Generation of an infectious cDNA of a highly cardiovirulent coxsackievirus B3(CVB3m) and comparison to other infectious CVB3 cDNAs

Lee, C; Maull, Elizabeth A.; Chapman, Nora M.; Tracy, Steven; Wood, Jeffery T.; Gauntt, Charles J.

doi:10.1016/s0168-1702(97)00059-2

Cited by 25 publications

(22 citation statements)

References 43 publications

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“…There are currently five full-length infectious CVB3 cDNA clones for which complete genomic sequence data are available. These include four cardiovirulent strains (19,22,23,46) and one artificially attenuated noncardiovirulent strain (5). Nucleotide sequence analysis indicates that all five are highly related (37).…”

Section: Resultsmentioning

confidence: 99%

“…The approach taken in this paper is the only one possible at present, for there exists no amyocarditic strain of CVB3 that has been cloned as an infectious cDNA copy. All infectious cDNA clones of CVB3 genomes are from strains that either are naturally cardiovirulent (19,22,23,46) or, in the case of CVB3/0, can be made to be cardiovirulent if engineered to revert the C3U 234 mutation (5,6). In general, the nucleotide sequences of recent clinical CVB3 isolates differ significantly from the published full-length sequences in both the 5Ј NTR (37) and capsid coding region (Romero, unpublished data).…”

Section: Vol 74 2000 Determinants Of Cardiovirulence In Cvb3 Isolatmentioning

confidence: 99%

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Genomic Determinants of Cardiovirulence in Coxsackievirus B3 Clinical Isolates: Localization to the 5′ Nontranslated Region

Dunn

Chapman

Tracy

et al. 2000

J Virol

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Coxsackievirus B3 (CVB3) infections can cause myocarditis in humans and are implicated in the pathogenesis of dilated cardiomyopathy. The natural genetic determinants of cardiovirulence for CVB3 have not been identified, although using strains engineered in the laboratory, cardiovirulence determinants have been identified in the CVB3 5 nontranslated region (5NTR) and capsid. The myocarditic phenotypes of two CVB3 clinical isolates were determined using an established murine model of inflammatory heart disease. The 5NTRs and capsid proteins of the noncardiovirulent CVB3/CO strain and cardiovirulent CVB3/AS strain were examined to determine their influence on the cardiovirulence phenotype. Six intratypic chimeric viruses were constructed in which 5NTR and capsid sequences of the infectious cDNA copy of the cardiovirulent CVB3/20 genome were replaced by homologous sequences from CVB3/CO or CVB3/AS. Chimeric strains were tested for cardiovirulence by inoculation of C3H/HeJ mice. Sections of hearts removed at 10 days postinoculation were examined for evidence of myocarditis by light microscopy and assayed for the presence of virus. Replacement of the CVB3/ 20 capsid coding region by that from the homologous region of CVB3/CO resulted in no change in the cardiovirulent CVB3/20 phenotype, with virus recoverable from the heart at 10 days postinoculation. However, recombinant virus containing the CVB3/CO 5NTR alone or the 5NTR and capsid sequences together were not myocarditic, and infectious virus was not recovered from the myocardium. Chimeric viruses containing the CVB3/ AS 5NTR alone, capsid sequence alone, or both together preserved the myocarditic phenotype. These data support the 5NTR as the primary site in the determination of the natural cardiovirulence phenotype of CVB3.Coxsackievirus B3 (CVB3), one of six CVB serotypes, is a member of the genus Enterovirus within the family Picornaviridae. The genome of CVB3, like that of other enteroviruses, is a single-stranded, messenger sense, polyadenylated RNA molecule 7,400 nucleotides (nt) in length. The single open reading frame, flanked 5Ј and 3Ј by nontranslated regions (NTRs), is subdivided into three regions, P1 to P3 (51), that encode a polyprotein of 2,185 amino acids (aa). The P1 region encodes the viral capsid proteins VP1 (281 aa), VP2 (263 aa), VP3 (238 aa), and VP4 (69 aa). The P2 and P3 regions encode the nonstructural viral proteins important for processing of the polyprotein, replication, and translation. The near atomic structure of the CVB3 virion has been solved (31), demonstrating a structure similar to that of poliovirus type 1 (18) and rhinovirus (38).Coxsackie B viruses have been identified as a major cause of human viral myocarditis. They are also thought to play a significant role in the development of dilated cardiomyopathy (reviewed in reference 1). In general, neonates and children suffer more severe clinical syndromes due to CVB. In one study, of 41 infants who succumbed to CVB infection, approximately 95% demonstrated clinical or histologic...

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Section: Resultsmentioning

confidence: 99%

Section: Vol 74 2000 Determinants Of Cardiovirulence In Cvb3 Isolatmentioning

confidence: 99%

Genomic Determinants of Cardiovirulence in Coxsackievirus B3 Clinical Isolates: Localization to the 5′ Nontranslated Region

Dunn

Chapman

Tracy

et al. 2000

J Virol

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“…CVB4, strain JVB/Benschoten, was purchased from the American Type Culture Collection (Manassas, Va.) as a virus stock. The RNA genomes of coxsackievirus type B3, strains M and 20, have been cloned as infectious cDNA copies (67,103). The genome of CVB3/28 (pCVB3-28 [106]) was derived from the infectious cDNA clone of CVB3 strain 0 (CVB3/0 [19]) by alteration of nucleotide 234 from C to T; the resultant virulence phenotype is that of the CVB3/20 virus strain.…”

