Newly identified human rhinoviruses: molecular methods heat up the cold viruses

Arden, Katherine E.; Mackay, Ian M.

doi:10.1002/rmv.644

Cited by 79 publications

(75 citation statements)

References 161 publications

(278 reference statements)

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“…For example, bacterial surface polysaccharides stabilize polioviruses and facilitate cell attachment by increasing binding to the viral receptor (33,34). In any case, experiments in our laboratory and elsewhere demonstrated that the RV-C interaction with native HeLa cells or other cell lines (e.g., A549) is not sufficient for efficient virus entry and replication (5,6,35).…”

Section: Discussionmentioning

confidence: 89%

“…Viruses belonging to the RV-C species were only discovered in 2006 (1,2), some 50 y after the initial identification of other RVs (3), because these viruses are not detectable by standard tissue-culture techniques (4)(5)(6)(7). RV-Cs are of special clinical interest because they can cause more severe illnesses requiring hospitalization in infants and children compared with the RV-A or RV-B, and are closely associated with acute exacerbations of asthma (8)(9)(10).…”

mentioning

confidence: 99%

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Cadherin-related family member 3, a childhood asthma susceptibility gene product, mediates rhinovirus C binding and replication

Bochkov¹,

Watters

Ashraf³

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

371

345

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Members of rhinovirus C (RV-C) species are more likely to cause wheezing illnesses and asthma exacerbations compared with other rhinoviruses. The cellular receptor for these viruses was heretofore unknown. We report here that expression of human cadherin-related family member 3 (CDHR3) enables the cells normally unsusceptible to RV-C infection to support both virus binding and replication. A coding single nucleotide polymorphism (rs6967330, C529Y) was previously linked to greater cell-surface expression of CDHR3 protein, and an increased risk of wheezing illnesses and hospitalizations for childhood asthma. Compared with wild-type CDHR3, cells transfected with the CDHR3-Y529 variant had about 10-fold increases in RV-C binding and progeny yields. We developed a transduced HeLa cell line (HeLa-E8) stably expressing CDHR3-Y529 that supports RV-C propagation in vitro. Modeling of CDHR3 structure identified potential binding sites that could impact the virus surface in regions that are highly conserved among all RV-C types. Our findings identify that the asthma susceptibility gene product CDHR3 mediates RV-C entry into host cells, and suggest that rs6967330 mutation could be a risk factor for RV-C wheezing illnesses.

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Section: Discussionmentioning

confidence: 89%

mentioning

confidence: 99%

Cadherin-related family member 3, a childhood asthma susceptibility gene product, mediates rhinovirus C binding and replication

Bochkov¹,

Watters

Ashraf³

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

371

345

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“…Clinically and biologically, they share many attributes with the other designated HRV species, HRV-A and -B. Most studies of HRV-C disease associations, typically focused on children from asthmatic and/or hospital-based populations (Arden & Mackay, 2010), have demonstrated a similarly broad range of clinical outcomes to those observed in HRV-A and -B infections and, indeed, with other respiratory viruses. Some studies show no difference in clinical outcome between HRV species (Lau et al, 2007;Piotrowska et al, 2009), whereas others provide evidence for a more frequent role of HRV-C in lower respiratory tract disease, febrile wheeze in infants and toddlers, and asthma exacerbations in older children (Lau et al, 2007;Khetsuriani et al, 2008;Miller et al, 2009a;Wisdom et al, 2009b).…”

Section: Introductionmentioning

confidence: 99%

“…Clinically and biologically, they share many attributes with the other designated HRV species, HRV-A and -B. Most studies of HRV-C disease associations, typically focused on children from asthmatic and/or hospital-based populations (Arden & Mackay, 2010), have demonstrated a similarly broad range of clinical outcomes to those observed in HRV-A and -B infections and, indeed, with other respiratory viruses. Some A full list of all HRV-C variants characterized to date, categorized into a total of 61 confirmed or provisionally assigned types, is available with the online version of this paper.…”

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confidence: 99%

Proposals for the classification of human rhinovirus species C into genotypically assigned types

