Self-Assembly of the Recombinant Capsid Protein of a Bovine Norovirus (BoNV) into Virus-Like Particles and Evaluation of Cross-Reactivity of BoNV with Human Noroviruses

Han, Myung Guk; Wang, Q.; Smiley, Jeffrey R.; Chang, Kyeong‐Ok; Saif, Linda J.

doi:10.1128/jcm.43.2.778-785.2005

Cited by 24 publications

(36 citation statements)

References 59 publications

(68 reference statements)

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“…The low value of Spearman's rank correlation indicated a moderate cross-reactivity of antibodies against NoV GIII to NoV GI and GII, but this alone could not explain the observed seroreactivity to bovine VLPs. This is similar to other studies which show limited cross-reactivity between human and bovine NoV strains (14,15,21,34). The differences in the prevalence rates of antibody to the VLPs tested likely indicate different levels of exposure to these viruses, with significant differences in exposure between human genogroups and bovine NoVs.…”

supporting

confidence: 87%

“…The GI and GII VLPs previously evaluated for specificity using the antisera prepared against the expressed VLPs showed that they were antigenically distinct (25). Studies have also shown the specificity of GIII and NB VLPs, with no cross-reactivity observed with GI and GII with the antiserum reagents produced (21,26). Sequence comparison has shown a low level of amino acid sequence identity between human and bovine norovirus strains, suggesting that antigenically and genetically, the strains are distinct (27).…”

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confidence: 98%

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Exposure to Human and Bovine Noroviruses in a Birth Cohort in Southern India from 2002 to 2006

et al. 2013

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cHuman and bovine norovirus virus-like particles were used to evaluate antibodies in Indian children at ages 6 and 36 months and their mothers. Antibodies to genogroup II viruses were acquired early and were more prevalent than antibodies to genogroup I. Low levels of IgG antibodies against bovine noroviruses indicate possible zoonotic transmission. N oroviruses (NoVs) are nonenveloped, single-stranded positive-sense RNA viruses, accounting for ϳ50% of gastroenteritis outbreaks worldwide (1). Seroepidemiological surveys using NoV virus-like particles (VLPs) as antigens show exposure to NoVs across the globe (2-7), with high seroprevalence in children Ͻ5 years of age; seroprevalence can reach 100% in adults (2,8,9). Analyses of bovine strains suggest close relation to human NoV genogroup I (GI) and . Studies from the Netherlands found that 22% of the population had antibodies to bovine NoV, with veterinarians having high frequencies of antibodies (15), raising the possibility of zoonotic transmission (16). In this study, sera from children in a birth cohort and their mothers were used to assess exposure to human and bovine norovirus genogroups in early life and adulthood.The study population was a birth cohort from semiurban slums in Vellore, South India, recruited and monitored from 2002 to 2006, with sample collection as previously described (17)(18)(19). Maternal sera at delivery and sera from children at 6 and 36 months were tested for IgG antibodies against human and bovine viruses. Diarrheal samples from calves were collected from a veterinary clinic and a commercial dairy farm in 2007 and 2008 (20). Written informed consent was obtained from parents of all children; the study was approved by the Institutional Review Board of the Christian Medical College, Vellore, India.The NoV GIII and NB VLPs were obtained from Linda Saif, Ohio State University (21). Validation assays were carried out prior to use of bovine VLPs using 20 bovine sera from a veterinary clinic. Goat anti-bovine IgG-horseradish peroxidase (IgG-HRP; (Jackson ImmunoResearch Inc., United States) was added, followed by addition of 3,3=,5,5=-tetramethylbenzidine substrate solution. The reaction was stopped with 2 M sulfuric acid after 15 min, and optical density (OD) was measured at 450 nm.Serum IgG was detected using plates coated overnight with 2 g/ml of human and bovine VLPs in phosphate-buffered saline (PBS) at 4°C, and the plates were blocked using 10% skim milk in PBS. Diluted serum samples were added to uncoated and VLPcoated wells and incubated. Anti-human IgG (Southern Biotech, United States) was added, followed by goat anti-mouse IgG-HRP (human adsorbed; Southern Biotech) and the substrate 2,2=-azino-di(3-ethylbenzthiazoline-6-sulfonate) solution; the reaction was stopped using 1% SDS solution, and OD was measured at 405 nm.The assays for human and bovine VLPs differed in the standards and controls included on each plate. The GI and GII standard curves were 2-fold dilutions of positive human sera starting at a 1:100 dilution, and the GI...

