title: Mechanisms of beta cell death during restricted and unrestricted enterovirus infection.
abstract: Coxsackie B virus (CVB-5) infections potentially trigger and accelerate pancreatic beta cell damage leading to type 1 diabetes. In vivo, all viruses face natural resistance mediated by various host factors which restrict the progression of infection. Thus, the aims of this study were to generate a tissue culture model of restricted coxsackie B virus infection in primary islet cells by preventing the production of viral progeny with a selective inhibitor of viral RNA replication and to investigate the mechanisms of virus-induced islet cell death during productive and restricted infective conditions. Cultured foetal porcine islet cells were infected effectively with the prototype strain of coxsackievirus B5 (CVB-5). Nuclear viability stainings and electron microscopy showed productive infection to result in dominantly necrotic cell death with additional slight induction of apoptosis during the 7 days of follow-up. The restricted conditions were created by addition of guanidine-hydrochloride (G-HCl) into culture medium. At 1 mM concentration, it significantly protected the infected cells from necrosis and thus maintained high viability. This was associated with increased significantly apoptosis. In perifusion analysis, the cellular ability to release insulin was reduced, although the metabolic integrity was preserved as shown by MTT-analysis and cellular ATP levels. These data show that restriction of CVB-5 replication with G-HCl protects islet cells against virus-induced necrosis. However, restriction of viral replication shifts the mechanism of cell death from necrosis toward apoptosis. A slowly progressing subclinical infection of islets could thus lead to increased beta-cell apoptosis.
authors.affiliation: Haartman Institute, Transplantation Laboratory, University of Helsinki, Helsinki, Finland. suvi.rasilainen@helsinki.fi
authors.forename: Suvi
authors.initials: S
authors.lastname: Rasilainen
authors.forename: Petri
authors.initials: P
authors.lastname: Ylipaasto
authors.forename: Merja
authors.initials: M
authors.lastname: Roivainen
authors.forename: Luc
authors.initials: L
authors.lastname: Bouwens
authors.forename: Risto
authors.initials: R
authors.lastname: Lapatto
authors.forename: Tapani
authors.initials: T
authors.lastname: Hovi
authors.forename: Timo
authors.initials: T
authors.lastname: Otonkoski
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017209:Apoptosis; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D046508:Culture Techniques; D029822:Enterovirus B, Human / Q000254:growth & development / Q000472:pathogenicity*; D004769:Enterovirus Infections / Q000473:pathology* / Q000821:virology; D019791:Guanidine / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure / Q000821:virology*; D009336:Necrosis; D013552:Swine; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects
chemical_list: D007328:Insulin; D013778:Tetrazolium Salts; D013844:Thiazoles; D000255:Adenosine Triphosphate; C022616:thiazolyl blue; D019791:Guanidine