Human Caliciviruses Research Articles

Comparison of antigenic relationships between Hawaii and Mexican strains of Snow Mountain virus-like human caliciviruses using baculovirus-expressed viral capsid antigens 855

Comparison of antigenic relationships between Hawaii and Mexican strains of Snow Mountain virus-like human caliciviruses using baculovirus-expressed viral capsid antigens 855

Calicivirus emergence from ocean reservoirs: zoonotic and interspecies movements.

Caliciviral infections in humans, among the most common causes of viral-induced vomiting and diarrhea, are caused by the Norwalk group of small round structured viruses, the Sapporo caliciviruses, and the hepatitis E agent. Human caliciviruses have been resistant to in vitro cultivation, and direct study of their origins and reservoirs outside infected humans or water and foods (such as shellfish contaminated with human sewage) has been difficult. Modes of transmission, other than direct fecal-oral routes, are not well understood. In contrast, animal viruses found in ocean reservoirs, which make up a second calicivirus group, can be cultivated in vitro. These viruses can emerge and infect terrestrial hosts, including humans. This article reviews the history of animal caliciviruses, their eventual recognition as zoonotic agents, and their potential usefulness as a predictive model for noncultivatable human and other animal caliciviruses (e.g., those seen in association with rabbit hemorrhagic disease).

Prevalence of antibodies to the Hawaii strain of human calicivirus as measured by a recombinant protein based immunoassay

The evaluation of an enzyme immunoassay using recombinant Hawaii virus-like particles (rHV-LPs) with a panel of sera which had been screened previously for antibodies to Norwalk virus (NV) and Mexico virus (MxV) is described. The assay was also applied to study the epidemiology of Hawaii virus. Adult volunteers challenged with the prototype (genogroup II, human calicivirus) HV developed significant IgG responses (16-32 fold rises) following challenge whereas adults challenged or naturally infected with NV (genogroup I) did not. Lesser antibody responses (4-8 fold rises) were demonstrated in volunteers challenged with Snow Mountain agent (SMA) and patients infected by SRSV 'UK3' and 'UK4' strains, indicating a degree of antigenic relatedness among viruses within genogroup II. Comparison of the seroprevalence of IgG antibodies to rHV, rMxV and rNV in 338 children in London showed that infections with genogroup II viruses are prevalent and occur earlier in life than NV. Many young children had antibodies to MxV but not HV indicating that genogroup II viruses have both conserved and antigenically distinct epitopes. A serological study on 566 Canadians aged between 9 and 79 years showed that the prevalence of antibodies to rHV rose with age from 65-100% and from 53-100% for NV. Measurement of antibody response in a heart transplant patient infected with an MxV-like virus showed significant responses to both rMxV and rHV. Continuous monitoring of the patient over two years showed that antibody levels declined rapidly to prechallenge levels after a year.

Prevalence of antibodies to the Hawaii strain of human calicivirus as measured by a recombinant protein based immunoassay

The evaluation of an enzyme immunoassay using recombinant Hawaii virus–like particles (rHVLPs) with a panel of sera which had been screened previously for antibodies to Norwalk virus (NV) and Mexico virus (MxV) is described. The assay was also applied to study the epidemiology of Hawaii virus. Adult volunteers challenged with the prototype (genogroup II, human calicivirus) HV developed significant IgG responses (16–32 fold rises) following challenge whereas adults challenged or naturally infected with NV (genogroup I) did not. Lesser antibody responses (4–8 fold rises) were demonstrated in volunteers challenged with Snow Mountain agent (SMA) and patients infected by SRSV ‘UK3’ and ‘UK4’ strains, indicating a degree of antigenic relatedness among viruses within genogroup II. Comparison of the seroprevalence of Ig G antibodies to rHV, rMxV and rNV in 338 children in London showed that infections with genogroup II viruses are prevalent and occur earlier in life than NV. Many young children had antibodies to MxV but not HV indicating that genogroup II viruses have both conserved and antigenically distinct epitopes. A serological study on 566 Canadians aged between 9 and 79 years showed that the prevalence of antibodies to rHV rose with age from 65–100% and from 53–100% for NV. Measurement of antibody response in a heart transplant patient infected with an MxV-like virus showed significant responses to both rMxV and rHV. Continuous monitoring of the patient over two years showed that antibody levels declined rapidly to prechallenge levels after a year. J. Med. Virol. 54:135–139, 1998. © 1998 Wiley-Liss,Inc.

