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At least 217 records · Page 12Linked to original sources

PubMed1998

Rotavirus 2/6 viruslike particles administered intranasally with cholera toxin, Escherichia coli heat-labile toxin (LT), and LT-R192G induce protection from rotavirus challenge.

We have shown that rotavirus 2/6 viruslike particles composed of proteins VP2 and VP6 (2/6-VLPs) administered to mice intranasally with cholera toxin (CT) induced protection from rotavirus challenge, as measured by virus shedding. Since it is unclear if CT will be approved for human use, we evaluated the adjuvanticity of Escherichia coli heat-labile toxin (LT) and LT-R192G. Mice were inoculated intranasally with 10 microg of 2/6-VLPs combined with CT, LT, or LT-R192G. All three adjuvants induced equivalent geometric mean titers of rotavirus-specific serum antibody and intestinal immunoglobulin G (IgG). Mice inoculated with 2/6-VLPs with LT produced significantly higher titers of intestinal IgA than mice given CT as the adjuvant. All mice inoculated with 2/6-VLPs mixed with LT and LT-R192G were totally protected (100%) from rotavirus challenge, while mice inoculated with 2/6-VLPs mixed with CT showed a mean 91% protection from challenge. The availability of a safe, effective mucosal adjuvant such as LT-R192G will increase the practicality of administering recombinant vaccines mucosally.

Administration, Intranasal↗

PubMed2005

Viremia and nasal and rectal shedding of rotavirus in gnotobiotic pigs inoculated with Wa human rotavirus.

Respiratory symptoms with rotavirus shedding in nasopharyngeal secretions have been reported in children with and without gastrointestinal symptoms (Zheng et al., 1991, J. Med. Virol. 34:29-37). To investigate if attenuated and virulent human rotavirus (HRV) strains cause upper respiratory tract infections or viremia in gnotobiotic pigs, we inoculated them with attenuated or virulent HRV intranasally, intravenously, or orally or via feeding tube (gavage) and assayed virus shedding. After oral or intranasal inoculation with attenuated HRV, the pigs remained asymptomatic, but 79 to 95% shed virus nasally and 5 to 17% shed virus rectally. After inoculation by gavage, no pigs shed virus nasally or rectally, but all pigs seroconverted with antibodies to HRV. No viremia was detected through postinoculation day 10. Controls inoculated intranasally with nonreplicating rotavirus-like particles or mock inoculated did not shed virus. In contrast, 100% of pigs inoculated with virulent HRV (oral, intranasal, or gavage) developed diarrhea, shed virus nasally and rectally, and had viremia. The infectivity of sera from the viremic virulent HRV-inoculated pigs was confirmed by inoculating gnotobiotic pigs orally with pooled HRV-positive serum. Serum-inoculated pigs developed diarrhea and fecal and nasal virus shedding and seroconverted with serum and intestinal HRV antibodies. Pigs inoculated intravenously with serum or intestinal contents from the viremic virulent HRV-inoculated pigs developed diarrhea, virus shedding, and viremia, similar to the orally inoculated pigs. This study provides new evidence that virulent HRV causes transient viremia and upper respiratory tract infection in addition to gastrointestinal infection in gnotobiotic pigs, confirming previous reports of rotavirus antigenemia (Blutt et al., Lancet 362:1445-1449, 2003). Our data also suggest that intestinal infection might be initiated from the basolateral side of the epithelial cells via viremia. Additionally, virus shedding patterns indicate a different pathogenesis for attenuated versus virulent HRV.

Animals↗

PubMed2006

Pentavalent human-bovine reassortant rotavirus vaccine: a review of its efficacy and safety in preventing acute rotavirus gastroenteritis in healthy infants.

Acute rotavirus gastroenteritis can be successfully managed with oral and intravenous rehydration solutions; however, it is still a major cause of childhood mortality and morbidity in developing countries and is responsible for significant morbidity and health-care costs in developed countries. Pentavalent human-bovine rotavirus vaccine (HBRV) is an oral vaccine containing five live attenuated human-bovine reassortant strains containing the human serotypes responsible for the majority of rotavirus cases worldwide. Approximately three-quarters of all acute rotavirus gastroenteritis cases and almost all cases of severe gastroenteritis were prevented by pentavalent HBRV in a phase III clinical trial. Pentavalent HBRV has a favorable safety profile, with fever, irritability, vomiting and diarrhea generally reported in similar proportions of infants who received either vaccine or placebo. Results from a large phase III study indicate that there was no excess risk of intussusception among those vaccinated compared with placebo recipients. This review summarizes the current data on the clinical efficacy and safety of HBRV in healthy infants.