Section: Methodsmentioning

confidence: 99%

Toward Testing the Hypothesis that Group B Coxsackieviruses (CVB) Trigger Insulin-Dependent Diabetes: Inoculating Nonobese Diabetic Mice with CVB Markedly Lowers Diabetes Incidence

Tracy

Drescher

Chapman

et al. 2002

J Virol

147

164

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Insulin-dependent (type 1) diabetes mellitus (T1D) onset is mediated by individual human genetics as well as undefined environmental influences such as viral infections. The group B coxsackieviruses (CVB) are commonly named as putative T1D-inducing agents. We studied CVB replication in nonobese diabetic (NOD) mice to assess how infection by diverse CVB strains affected T1D incidence in a model of human T1D. Inoculation of 4-or 8-week-old NOD mice with any of nine different CVB strains significantly reduced the incidence of T1D by 2-to 10-fold over a 10-month period relative to T1D incidences in mock-infected control mice. Greater protection was conferred by more-pathogenic CVB strains relative to less-virulent or avirulent strains. Two CVB3 strains were employed to further explore the relationship of CVB virulence phenotypes to T1D onset and incidence: a pathogenic strain (CVB3/M) and a nonvirulent strain (CVB3/GA). CVB3/M replicated to four-to fivefold-higher titers than CVB3/GA in the pancreas and induced widespread pancreatitis, whereas CVB3/GA induced no pancreatitis. Apoptotic nuclei were detected by TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) assay in CVB3/M-infected pancreata but not in CVB3/GA-infected pancreata. In situ hybridization detected CVB3 RNA in acinar tissue but not in pancreatic islets. Although islets demonstrated inflammatory infiltrates in CVB3-protected mice, insulin remained detectable by immunohistochemistry in these islets but not in those from diabetic mice. Enzyme-linked immunosorbent assay-based examination of murine sera for immunoglobulin G1 (IgG1) and IgG2a immunoreactivity against diabetic autoantigens insulin and HSP60 revealed no statistically significant relationship between CVB3-protected mice or diabetic mice and specific autoimmunity. However, when pooled sera from CVB3/M-protected mice were used to probe a Western blot of pancreatic proteins, numerous proteins were detected, whereas only one band was detected by sera from CVB3/GA-protected mice. No proteins were detected by sera from diabetic or normal mice. Cumulatively, these data do not support the hypothesis that CVB are causative agents of T1D. To the contrary, CVB infections provide significant protection from T1D onset in NOD mice. Possible mechanisms by which this virus-induced protection may occur are discussed.The group B coxsackieviruses (CVB; family Picornaviridae, genus Enterovirus, species group B coxsackievirus; six serotypes, CVB1 to -6) are among the best studied of human enteroviruses (102). The CVB genome is a single strand of positive sense RNA 7,400 nucleotides in length that encodes 11 proteins in a single open reading frame (89). The CVB have been associated with diverse human diseases, among the more serious of which are myocarditis, pancreatitis, and aseptic meningitis. The CVB have been soundly implicated as causes of human myocarditis (1, 26, 42, 60-62, 73, 74, 108, 109) and pancreatitis (2,41,54,58,66,107) and, furthermore, cause these diseases readily i...

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“…Cardiovirulent CVB3 strains, including CVB3 20 , induced severe inflammatory lesions, and virus replication persisted through day 14 after injection in the heart [15,16] . Other tissues, such as those of the liver and pancreas, were also damaged after the recovery of infectious virus.…”

Section: Introductionmentioning

confidence: 99%

Mutation Variants Generated from Nonvirulent Coxsackievirus B3 Acquire Virulence Phenotypes by Active Virus Replication

Seo¹,

Jee

Ahn³

et al. 2007

Intervirology

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Objective: To understand coxsackievirus B3 (CVB3) virulence at the molecular level. Method: A mutation library was generated from noncardiovirulent CVB3/0. Highly virulent mutation variants were recovered and characterized both phenotypically and genotypically. Results: The variants consistently caused destruction of multiple tissues together with active virus production and induced severe mortality in vivo. The extent of infectious virus generation was directly correlated with that of histopathological aberration. Genotypic analysis of the entire genome indicated that the virulent viruses encode nucleotide substitutions in the 5′-nontranslated region, which have previously been identified in other virulent CVB3s. Conclusion: The present study provides evidence that particular nucleotide substitutions in the 5′-nontranslated region of nonvirulent CVB3 can lead to active virus replication, which is sufficient to induce virulence.

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Generation of an infectious cDNA of a highly cardiovirulent coxsackievirus B3(CVB3m) and comparison to other infectious CVB3 cDNAs

Cited by 25 publications

References 43 publications

Genomic Determinants of Cardiovirulence in Coxsackievirus B3 Clinical Isolates: Localization to the 5′ Nontranslated Region

Genomic Determinants of Cardiovirulence in Coxsackievirus B3 Clinical Isolates: Localization to the 5′ Nontranslated Region

Toward Testing the Hypothesis that Group B Coxsackieviruses (CVB) Trigger Insulin-Dependent Diabetes: Inoculating Nonobese Diabetic Mice with CVB Markedly Lowers Diabetes Incidence

Mutation Variants Generated from Nonvirulent Coxsackievirus B3 Acquire Virulence Phenotypes by Active Virus Replication

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