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PubMed2004

Homotypic and heterotypic serum isotype-specific antibody responses to rotavirus nonstructural protein 4 and viral protein (VP) 4, VP6, and VP7 in infants who received selected live oral rotavirus vaccines.

Homotypic and heterotypic serum isotype-specific antibody responses to rotavirus enterotoxin nonstructural protein (NSP)-4, independent neutralization antigens viral protein (VP)-4 and VP7, and group A rotavirus common antigen VP6 were analyzed by an immunocytochemistry assay in infants who received 1 of several live oral rotavirus vaccines. Significant serum immunoglobulin (Ig) A and IgG antibody responses to homotypic and/or heterotypic NSP4s of genotype [A], [B], or [C] were detected after vaccination. The magnitude of antibody responses to homotypic and heterotypic NSP4s was not significantly different, irrespective of the NSP4 genotype of the administered vaccine strain. In addition, there were no significant differences between IgA antibody responses to homotypic and heterotypic VP7s. In contrast, IgA antibody responses to VP4 were predominantly homotypic. IgA antibody responses to VP7 were lower in magnitude than those to VP4 but were comparable to those to NSP4. Antibody titers to homotypic and/or heterotypic NSP4s were positively correlated with those to VP6 before and after vaccination.

Antibodies, Viral↗

PubMed1983

Protection studies in colostrum-deprived piglets of a bovine rotavirus vaccine candidate using human rotavirus strains for challenge.

Studies were performed to evaluate the potential use of the bovine RIT 4237 rotavirus strain as a vaccine candidate against infection with human rotaviruses. Initial experiments revealed that colostrum-deprived piglets were susceptible to infection with several human strains, except for those belonging to subgroup 1. Subsequently, different immunization procedures with RIT 4237 were studied in this animal model. It was found that a two-dose administration, either given intramuscularly (twice) or once intramuscularly and once intragastrically, was necessary to induce a significant serum antibody response. Finally, the protective effect of the latter vaccination schedules against subgroup 2 and 3 rotavirus strains of human origin was evaluated by artificial challenge. In both cases, prior administration of live RIT 4237 significantly decreased fecal shedding of the challenge virus when compared with control animals.

Animals↗

PubMed1984

Prevention of rotavirus-induced diarrhea in neonatal mice born to dams immunized with empty capsids of simian rotavirus SA-11.

Female CF-1 mice were immunized subcutaneously with a rotavirus vaccine consisting of noninfectious, purified empty capsids from simian rotavirus SA-11 and were given a booster dose at 14-17 days of gestation. The vaccinated animals developed high titers of rotavirus-specific IgG in colostrum and milk. Virus-specific IgA titers were significantly (150-fold) lower, and virus-specific IgM was not detected. These results contrast with those obtained in natural and experimental infections of the gastrointestinal tract with epizootic-diarrhea infant-mouse ( EDIM ) virus; in such cases, intestinal and lacteal virus-specific antibody is primarily of the IgA isotype. Suckling mice born to vaccinated dams were challenged with infectious EDIM virus and monitored for the development of diarrheal disease; these neonates acquired maternal antibody and did not develop diarrhea.

Animals↗

PubMed1988

Seroconversion patterns to four human rotavirus serotypes in hospitalized infants with acute rotavirus gastroenteritis.

We studied rotavirus-specific antibodies in paired sera from 71 hospitalized infants with acute rotavirus gastroenteritis. Most of the infants were less than six months old. Infants with serological evidence of a secondary rotavirus infection were excluded. With an enzyme-linked immunosorbent assay, 46% of the 71 infants studied showed specific IgM in convalescent sera. Titers of specific IgG and IgA increased in 7% and 2% of the infants, respectively. The presence of specific IgM correlated positively with age and severity of clinical symptoms. With a neutralization test, 59% of the infants showed a seroconversion: 20% to a single serotype (7% to serotype 1, 7% to serotype 3, and 6% to serotype 4), 21% seroconverted to two serotypes (nearly exclusively to serotypes 1 and 3), and 18% seroconverted to three serotypes (exclusively to serotypes 1, 3, and 4). No infant seroconverted to serotype 2 or to the heterologous (bovine) serotype 6.

Age Factors↗

PubMed1990

Rotavirus-specific antibody response in saliva of infants with rotavirus diarrhea.

The reliability of saliva as an indicator of rotavirus infection was assessed among 15 infants (3-12 months) with rotaviral and 15 with nonrotaviral diarrhea. Paired salivary samples collected during acute and convalescent phases were tested for rotavirus-specific IgA and IgM by an ELISA. The sensitivity of IgA or IgM alone to predict infection was 53.3% and 46.6%, respectively; used in conjunction, the sensitivity rose to 80%. It seems that infants with rotaviral diarrhea mount mucosal antibody responses as reflected in their saliva; possibly salivary antibodies could be used to evaluate vaccine "take" in rotavirus vaccine trials.