Detection of Norwalk Virus and Other Genogroup 1 Human Caliciviruses by a Monoclonal Antibody, RecombinantAntigen-Based Immunoglobulin M Capture Enzyme Immunoassay

Detection of Norwalk Virus and Other Genogroup 1 Human Caliciviruses by a Monoclonal Antibody, RecombinantAntigen-Based Immunoglobulin M Capture Enzyme Immunoassay

Sapporo-like human caliciviruses are genetically and antigenically diverse.

The Sapporo-like human caliciviruses (HuCVs) comprise one of three genogroups of HuCVs associated with acute gastroenteritis. Phylogenetic analysis has shown that Sapporo-like HuCVs are related more closely to animal caliciviruses than to other known HuCVs. We produced 3.2 kb cDNA fragments from the 3' end to three Sapporo-like HuCVs that were associated with acute gastroenteritis in children (Houston/86, Houston/90, and London/92). Sequence analysis of the 3.2 kb cDNAs showed that two of the three viruses had a genomic organization similar to that of other Sapporo-like strains and the third strain (London/92) lacked an open reading frame overlapping the 5' end of the capsid gene. Alignment of the capsid sequences of these three strains showed 44-78% amino acid identity among the three strains. Phylogenetic analysis of the aligned sequences indicated the three strains are related but each belongs to a distinct genetic cluster. The genetic differences are associated with antigenic differences in that an enzyme immune assay (EIA) specific for the prototype Sapporo/82 strain detected the Houston/86 strain, but not the Houston/90 and London/92 strains. In vitro transcription and translation of viral cDNA containing the predicted capsid gene of Houston/90 resulted in a protein of 63 K, which is immunoprecipitated by sera from children infected with the strain. Genetically and antigenically distinct strains in the Sapporo-like HuCVs have not been described previously and the occurrence of such diverse strains in the same community likely increases the importance of these strains as a cause of illness in children.

Molecular characterization of morphologically typical human calicivirus Sapporo

Human calicivirus Sapporo (SV) has typical calicivirus morphology and causes acute gastroenteritis in children. The nucleotide sequence of 3.2 kb of the 3' end of SV was determined from a cloned cDNA. The 3' end of the SV genome is predicted to encode the RNA-dependent RNA polymerase region, the capsid protein and two small open reading frames. The nonstructural and capsid protein coding sequences in the SV genome are fused in a single open reading frame. The organization of these proteins in the SV sequence is similar to that of rabbit hemorrhagic disease virus and the recently described Manchester virus, and distinct from the genome organization of the prototype human calicivirus, Norwalk virus, that lacks typical calicivirus morphology and has been described as a small round structured virus (SRSV). Sequence analysis of the predicted capsid region showed that the SV capsid is longer by approximately 30 amino acids than the capsid of any of the SRSVs, and multiple sequence alignments showed that these additional amino acids are located in the variable region of the capsid protein. Expression of the capsid protein of SV in insect cells resulted in the self-assembly of virus-like particles that have a morphology similar to that of the native virus. This result shows that calicivirus morphology is determined by the primary sequence of the capsid protein.

Phylogenetic analysis of the Caliciviruses.