Simmonds

McIntyre

Savolainen-Kopra

et al. 2010

Journal of General Virology

212

163

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Human rhinoviruses (HRVs) are common respiratory pathogens associated with mild upper respiratory tract infections, but also increasingly recognized in the aetiology of severe lower respiratory tract disease. Wider use of molecular diagnostics has led to a recent reappraisal of HRV genetic diversity, including the discovery of HRV species C (HRV-C), which is refractory to traditional virus isolation procedures. Although it is heterogeneous genetically, there has to date been no attempt to classify HRV-C into types analogous to the multiple serotypes identified for HRV-A and -B and among human enteroviruses. Direct investigation of cross-neutralization properties of HRV-C is precluded by the lack of methods for in vitro culture, but sequences from the capsid genes (VP1 and partial VP4/VP2) show evidence for marked phylogenetic clustering, suggesting the possibility of a genetically based system comparable to that used for the assignment of new enterovirus types. We propose a threshold of 13 % divergence for VP1 nucleotide sequences for type assignment, a level that classifies the current dataset of 86 HRV-C VP1 sequences into a total of 33 types. We recognize, however, that most HRV-C sequence data have been collected in the VP4/VP2 region (currently 701 sequences between positions 615 and 1043). We propose a subsidiary classification of variants showing .10 % divergence in VP4/ VP2, but lacking VP1 sequences, to 28 provisionally assigned types (subject to confirmation once VP1 sequences are determined). These proposals will assist in future epidemiological and clinical studies of HRV-C conducted by different groups worldwide, and provide the foundation for future exploration of type-associated differences in clinical presentations and biological properties. IntroductionHuman rhinoviruses (HRVs) are highly prevalent respiratory pathogens, most commonly associated with mild upper respiratory tract disease and exacerbations of preexisting respiratory disease such as asthma. They are also increasingly recognized as underlying more severe disease manifestations, such as bronchiolitis in young children and in the immunosuppressed. The increasing use of molecular methods for respiratory virus screening has contributed to this reappraisal of rhinoviruses, as has the recent discovery of an entirely novel rhinovirus group, refractory to previously used virus isolation methods but now known to be highly prevalent and widely circulating worldwide (Arden et al., 2006;Kaiser et al., 2006;Lamson et al., 2006;Kistler et al., 2007;Lau et al., 2007;Lee et al., 2007;McErlean et al., 2007;Renwick et al., 2007;Olenec et al., 2010).These newly characterized rhinoviruses have been proposed to belong to a novel species of rhinovirus (designated species C; HRV-C), recognizing their substantial sequence divergence from other classified species within the genus Enterovirus of picornaviruses (Carstens, 2010;Knowles, 2010) (Fig. 1a). Clinically and biologically, they share many attributes with the other designated HRV species, HRV-A and -B....

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“…One disadvantage associated with the use of RV 5=-UTR primers is nonspecific amplification of human genomic DNA (chromosome 6) or RNA (large regulator noncoding RNA B2) sequences, yielding a nonspecific 424-bp product that is similar in size to the virus-specific amplicon (390 bp) (19)(20)(21). The nonspecific results are typically found in a small subset of nasal lavage fluid samples but are more common when clinical samples contain high concentrations of human RNA or contaminating genomic DNA (e.g., cases of intense cellular inflammation in airways or nasal brushing samples).…”

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confidence: 99%

Improved Molecular Typing Assay for Rhinovirus Species A, B, and C

et al. 2014

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Human rhinoviruses (RVs), comprising three species (A, B, and C) of the genus Enterovirus, are responsible for the majority of upper respiratory tract infections and are associated with severe lower respiratory tract illnesses such as pneumonia and asthma exacerbations. High genetic diversity and continuous identification of new types necessitate regular updating of the diagnostic assays for the accurate and comprehensive detection of circulating RVs. Methods for molecular typing based on phylogenetic comparisons of a variable fragment in the 5= untranslated region were improved to increase assay sensitivity and to eliminate nonspecific amplification of human sequences, which are observed occasionally in clinical samples. A modified set of primers based on new sequence information and improved buffers and enzymes for seminested PCR assays provided higher specificity and sensitivity for virus detection. In addition, new diagnostic primers were designed for unequivocal species and type assignments for RV-C isolates, based on phylogenetic analysis of partial VP4/VP2 coding sequences. The improved assay was evaluated by typing RVs in >3,800 clinical samples. RVs were successfully detected and typed in 99% of the samples that were RV positive in multiplex diagnostic assays. Rhinoviruses (RVs) are members of the genus Enterovirus of the family Picornaviridae and are currently classified into three species (A, B, and C). More than 160 RV types have been identified to date, including 99 classic serotypes of RV-A and RV-B (1) and 6, 5, and 51 novel genotypes of RV-A, RV-B, and RV-C, respectively, classified genetically in the absence of serological cross-neutralization data (2-4). RVs are responsible for the majority of upper respiratory tract infections (common colds) and also can cause more severe illnesses of the lower respiratory tract, such as pneumonia and asthma exacerbations. The species affects the virulence, as RV-A and RV-C cause more severe illnesses in infants and are more likely to cause exacerbations of childhood asthma (5, 6).RV is a positive-sense single-stranded RNA virus with a genome of 7.1 to 7.2 kb, consisting of a single gene that codes for a long polyprotein of about 2,100 amino acids. The translated polyprotein is cleaved by viral proteases 2A and 3C to yield 11 mature proteins. The viral capsid is composed of 60 copies each of the VP4, VP2, VP3, and VP1 proteins. There are two untranslated regions (UTRs), i.e., the 5= UTR, which is typically 610 to 630 bases long and precedes the open reading frame, and the 3= UTR, which consists of 40 to 45 bases upstream of the poly(A) tract. The 5= and 3= UTRs contain a number of structural and sequence elements necessary for viral genome translation and replication. Sequence analyses of RVs reveal high genetic diversity, especially in the capsid-coding region, and evidence for recombination events mapped mainly in the 5= UTR and the protease 2A gene (7,8).Molecular methods for viral genome detection and sequencing directly in clinical samples are essential f...

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Newly identified human rhinoviruses: molecular methods heat up the cold viruses

Cited by 79 publications

References 161 publications

Cadherin-related family member 3, a childhood asthma susceptibility gene product, mediates rhinovirus C binding and replication

Cadherin-related family member 3, a childhood asthma susceptibility gene product, mediates rhinovirus C binding and replication

Proposals for the classification of human rhinovirus species C into genotypically assigned types

Improved Molecular Typing Assay for Rhinovirus Species A, B, and C

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