Antibodies, Viral↗

PubMed1990

[Comparison of clinical features of rotavirus infection with rotavirus antigen titers in feces obtained during the acute phase].

When fecal specimens obtained from infants with acute gastroenteritis during the acute phase between January 1986 and March 1987 were examined by enzyme immunoassay (EIA), 72 cases were found to have the rotavirus antigen. Such 3 clinical signs as diarrhea (D), fever (F) and vomiting (V) were all observed in 38 cases. These cases showing DFV syndrome demonstrated the highest geometric mean titer of rotavirus antigen among the rotavirus antigen-positive cases. Geometric mean titers (GMTs) were calculated to be as high as more than 10(3.1) in 26 of the 38 DFV cases (68%) and in 11 of 34 cases with DV, DF, FV or D syndromes (32%), showing a significant difference between the 2 groups (p less than 0.005). GTMs were slightly higher in cases having fever of 38 degrees C or more, than in those having fever of less than 38 degrees C.

Acute Disease↗

PubMed1998

Development of a fluorescent focus identification assay using serotype-specific monoclonal antibodies for detection and quantitation of rotaviruses in a tetravalent rotavirus vaccine.

A fluorescent focus identification assay (FFIDA) was developed for use in experimental studies and for quantitation of the components in a tetravalent live oral rotavirus vaccine. The assay utilizes four serotype-specific neutralizing monoclonal antibodies (MAb) to detect and quantify individual rotaviruses by immunofluorescence staining of fixed virus-infected monkey kidney cells. In mixed virus infections, all four MAb, W1 (serotype 1), 1C10 (serotype 2), R1 (serotype 3), and S4 (serotype 4), specifically stain the relevant homologous serotype without exhibiting any cross-reactivity against the other serotypes. Furthermore, the test is sensitive enough to differentiate at least twofold (0.3 log) differences in virus titer. The results of testing four individual experimental vaccine lots three or more consecutive times showed that all four lots contained similar proportions of the four vaccine strains as detected by the classical plaque neutralization identification test. The rapidity and efficiency of the FFIDA are desirable attributes that make it suitable for use in studies requiring identification and quantitation of one or more of the four major rotavirus serotypes.

Animals↗

PubMed1985

Porcine rotavirus-like virus (group B rotavirus): characterization and pathogenicity for gnotobiotic pigs.

A rotavirus-like virus (RVLV) was isolated from a diarrheic pig from an Ohio swine herd. This virus infected villous enterocytes throughout the small intestine of gnotobiotic pigs and induced an acute, transitory diarrhea. Complete virions were rarely observed in the intestinal contents of infected animals; the predominant particle detected by immune electron microscopy was a corelike particle 52 nm in diameter. The genome of the porcine RVLV was composed of 11 discrete segments of double-stranded RNA that produced an electropherotype distinct from the genome electropherotypes of reovirus, rotavirus, and porcine pararotavirus. Porcine RVLV was antigenically unrelated to rotavirus, porcine pararotavirus, or reovirus but was antigenically related to a bovine RVLV.

Animals↗

PubMed1992

Hemagglutination by a human rotavirus isolate as evidence for transmission of animal rotaviruses to humans.

Human rotavirus strain Ro1845, which was isolated in 1985 from an Israeli child with diarrhea, has a hemagglutinin that is capable of agglutinating erythrocytes from guinea pigs, sheep, chickens, and humans (group O). Hemagglutination was inhibited after incubation with hyperimmune sera or in the presence of glycophorin, the erythrocyte receptor for animal rotaviruses. These results suggest that Ro1845 is an animal rotavirus that infected a human child.

Animals↗

PubMed1997

Isolation from diarrheal and asymptomatic kittens of three rotavirus strains that belong to the AU-1 genogroup of human rotaviruses.

A survey of 143 stool specimens collected during a 17-month period in Japan from diarrheal and asymptomatic kittens identified three rotavirus strains that were similar in their genomic RNA constellation to human rotavirus AU-1 (G3P3[9]), lending strong support to the view that rotaviruses belonging to the same genogroup are circulating in both the human and the feline population.

Animals↗

PubMed2003

Changing patterns of rotavirus genotypes in ghana: emergence of human rotavirus G9 as a major cause of diarrhea in children.