Gastroenteritis↗

PubMed1999

[The molecular epidemiology of the rotavirus in Spanish children. The Rotavirus Study Group (GER)].

BACKGROUND: Rotavirus are the most common etiologic agent of acute gastroenteritis in childhood. The knowledge of the circulating antigenic types is important in development of future vacunes. METHODS: Faeces from children (age < 4 years) with acute gastroenteritis admitted in the two hospitals (Hospital Severo Ochoa-Madrid and Hospital General Vic-Barcelona) have been studied prospectively during one year (October-1996 to October-1997). The detection of rotavirus was performed by ELISA (IDEIA, Dako). All samples were G-serotyping by EIA-Mabs (Silenius Laboratories) and the indeterminate or non-serotypable samples were G-genotyping by RT-PCR. P genotypes were identified by RT-PCR. RESULTS: 322 (45%) patients with acute diarrhoea causing for rotavirus were confirmed, 242 coming from the Madrid metropolitan area and other 80 from the Barcelona area. The EIA-Mabs technique made it possible to identify the G serotypes in 287 cases (89%), corresponding 207 to G1 serotype, 70 to G4 serotype and 6 to G3 serotype. In 4 patients both G1 and G4 serotypes were detected. The EIA-Mabs could not determined the serotype in 35 (11%) patients, all of whom were confirmed by RT-PCR (12 belonged to serotype G1 and 23 to serotype G4). Analysis of P genotypes was carried out in 25 patients obtained from Madrid and 17 from Barcelona; all cases were classified in the P[8] genotype. CONCLUSIONS: The most frequent serotype in both hospitals was G1. The EIA-Mabs technique were showed a high sensitivity, however, the RT-PCR technique used were even more efficient, making it possible for us to identify all the non-serotypable EIA-Mabs cases. The temporal study of circulating serotypes/genotypes of rotavirus is necessary to evaluate the efficiency of vaccines.

Acute Disease↗

PubMed1992

Production of monoclonal antibodies to rotavirus suitable for detection of rotavirus in stool samples by one-step EIA.

Two stable hybridoma cell clones producing high amounts of monoclonal antibodies (MAb) to rotavirus have been selected. The MAbs were shown to recognize different epitopes of the major inner capsid rotavirus protein VP 6. The two types of MAb in question cross-reacted with human and animal group A rotaviruses. Due to their high affinity and good binding capacity to microtiter plates a simultaneous EIA was developed for detection of rotavirus in stool samples. The sensitivity and specificity of MAb one-step EIA was compared with an approved polyclonal sandwich EIA by testing 1309 stool samples.

Animals↗

PubMedDate not supplied

Demonstration of rotavirus and rotavirus-like virus in the intestinal contents of diarrheic pheasant chicks.

Intestinal content samples from four flocks of pheasant chicks experiencing diarrhea and increased mortality were evaluated using immune electron microscopy and genome electropherotyping techniques. Enteric viruses serologically and electropherotypically indistinguishable from turkey rotaviruses and turkey rotavirus-like viruses (RVLVs) were detected. Rotavirus-only infections, RVLV-only infections, and simultaneous rotavirus and RVLV infections were demonstrated in these flocks.

Animals↗

PubMed1984

Waterborne outbreak of rotavirus diarrhoea in adults in China caused by a novel rotavirus.

During two epidemics of acute diarrhoea in China in late 1982/early 1983, more than 12 000 adults in two coal mining districts were affected. The virus isolated from stool samples resembled a rotavirus morphologically. Antigenically it lacked the group antigen shared by known rotaviruses. Like other rotaviruses it had a double-stranded RNA with 11 discrete segments, but the pattern of migration of the segments on polyacrylamide electrophoresis differed from those of other rotaviruses.

Adolescent↗

PubMedDate not supplied

[The determination of human IgM rotavirus antibodies in the blood serum by an immunoenzyme method in patients with a rotavirus infection].

Rotavirus infection is one of the infectious diseases which can be verified only by a laboratory assay. That is why elaboration of new methods of the infection serodiagnosis is of great importance. Enzyme-linked immunoassay was worked up for IgM rotavirus antibodies detection in human blood serum. Validation of the method provides grounds for its being recommended for the diagnosis of rotavirus infection alongside with the determination of rotavirus antigens in feces.

Antibodies, Viral↗

PubMed1994

Evaluation of the antigenicity and reactogenicity of varying formulations of the rhesus rotavirus-based quadrivalent and the M37 rotavirus vaccine candidates.

