vitro. However, the genetic determinants of the phenomenon have not been determined. In the present study, a laboratory strain of coxsackievirus B5 was passaged repeatedly in mouse pancreas in vivo. After 15 passages, the virus phenotype was clearly changed and infection of the variant resulted in a diabetes-like syndrome in mice characterized by chronic pancreatic inflammation together with dysregulation in glucose metabolism, loss of pancreatic acinar tissue, and mild insulitis. In order to characterize the genetic determinants involved in mouse pancreas adaptation, the passaged virus variant together with the parental virus strain was cloned for molecular characterization. The whole genome sequencing of both virus strains revealed only limited differences. Altogether, eight nucleotides were changed resulting in five amino acid substitutions, of which three were located in the capsid proteins.

Enterovirus RNA sequences in sera of schoolchildren in the general population and their association with type 1-diabetes-associated autoantibodies

“…Molecular characterization revealed limited genetic changes located not only in the capsid protein involved in virus-cell interaction but also in the non-structural proteins. This would mean that mutations in the non-structural genes may also affect virulence (Al-Hello et al, 2005).In conclusion these results based on a normal schoolchild population without T1D first-degree relatives support the role of EV infections in the initiation of T1D and the necessity of establishing concepts for prevention and/or treatment of EV infections. …”

“…Molecular characterization revealed limited genetic changes located not only in the capsid protein involved in virus-cell interaction but also in the non-structural proteins. This would mean that mutations in the non-structural genes may also affect virulence (Al-Hello et al, 2005).…”

Enterovirus-induced gene expression profile is critical for human pancreatic islet destruction

“…Viruses CVB-5-DS [24] and molecular clones derived from echovirus 9 strain DM (E-9-DM) [9,17] were used in the study. The infective clones were constructed as described previously [25].…”

Enterovirus strain and type-specific differences in growth kinetics and virus-induced cell destruction in human pancreatic duct epithelial HPDE cells

“…Yoon. The production of CV-B5-DS strain, mouse pancreas (in vivo) adapted CV-B5-MPA strain and murine insulinoma cell adapted CV-B5-MCA strain is described elsewhere (Al-Hello et al, 2005). The strains E-9-DM (Vreugdenhil et al, 2000) and E-11-D207 (Al-Hello et al, 2008) were originally isolated from patients at the clinical onset of type 1 diabetes and passaged twice in GMK cells.…”