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confidence: 87%

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confidence: 98%

Exposure to Human and Bovine Noroviruses in a Birth Cohort in Southern India from 2002 to 2006

et al. 2013

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“…The production and purification of HS206 VLPs were performed as described previously (24). Briefly, Sf9 cells were infected with rBac-HS206 at a multiplicity of infection of 5 to 10.…”

Section: Methodsmentioning

confidence: 99%

“…The protein concentration of the VLPs was measured with Bradford reagent (Sigma-Aldrich, St. Louis, MO). The VLPs were negatively stained with 3% phosphotungstic acid (pH 7.0) and examined by transmission electron microscopy (EM) as described previously (24). The HS206 VLPs that were confirmed by EM to contain the correct morphologies of particles were used for hemagglutination (HA), saliva-VLP binding, and synthetic HBGA-VLP binding assays.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Emerging GII.g/GII.12 Noroviruses from a Gastroenteritis Outbreak in the United States in 2010

et al. 2011

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“…Subsequently, we cloned the capsid gene of HS66 and generated a recombinant baculovirus clone expressing the HS66 capsid gene. A baculovirus expression system was used to produce HS66 NoV VLPs using Spodoptera frugiperda (Sf9) insect cells (16). First, the capsid gene was amplified by RT-PCR from the original fecal sample RNA using forward primer HS66ORF2f (5Ј-GGCTCCCAGTTTTGTGAATG-3Ј) and reverse primer HS66ORF2r (5Ј-AACCAAGTCCAGAGCCAAGG-3Ј) with the following reaction characteristics: annealing temperature of 52°C and extension time of 2 min.…”

mentioning

confidence: 99%

Pathogenesis of a Genogroup II Human Norovirus in Gnotobiotic Pigs

Cheetham

Souza

Meulia

et al. 2006

J Virol

255

264

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Human noroviruses (HuNoVs) are a major cause of foodborne gastroenteritis worldwide. Because they do not grow in cell culture and there is no animal model for HuNoVs, pathogenesis studies have been hampered. Thus, little is known about their replication strategies or induction of neutralizing antibodies.The limited information on their pathogenesis is from human volunteer studies of HuNoV infections in which villus atrophy in duodenal biopsies and presence of malabsorptive diarrhea were described (1,7,8). No information is available on lesions in other portions of the intestines of these volunteers (9). Intestinal transplant pediatric patients that were diagnosed with HuNoV infection developed secretory or osmotic diarrhea (19,20,31). These patients had prolonged diarrhea (17 to 326 days) due to immunosuppressive therapy. The detection of HuNoV RNA and the clinical symptoms remitted after reduction of the immunosuppressive therapy. Usually in exposed individuals, histologic lesions correlate with diarrhea, but in one report, lesions in volunteers who did not show clinical symptoms were described (42). There are also numerous reports of asymptomatic individuals who were infected with HuNoVs and shed virus in the feces (11, 28).Most past attempts to study these viruses in an animal model may have failed because (i) the human strains that were used were not closely related to the host animal NoV strains, (ii) sensitive detection techniques were lacking, and finally (iii) the role of histo-blood group antigen (HBGA) phenotypes in differential susceptibility of the host was unrecognized. Our goal was to adapt a HuNoV strain to replicate in the gnotobiotic (Gn) pig to develop an animal model for the study of HuNoV pathogenesis. Gnotobiotic pigs are good models for human enteric diseases (40) because pigs resemble humans in their gastrointestinal anatomy, physiology, and immune responses. The Gn pigs are immunocompetent at birth, but they lack maternal antibodies and previous or ongoing exposure to microbial agents, including caliciviruses.Recently, viral RNA genetically similar to that of human NoV GII (65 to 71% amino acid sequence identity in the capsid gene) was detected in pigs in Japan (46, 47) and Europe (22,48). In U.S. swine, our laboratory detected both viral RNA and virus particles similar to GII HuNoV (70% sequence identity in the capsid region) which were infectious for Gn pigs (50). Our approach to infect Gn pigs with a HuNoV was to use a GII strain that is closely related genetically to the identified GII porcine NoVs and that has a broad HBGA binding pattern because little information or reagents are available for pig HBGA. Additionally, we used sensitive assays and reagents including reverse transcription (RT)-PCR to detect fecal shedding, virus-like particles (VLPs) for serological assays, and antisera to these VLPs for antigen enzyme-linked immunosor-

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Self-Assembly of the Recombinant Capsid Protein of a Bovine Norovirus (BoNV) into Virus-Like Particles and Evaluation of Cross-Reactivity of BoNV with Human Noroviruses

Cited by 24 publications

References 59 publications

Exposure to Human and Bovine Noroviruses in a Birth Cohort in Southern India from 2002 to 2006

Exposure to Human and Bovine Noroviruses in a Birth Cohort in Southern India from 2002 to 2006

Characterization of Emerging GII.g/GII.12 Noroviruses from a Gastroenteritis Outbreak in the United States in 2010

Pathogenesis of a Genogroup II Human Norovirus in Gnotobiotic Pigs

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