Three phase I trials of the rhesus rotavirus (RRV)-based quadrivalent vaccine [composed of serotype 3 (RRV), and serotypes 1 (D x RRV), 2 (DS1 x RRV), and 4 (ST3 x RRV) human rotavirus x RRV reassortants] and the M37 (nursery strain) rotavirus vaccine candidates were conducted in an attempt to find a safe and optimally antigenic formulation. Infants 10-20 weeks old received, in trial I, 1) the quadrivalent vaccine as two separate bivalent doses (1 x 10(4) PFU each of D x RRV and RRV, followed 4 weeks later by 1 x 10(4) PFU each of DS1 x RRV and ST3 x RRV) or 2) placebo; in trial II, 1) one dose of quadrivalent vaccine (10(4) PFU of each component), or 2) two doses of quadrivalent vaccine, or 3) a 10(4) PFU dose of M37 vaccine, or 4) M37 vaccine followed by the quadrivalent vaccine, or 5) placebo; in trial III, 1) a dose of a higher-titered quadrivalent vaccine (10(5) PFU of each component), or 2) two doses of higher titered quadrivalent vaccine, or 3) a dose of higher titered M37 vaccine (10(5) PFU) or 4) two doses of M37 vaccine (10(5) PFU), or 5) M37 vaccine (10(5) PFU) followed by the higher titered quadrivalent vaccine, or 6) placebo. A mild, transient fever during the first week postvaccination was associated with the bivalent or quadrivalent vaccines but not with the M37 vaccine. Fourfold or greater serum IgA ELISA responses to rotavirus were observed in 48-92% of the infants receiving quadrivalent vaccine and in 32-50% of those receiving M37 vaccine.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Viral↗

PubMed1993

Both surface proteins (VP4 and VP7) of an asymptomatic neonatal rotavirus strain (I321) have high levels of sequence identity with the homologous proteins of a serotype 10 bovine rotavirus.

The nucleotide sequence of genes 4 and 9, encoding the outer capsid proteins VP4 and VP7 of a serotype 10 tissue culture-adapted strain, I321, representative of asymptomatic neonatal rotaviruses isolated from neonates in Bangalore, India, were determined. Comparison of nucleotide and deduced amino acid sequences of I321 VP4 and VP7 with previously published sequences of various serotypes revealed that both genes were highly homologous to the respective genes of serotype 10 bovine rotavirus, B223. The VP4 of I321 represents a new human P serotype and the I321 and related strains represent the first description of neonatal rotaviruses that appear to derive both surface proteins from an animal rotavirus.

Amino Acid Sequence↗

PubMed1994

Expression and sequence analysis of gene 7 of the IDIR agent (group B rotavirus): similarity with NS53 of group A rotavirus.

The seventh genomic segment of the IDIR strain of group B rotavirus was cloned, sequenced, and expressed in vitro in order to evaluate the gene coding assignment by comparison with group A rotaviruses (GAR). Viral genomic RNA for these reactions was obtained directly from fecal specimens of infected infant rats. IDIR virus gene 7 (IDIRg7) contained 1276 bp. Both 5' and 3' termini resembled those reported for other IDIR virus genomic segments. Two open reading frames (ORF) were predicted from the gene sequence: a long ORF from bases 258-1217 and a short ORF from bases 41-385. The first AUG of the long ORF was flanked by sequences associated with highly efficient translation, but less efficient translation was predicted for the short ORF. In vitro transcription and translation of IDIRg7 demonstrated a product consistent with polypeptide synthesis from the long ORF but not the short ORF. Convalescent rat antibody directed against the IDIR agent specifically immunoprecipitated the IDIRg7 protein and indicated that the in vitro translation product was indeed encoded by the virus genome. Thus, the untranslated 5' region of IDIRg7 was longer than that previously described for any major rotavirus product. Comparison of the IDIRg7 sequence indicated 51% similarity and 18% identity with the amino acid residues of NS53 of the SA11 strain of GAR. However, the sequence of IDIRg7 did not share the putative zinc binding region postulated for NS53 of rotavirus groups A and C. It will be of interest in future experiments to evaluate the function of the IDIRg7 product following large-scale synthesis by alternative expression systems such as vaccinia or baculovirus.

Amino Acid Sequence↗

PubMed1979

Simian rotavirus (SA 11) in serodiagnosis of human rotavirus infections.

Tests were made on 169 sera from children up to 10 years of age for rotavirus antibodies by indirect immunofluorescence and complement fixation tests. The simian strain SA 11 served as a substitute antigen for the human rotavirus. Furthermore, the SA 11 complement-fixing antigen was compared with a commercial antigen of the bovine rotavirus strain NCDV. It was demonstrated that SA 11 which can be more easily propagated than most bovine strains, may be used as a substitute antigen in complement fixation and indirect immunofluorescence for serodiagnosis of human rotavirus infections.