A phylogenetic portrait of the genus Calicivirus in the family Caliciviridae was developed based upon published sequences and newly characterized calicivirus (CV) strains, including additional Sapporo-like HuCV strains in pediatric diarrhea stool specimens from South Africa, the United Kingdom, and the United States. Distance and parsimony methods were applied to nucleotide and amino acid sequences of human and animal calicivirus 3D RNA-dependent RNA polymerase (approximately 470nt) and capsid hypervariable regions (approximately 1,200nt) to generate phylogenetic trees. Pairwise amino acid identity in the 3D region among the Sapporo-like strains ranged from 61% to 100%. Human and animal caliciviruses (HuCVs and AnCVs) separated into five genogroups: small round-structured viruses (SRSV), Sapporo-like, and hepatitis E virus (HEV)-like HuCVs and rabbit-, and vesicular exanthema of swine virus (VESV)-like AnCVs, each with a distinct genome organization. Each genogroup, including the Sapporo-like HuCVs, subdivided further into subgenogroups. The capsid region trees had higher levels of confidence than the 3D region trees and limited conclusions about genogroups could be drawn from the 3D region analyses. This analysis suggested that CVs include five potential virus subfamilies.

Morphologic Identification of Viral Gastroenteritis Caused by a Human Calicivirus Strain Undetected Initially by Molecular or Immunologic Methods

Molecular and immunologie assays for virus identification and characterization are well known for their high specificity and sensitivity. High specificity occasionally hampers efforts to detect virus strains distantly related to previously recognized types and may allow infectious agents to be undetected when these techniques are used for identification. We describe in this report such an example with an outbreak of foodborne gastroenteritis.Forty-six cases of gastrointestinal illness were reported in May 1994 among adult staff of a Parkville, Md., school after a catered luncheon. Symptoms included stomach cramps, nausea, diarrhea, and vomiting. Seven stool specimens, negative by routine bacteriological screening, and 14 acute-and convalescent-phase sera from patients involved in the outbreak were tested for viruses. The stool specimens were examined by electron microscopy (EM) and reverse transcription-polymerase chain reaction (RT-PCR) designed to detect small round structured viruses (SRSVs). In addition, serum specimens were tested for IgG antibody to recombinant capsid proteins from Norwalk virus (rNV) and Toronto virus (rTV) using a direct enzyme immunoassay (EIA).

Human calicivirus genogroup II capsid sequence diversity revealed by analyses of the prototype Snow Mountain agent.

The Snow Mountain agent (SMA) is the prototype genogroup II and serotype 3 human calicivirus responsible for epidemic outbreaks of acute gastroenteritis. We have cloned the region of the SMA genome that encodes the single capsid protein. The predicted amino acid sequence of the capsid protein is distinct from other calicivirus strains that have been termed SMA-like based on sequence similarity between the RNA polymerase regions and IEM reactivity. In a previous report, a high sequence similarity in a small region of the RNA polymerase between SMA and another strain, OTH-25, suggested that the capsid proteins of OTH-25 and SMA would be very similar. In this report, we show that the capsid proteins of OTH-25 and SMA are more distinct than was predicted by similarity in the RNA polymerase. In addition, phylogenetic analysis of a region of the RNA polymerase and of the N-terminal conserved domain of the capsid protein of 12 human caliciviruses resulted in trees with different topologies, suggesting that recombination has occurred within this group of viruses. Molecular characterization of the prototype calicivirus strains is important in determining the relationships between capsid similarity at the amino acid level, genetic grouping by sequence comparison, and antigenic reactivity.

Expression and self-assembly of recombinant capsid protein from the antigenically distinct Hawaii human calicivirus.

The Norwalk and Hawaii viruses are antigenically distinct members of the family Caliciviridae and are considered to be important etiologic agents of epidemic gastroenteritis, with most studies focusing on the role of Norwalk virus. To further investigate the importance of Hawaii virus, Hawaii virus-like particles (VLPs) were produced by expression of its capsid protein in the baculovirus system and these VLPs were used as the antigen in an enzyme-linked immunosorbent assay that was efficient in the detection of a serologic response to Hawaii virus. The ready availability of Hawaii VLPs should enable larger-scale epidemiological studies to further elucidate the importance of this agent.

