Characterization of an infectious cDNA copy of the genome of a naturally occurring, avirulent coxsackievirus B3 clinical isolate

• Published in: Journal of general virology Vol. 86; no. 1; pp. 197 - 210
• Main Authors: Lee, C.-K, Kono, K, Haas, E, Kim, K.-S, Drescher, K. M, Chapman, N. M, Tracy, S
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01-01-2005; Society for General Microbiology
• Subjects: 5' Untranslated Regions - genetics; Animals; Base Sequence; Biological and medical sciences; Coxsackievirus B3; Disease Models, Animal; DNA, Complementary - chemistry; DNA, Complementary - genetics; Enterovirus B, Human - genetics; Enterovirus B, Human - immunology; Enterovirus B, Human - pathogenicity; Enterovirus Infections - prevention & control; Enterovirus Infections - virology; Fundamental and applied biological sciences. Psychology; Genome, Viral; Humans; Male; Mice; Mice, Inbred C3H; Microbiology; Miscellaneous; Molecular Sequence Data; Myocarditis - prevention & control; Myocarditis - virology; Pancreatitis - prevention & control; Pancreatitis - virology; Phenotype; RNA, Viral - genetics; Sequence Alignment; Virology; Virulence - genetics; Virus; Characterization; Infection; Complementary DNA; Microbiology; Picornaviridae; Coxsackievirus B3; Coxsackievirus A; Enterovirus; Genome; Clinical isolate; Virology
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/vir.0.80424-0

1 Enterovirus Research Laboratory, Department of Pathology and Microbiology, University of Nebraska Medical Center, 986495 Nebraska Medical Center, Omaha, NE 68198, USA
2 Department of Pathology and Microbiology, University of Nebraska Medical Center, 986495 Nebraska Medical Center, Omaha, NE 68198, USA
3 Department of Medical Microbiology and Immunology, Creighton University School of Medicine, Omaha, NE 68178, USA
Correspondence S. Tracy stracy{at}unmc.edu
Group B coxsackieviruses (CVB) cause numerous diseases, including myocarditis, pancreatitis, aseptic meningitis and possibly type 1 diabetes. To date, infectious cDNA copies of CVB type 3 (CVB3) genomes have all been derived from pathogenic virus strains. An infectious cDNA copy of the well-characterized, non-pathogenic CVB3 strain GA genome was cloned in order to facilitate mapping of the CVB genes that influence expression of a virulence phenotype. Comparison of the sequence of the parental CVB3/GA population, derived by direct RT-PCR-mediated sequence analysis, to that of the infectious CVB3/GA progeny genome demonstrated that an authentic copy was cloned; numerous differences were observed in coding and non-coding sequences relative to other CVB3 strains. Progeny CVB3/GA replicated similarly to the parental strain in three different cell cultures and was avirulent when inoculated into mice, causing neither pancreatitis nor myocarditis. Inoculation of mice with CVB3/GA protected mice completely against myocarditis and pancreatitis induced by cardiovirulent CVB3 challenge. The secondary structure predicted for the CVB3/GA domain II, a region within the 5' non-translated region that is implicated as a key site affecting the expression of a cardiovirulent phenotype, differs from those predicted for cardiovirulent and pancreovirulent CVB3 strains. This is the first report characterizing a cloned CVB3 genome from an avirulent strain.
These authors contributed equally to this work.
The GenBank/EMBL/DDBJ accession number for the sequence reported in this paper is AY673831 .
Present address: Department of Laboratory Medicine, Korea University Guro Hospital, Guro-gu Guro-dong 80, 152-050 Seoul, Republic of Korea.