Animals↗

PubMed1997

Comparative amino acid sequence analysis of the outer capsid protein VP4 from four lapine rotavirus strains reveals identity with genotype P[14] human rotaviruses.

The genes encoding the outer capsid VP4 proteins of four lapine rotavirus strains, three isolated in the US (ALA, C-11 and BAP-2) and one isolated in Japan (R-2) were sequenced, and the predicted amino acid (aa) sequence was compared to all known rotavirus genotypes. A high degree of aa identity (96.8-98.9%) was found among the American lapine strains, while the Japanese rotavirus strain R-2 shared less aa identity (89.5-90.0%) with the American strains. The four lapine rotaviruses shared the closest aa identity (90.6-94.9%) with the P[14] genotype, consisting of viruses isolated from humans in Italy, Finland and Thailand. These results indicate that the VP4 protein of the four lapine strains are genotype P14, and that among lapine strains there are possibly two subtypes, one represented by the American lapine strains and the other by the Japanese R-2 strain.

Amino Acid Sequence↗

PubMed1991

Molecular analysis of the gene 6 from a porcine group C rotavirus that encodes the NS34 equivalent of group A rotaviruses.

The complete nucleotide sequence of the gene 6 from the porcine group (Gp) C rotavirus strain Cowden was determined from gene 6-specific clones selected from a cDNA library and from viral transcript RNA. The gene is 1348 nucleotides in length with a potential initiation codon beginning at nucleotide 25 and a stop codon at nucleotide 1231. The deduced protein contains 402 amino acids. Comparison of the gene 6 from this Gp C strain with sequences in the GenBank data base indicated that this gene shared homology with gene 7 of Gp A rotavirus strain SA-11 (22.9%) and gene 9 of Gp A rotavirus strain UK (22.6%), both of which encode the NS34 protein. In vitro translation products produced from transcripts generated from a gene 6 clone containing the entire open reading frame were not immunoprecipitated with either hyperimmune serum specific for the Gp C Cowden strain or a monoclonal antibody directed against the group antigen (VP6) of the Cowden strain. However, products generated from a full-length gene 5 clone of the Cowden strain were immunoprecipitated by each of these antibodies. These data suggest that in contrast to the Gp A viruses in which the gene 6 encodes the major inner capsid protein VP6, the gene 6 of the Cowden Gp C strain encodes a nonstructural protein corresponding to the NS34 of Gp A rotaviruses.

Amino Acid Sequence↗

PubMed1992

Sequences of the four larger proteins of a porcine group C rotavirus and comparison with the equivalent group A rotavirus proteins.

The sequences of the four larger proteins of rotavirus group C (Cowden strain) are presented and compared with the sequences of the corresponding group A proteins. They exhibit a significant level of homology, allowing gene coding assignment for the group C rotavirus. The coding strategy of the group C virus RNA segment is the same as that for the group A large segments as one long open reading frame is present in each segment. The genome segment 1 encodes the structural protein VP1 which presents the RNA-dependent RNA polymerase consensus motifs. The VP1 protein is the most highly conserved between the rotaviruses of groups A and C. The genome segment 2 encodes the VP2 protein. The deduced protein sequence does not present the putative leucine zippers identified in the group A protein but its amino terminal is hydrophilic and highly charged as previously noted for the group A protein. The genome segment 3 encodes for a protein homologous to the group A outer capsid protein VP4. As observed among the various group A sequences, the amino terminal is the region presenting the fewest similarities. A cleavage region and a putative fusion motif similar to those present in the group A viruses have been identified. For this protein the comparison has been extended to the IDIRV [corrected] VP3 previously sequenced and indicates that groups A and C VP4 proteins are much more related to each other than to the group B equivalent. The genome segment 4 encodes for a protein showing an approximate 40% sequence identity to the minor core protein, VP3, of the group A rotavirus. This remarkable conservation of primary structures argues for severe functional constraint on the evolution of these proteins.

Amino Acid Sequence↗

PubMed1985

Sequence of the serotype-specific glycoprotein of the human rotavirus Wa strain and comparison with other human rotavirus serotypes.