Parkville virus: A novel genetic variant of human calicivirus in the Sapporo virus clade, associated with an outbreak of gastroenteritis in adults

Parkville virus: A novel genetic variant of human calicivirus in the Sapporo virus clade, associated with an outbreak of gastroenteritis in adults

Parkville virus: A novel genetic variant of human calicivirus in the Sapporo virus clade, associated with an outbreak of gastroenteritis in adults

This report describes the characterization of Parkville virus, the etiologic agent of an outbreak of foodborne gastroenteritis, that has the morphology of a calicivirus and genetic properties that distinguish it from previously identified strains in the Sapporo/Manchester virus clade. Sequence analysis of the Parkville virus genome showed it contained the RNA-dependent RNA polymerase motifs GLPSG and YGDD characteristic of members of the family Caliciviridae with an organization identical to that reported for the Manchester virus where the capsid region of the polyprotein is fused to the RNA polymerase. Parkville virus however, demonstrates considerable sequence divergence from both the Manchester and Sapporo caliciviruses, providing the first indications that genetic diversity exists within caliciviruses of this previously homogeneous clade. On the basis of recent advances in the genetic characterization of members of the family Caliciviridae, we propose a new interim phylogenetic classification system in which Parkville virus would be included with Manchester and Sapporo virus as a separate group distinct from the small round-structured viruses (Norwalk-like viruses) that also cause diarrhea in humans.

Incidence of human calicivirus and rotavirus infection in patients with gastroenteritis in South Africa.

Human caliciviruses (HuCVs) are reportedly responsible for 2.5-4% of nonbacterial sporadic gastroenteritis. The incidence of HuCV infection in South Africa is unknown. Stool specimens from 1,296 South African patients with sporadic gastroenteritis were screened for the presence of HuCVs using electron microscopy, recombinant enzyme immunoassays for Norwalk (NV) and Mexican (MX) viruses, and the reverse transcriptase-polymerase chain reaction (RT-PCR). RT-PCR products were sequenced to ascertain which HuCV genogroups were present. HuCVs were detected in 43/1,296 (3.3%) specimens examined, with RT-PCR proving to be the most sensitive detection method. Genetic analysis of the isolates indicated that 81% were Snow Mountain Agent, or MX-like; 8% were NV-like; and 11% were HuCV/Sapporo-like. This study indicates that a combination of assays is needed for the accurate detection of HuCVs. Comparative data on hospitalised patients showed that the incidence of rotavirus infection was approximately ten times greater than that of HuCV infection.

Virologic Features of Sapporo-like Human Calicivirus Infections During an Outbreak in a Houston Child Care Center 432

Virologic Features of Sapporo-like Human Calicivirus Infections During an Outbreak in a Houston Child Care Center 432

Prevalence of antibodies to human caliciviruses (HuCVs) in Kuwait established by ELISA using baculovirus‐expressed capsid antigens representing two genogroups of HuCVs

Baculovirus recombinant-expressed antigens of Norwalk viruses (rNV) and a Mexico strain (rMX) of the Snow Mountain serogroup of human caliciviruses (HuCVs) were used in enzyme immunoassays to study the antibody prevalence among the Kuwaiti population and foreign workers employed in Kuwait. The antibody titers in 16 different age groups which ranged from neonates to centenarians were investigated by testing eight different dilutions of each serum (1:200–1:25,600). The results indicate that NV infection is widespread in Kuwait and affects all age groups. Ninety-eight percent of the 433 serum samples tested had antibodies to rNV. In the 50–79-year-old age group, the antibody levels to rNV were higher and significantly different from those in children 0–7 years old. In infants, the rNV antibodies did not diminish by 4 months of age and their titer steadily increased with age. When 414 of these sera samples were tested for antibodies to rMX, 96% positive serological responses were observed. Antibody titers to rMX were reduced in infants from 4 to 11 months; however, 95% of the samples were positive. These data indicate that children born in Kuwait are infected with Norwalk-like viruses at a very early age. Finally, antibodies to rNV and rMX were found in 98% of 151 and in 95% of 148 foreign workers, respectively. J Med Virol 51:115–118, 1997. © 1997 Wiley-Liss, Inc.

