Isolation and Type Determination of Coxsackie Virus, Group B, in Tissue Culture

Sickles, Grace M.; Feorino, Paul M.; Plager, Hildegarde

doi:10.3181/00379727-88-21481

Cited by 18 publications

(6 citation statements)

References 1 publication

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“…Mice were intraperitoneally injected with normal saline solution (control) or 1 × 10 4 plaque-forming units (pfu) of the prototypical CVB4 laboratory strain JVB (catalog # VR-184; American Type Culture Collection) grown in HeLa cells ( 20 ). Nonfasting blood glucose levels were measured at least twice weekly.…”

Section: Methodsmentioning

confidence: 99%

“…The CodeSet included type I IFN, cytokines, apoptosis, endocrine, endoplasmic reticulum (ER) stress, T1D-associated loci, and other human genes, plus seven housekeeping genes for normalization of data. A probe for a conserved CVB sequence targeting the same region as the quantitative RT-PCR primer ( 20 ) was included. One hundred nanograms of RNA extracted from tissue was hybridized, processed, and analyzed per the manufacturer’s procedure.…”

Section: Methodsmentioning

confidence: 99%

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Viral Infection of Engrafted Human Islets Leads to Diabetes

et al. 2014

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Type 1 diabetes (T1D) is characterized by the destruction of the insulin-producing β-cells of pancreatic islets. Genetic and environmental factors both contribute to T1D development. Viral infection with enteroviruses is a suspected trigger for T1D, but a causal role remains unproven and controversial. Studies in animals are problematic because of species-specific differences in host cell susceptibility and immune responses to candidate viral pathogens such as coxsackievirus B (CVB). In order to resolve the controversial role of viruses in human T1D, we developed a viral infection model in immunodeficient mice bearing human islet grafts. Hyperglycemia was induced in mice by specific ablation of native β-cells. Human islets, which are naturally susceptible to CVB infection, were transplanted to restore normoglycemia. Transplanted mice were infected with CVB4 and monitored for hyperglycemia. Forty-seven percent of CVB4-infected mice developed hyperglycemia. Human islet grafts from infected mice contained viral RNA, expressed viral protein, and had reduced insulin levels compared with grafts from uninfected mice. Human-specific gene expression profiles in grafts from infected mice revealed the induction of multiple interferon-stimulated genes. Thus, human islets can become severely dysfunctional with diminished insulin production after CVB infection of β-cells, resulting in diabetes.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Viral Infection of Engrafted Human Islets Leads to Diabetes

et al. 2014

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“…The relatively low identity with the prototype CVB2 sequence (and to other prototype CVB genomes) in comparison to the 2002 Kanagawa CVB2 isolates, was not surprising given that the prototype strains circulated more than 50 years ago (Melnick et al, 1950). However, a recently published study of an HEV in type I diabetes patients (Dotta et al, 2007) generated a sequence which shared >99% overall nt identity with the CVB4 prototype strain Benschoten originally isolated in 1951 (Sickles, Feorino, and Plager, 1955), with just one nt mismatch in the 5'NTR. As RT-PCR has a risk of contamination with laboratory strains [see, for example, (Giacca et al, 1994)], it can be concluded that the CVB4, "Tuscany" strain (Dotta et al, 2007), was derived from contamination with the 58 year old prototype strain.…”

Section: The Nucleotide and Amino Acid Sequences Of The Heart Virus Imentioning

confidence: 97%

5′ terminal deletions in the genome of a coxsackievirus B2 strain occurred naturally in human heart

et al. 2008

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Enteroviruses can induce human myocarditis, which can be modeled in mice inoculated with group B coxsackieviruses (CVB) and in which CVB evolve to produce defective, terminally deleted genomes. The 5' non-translated region (NTR) was enzymatically amplified from heart tissue of a fatal case of enterovirus-associated myocarditis in Japan in 2002. While no intact 5' viral genomic termini were detected, 5' terminal deletions ranged in size from 22 to 36 nucleotides. Sequence of the 5' third of this viral genome is of a modern strain, closely related to CVB2 strains isolated in Japan in 2002. A CVB3 chimera containing the 5' NTR with a 22 nt deletion produced progeny virus upon transfection of HeLa cells. When the 5' 22 nucleotide deletion was repaired, the virus induced myocarditis in mice and replicated like wild type virus in murine heart cells. This is the first report of these naturally-occurring defective enteroviral genomes in human myocarditis.

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“…(8)(9)(10), attempts were made to titrate adult mouse 1302 virus by this convenient method. Surprisingly, cytopathogenic effects were not noted by the seventh day in either human (Hela) or Monkey (kidney) cell cultures, despite the demonstrable cytopathogenicity of the infant mouse line of the same virus (Table V).…”

Section: Myocardial Lesions In Control Mice Injected With Cortisonementioning

confidence: 99%