Complementary DNA was synthesized from the double-stranded RNA of the Wa strain of human rotavirus and inserted into the bacterial plasmid pBR322. Clones which contained the gene that codes for the viral glycoprotein (VP7) were identified and the nucleotide sequence was determined. The gene was 1062 base pairs in length with an open reading frame which coded for 326 amino acids. Two potential glycosylation sites were found as well as two hydrophobic regions at the N-terminus of the polypeptide. The untranslated regions at the 5' and 3' ends were 48 base pairs and 33 base pairs long, respectively. Only one nucleotide at position 493 differed from the sequence of the Wa VP7 gene described by Richardson et al. (1984, J. Virol. 51, 860-862). A strong prokaryotic promoter sequence was also found between residues 434 and 462. A comparison of the amino acid sequence of the Wa strain (serotype 1) to the Hu/5 strain of human rotavirus (serotype 2) and SA11, the simian rotavirus (serotype 3), revealed a high degree of homology (79.1% and 83.1%, respectively) between the serotypes, suggesting that rotavirus serotypes are stable. The hydrophilic regions of VP7 of the three serotypes were identified and compared for homology. Four of these regions showed variation between serotypes.

Amino Acid Sequence↗

PubMed1998

Passive immunity to bovine rotavirus in newborn calves fed colostrum supplements from cows immunized with recombinant SA11 rotavirus core-like particle (CLP) or virus-like particle (VLP) vaccines.

Heterotypic passive immunity to IND (P/5/G6) bovine rotavirus (BRV) was evaluated. Three groups of calves (n = 5 per group) were fed 1% pooled colostrum supplements (birth to 7 days of age) from BRV seropositive cows vaccinated with recombinant SA11(P/2/G3) rotavirus-like particles (VLPs), recombinant SA11 rotavirus core-like particles (CLPs), or inactivated SA11 rotavirus (SA11). Control calves (n = 5 per group) received either pooled colostrum from unvaccinated (BRV field exposure seropositive) control cows, or no colostrum. IgG1 antibody titers to IND BRV for the pooled colostrum were: 1,048,576 (VLP); 1,048,576 (CLP); 262,144 (SA11); and 16,384 (control colostrum). Elevated titers of BRV neutralizing (VN) antibodies were present in VLP colostrum (98,000), and SA11 colostrum (25,000), but not in CLP colostrum (1400), compared to colostrum from nonvaccinates (2081). Calves were orally inoculated with virulent IND BRV at 2 days of age and challenged at post-inoculation day (PID) 21. Calves were monitored daily for diarrhea and faecal BRV shedding through PID 10 and post-challenge day (PCD) 10. After colostrum feeding, the IgG1 antibody titers were highest in serum and faeces of calves fed VLP and CLP colostrum, but VN and IgA antibodies were highest in calves fed VLP colostrum. After BRV inoculation, calves fed colostrum from vaccinated cows had significantly fewer days of BRV-associated diarrhea and BRV shedding than control calves. All calves fed VLP colostrum were protected from diarrhea after BRV inoculation; two calves shed BRV. In the CLP colostrum group, one calf developed BRV-associated diarrhea and all calves shed virus. In the SA11 colostrum group, three calves developed BRV-associated diarrhea and four calves shed virus. BRV-associated diarrhea and shedding occurred in 9 of 10 control calves. Active IgM antibody responses occurred in faeces and/or serum of most calves after BRV inoculation. However, the highest active antibody responses (IgM and IgG1 in serum, and IgM, IgG1 or IgA in faeces) after BRV inoculation were in calves fed control or no colostrum, in association with clinical diarrhea in most of these calves. After challenge at PID 21, BRV-associated diarrhea and shedding were of short duration or absent, in all groups. These results demonstrate the efficacy of colostrum from VLP vaccinated cows to provide heterologous, passive protection against BRV diarrhea and shedding in calves. In comparison, calves fed CLP or SA11 colostrum were only partially protected against BRV diarrhea or shedding.

Animals↗

PubMed2000

Circulating rotavirus-specific antibody-secreting cells (ASCs) predict the presence of rotavirus-specific ASCs in the human small intestinal lamina propria.

Rotaviruses are the most important cause of infectious diarrhea in children throughout the world. Protection is most likely mediated by small-intestinal virus-specific IgA. However, neither fecal nor serum virus-specific IgA clearly correlates with protection against challenge. The capacity of rotavirus-specific antibodies and rotavirus-specific antibody-secreting cells (ASCs) in the circulation to predict the presence of ASCs in the intestines of children was evaluated. Mononuclear cells from intestinal biopsy samples and blood from 21 children were enriched for CD38, a marker of terminally differentiated B cells, and evaluated for the presence of virus-specific and total IgA- and IgG-secreting cells, by ELISPOT assay. Serum virus-specific IgA and IgG levels were determined by ELISA. The ratio of virus-specific to total IgA-secreting cells in the blood correlated with that found in the small, but not large, intestine. In contrast, serum rotavirus-specific IgA correlated less well with the presence of virus-specific ASCs in the small intestine.

ADP-ribosyl Cyclase↗