Prevalence of antibodies to human caliciviruses (HuCVs) in Kuwait established by ELISA using baculovirus-expressed capsid antigens representing two genogroups of HuCVs

Baculovirus recombinant-expressed antigens of Norwalk viruses (rNV) and a Mexico strain (rMX) of the Snow Mountain serogroup of human caliciviruses (HuCVs) were used in enzyme immunoassays to study the antibody prevalence among the Kuwaiti population and foreign workers employed in Kuwait. The antibody titers in 16 different age groups which ranged from neonates to centenarians were investigated by testing eight different dilutions of each serum (1:200-1:25,600). The results indicate that NV infection is widespread in Kuwait and affects all age groups Ninety-eight percent of the 433 serum samples tested had antibodies to rNV. In the 50-79-year, old age group, the antibody levels to rNV were higher and significantly different from those in children 0-7 years old. In infants, the rNV antibodies did not diminish by 4 months of age and their titer steadily increased with age. When 414 of these sera samples were tested for antibodies to rMX, 96% positive serological responses were observed. Antibody titers to rMX were reduced in infants from 4 to 11 months; however, 95% of the samples were positive. These data indicate that children born in Kuwait are infected with Norwalk-like viruses at a very early age. Finally, antibodies to rNV and rMX were found in 98% of 151 and in 95% of 148 foreign workers, respectively.

Viral zoonoses and food of animal origin: caliciviruses and human disease.

Caliciviruses are important veterinary and human pathogens. The viruses gain their name from characteristic cup-shaped structures seen on the virion surface by negative stain electron microscopy. In humans caliciviruses are a major cause of diarrhoeal disease. There are two fundamentally different genome structures amongst human caliciviruses. The Norwalk-like or small round structured viruses (SRSVs) are viruses that have an amorphous structure when viewed by EM, they have a genome composed of 3 major open reading frames (ORFs). These viruses cause epidemic gastroenteritis amongst all age groups. In contrast, the 'classic' human caliciviruses (HuCVs) display the typical calicivirus surface structure and have their capsid ORF fused to and contiguous with the non structural proteins forming one giant polyprotein. HuCVs are predominantly associated with paediatric infections and are only a minor cause of disease in humans. Spread of disease for both SRSVs and HuCVs is usually by faecal oral transmission. SRSVs are a major cause of foodborne gastroenteritis especially linked to the consumption of sewage-contaminated shellfish. However, there is no evidence that these viruses replicate in shellfish or that they originate from an animal source.

Sequence analysis of the gene encoding the capsid protein of the Snow Mountain human calicivirus.

Snow Mountain virus (SMV) is the reference strain for serotype 3 as determined by immune electron microscopy of the human caliciviruses that are associated with epidemic gastroenteritis. In order to establish the genetic relationship of its capsid protein with those from other human caliciviriuses, the sequence of the open reading frame 2 (ORF2) encoding the SMV capsid protein was determined. The SMV ORF2 sequence was 1626 nucleotides in length and the deduced protein of 542 amino acids had a calculated molecular weight of 59.2 kD. The SMV capsid sequence showed approximately 48 and 77% amino acid sequence identity with the capsid proteins of the Norwalk (serotype 1) and Hawaii (serotype 2) human calicivirus reference strains, respectively, a finding consistent with its serotypic distinctiveness. Furthermore, the predicted amino acid sequence of the SMV capsid was found to share highest sequence identity (98%) with the Melksham human calicivirus in database searches.

Viral infections of the gastrointestinal tract

Viral gastroenteritis is an important global problem that is being actively investigated. This year, stimulating new concepts regarding pathogenesis and immunity of the major viral pathogen of the gastrointestinal tract, rotavirus, were reported. In addition, highly promising results from phase 3 viral vaccine trials suggest that an effective vaccine for rotavirus will be available in the next several years. The range and diversity of human caliciviruses continues to be defined as more isolates are characterized. The ability to produce recombinant virus-like particles of human caliciviruses has enhanced diagnostic capability and enabled more extensive epidemiologic studies. The roles that human astrovirus, adenovirus, and newer agents, such as Aichi virus, play in diarrheal disease continue to be investigated.