Genotyping of human rotaviruses was performed on 312 rotavirus-positive samples collected from 2,205 young children with diarrhea in the Upper East District of Ghana, a rural community. Of the 271 (86.9%) rotavirus strains that could be VP7 (G) or VP4 (P) characterized, 73 (26.9%) were of G9 specificity. The predominant G9 genotype was G9P[8], which constituted 79.5% of all G9 strains detected, followed by G9P[6] (12.3%), G9P[10] (2.7%), and G9P[4] (1.3%). G9 strains with mixed P types constituted 2.7% of all G9 strains found in the study. All the G9P[8] strains had a long RNA electrophoretic pattern with VP6 subgroup II specificity. Four G9 isolates, GH1319, GH1416, GH3550, and GH3574, which were selected based on the abundance of stool material and were representative of the three electropherotypes observed, were cloned and sequenced. The Ghanaian isolates shared more than 98% sequence nucleotide homology with other G9 strains from the United States (US1205), Malawi (MW69), Brazil (R160), Japan (95H115), and Nigeria (Bulumkutu). However, they showed only 95% nucleotide homology with the Thai G9 strain Mc345. Phylogenetic analysis of the nucleic acid sequence revealed the existence of at least three clusters, with Ghanaian strains forming one cluster, Nigerian and Brazilian strains forming a second cluster, and U.S., Malawian, and Japanese strains forming a third.

Antigens, Viral↗

PubMed1990

Cell culture propagation of calf rotavirus and detection of rotavirus specific antibody in colostrum and milk of cows and buffaloes.

A bovine rotavirus was adapted to serial propagation in Madin Darby bovine kidney (MDBK) cells. Virus of cell culture origin when examined by electron microscopy exhibited characteristic rotavirus morphology and had a typical RNA electropherotype on polyacrylamide gel electrophoresis. Mean IgG concentration in the first colostral whey was 62.80 +/- 6.56 mg/ml in cows and 104.40 +/- 31.12 mg/ml and 2.52 +/- 0.28 mg/ml respectively. The geometric mean neutralising antibody titre against bovine rotavirus in the first colostral whey was 279 in cows and 139 in buffaloes. It declined to 10 and 13 (respectively) by the tenth day.

Animals↗

PubMed1993

[Present findings on rotaviruses with emphasis on rotaviruses in swine].

A summary of thirty pages, concerning rotaviruses, i.e. RNA-viruses, the most frequent causal agents of infectious diarrhoea in children and young farm animals. Most of the chapters are generally focused on rotavirus morphology, structure and biology. Chapter No. 6 is especially detailed, trying to offer a complete summary of available diagnostic methods. The final chapters concern in greater detail the problems of rotaviruses in pigs, such as the immunity, prevention and treatment.

Animals↗

PubMed1993

Gene 5 of the IDIR agent (group B rotavirus) encodes a protein equivalent to NS34 of group A rotavirus.

Gene 5 of the IDIR strain of group B rotavirus was cloned, sequenced, and expressed in RBC lysates in order to identify the coding assignment. The complete nucleic acid sequence of IDIR gene 5 included 1306 bases and encoded a single long open reading frame of 390 amino acids with a predicted molecular weight of 44.8 kDa. Comparison of the predicted amino acid sequence indicated substantial identity with the NS34 protein of group A rotaviruses (GAR). Like GAR NS34, the product deduced from IDIR agent gene 5 was predicted to exhibit a high degree of alpha helix secondary structure and a relatively low pl (4.6). In vitro translation of gene 5 resulted in synthesis of a protein which was specifically immunoprecipitated by convalescent antibody obtained from infant rats following infection with the IDIR agent, confirming that this product was of viral origin. Under non-reducing conditions, products as large as 200 kDa were also noted in SDS-PAGE. Formation of oligomers by GAR NS34 has previously been observed, indicating that this feature is probably a general characteristic among rotavirus NS34 and may relate to the functional role of the protein in viral replication.

Amino Acid Sequence↗

PubMed1987

Bovine rotavirus with rearranged genome reassorts with human rotavirus. Brief report.

Bovine rotavirus (brv) possessing a rearranged genome [Hundley et al. (1985), Virology 143: 88-103] was found to reassort with tissue culture-adapted group A human rotavirus carrying a standard genome. The rearranged part of the brv genome containing segment 5-specific sequences was exchanged with the normal RNA segment 5 of the human rotavirus in some of the reassortants.

Animals↗

PubMed1994

Molecular determinants of rotavirus virulence: localization of a potential virulence site in a murine rotavirus VP4.

The molecular basis of pathogenesis in vivo for a virulent mouse rotavirus (MRV) and a less virulent bovine rotavirus (BRV) were compared under in vitro and in vivo conditions. Obvious differences in the mobility of several genomic RNA segments were observed in one-dimensional gels. Under in vitro conditions, partial proteolytic peptide mapping identified differences between the two outer capsid proteins of these virus and no difference in inner capsid protein was observed. Since it has been observed by us and others that the gene coding for VP4 protein plays a significant role in determining virulence, the variability observed in the present study between the 84 k proteins (VP4) provided a basis for further investigations in order to locate a potential virulence determinant. A comparison of the carboxypeptidase digests of the MRV- and BRV-VP4 revealed an area of variability between amino acids 307 and 407, which may represent a site of virulence determinant. Under in vivo conditions the virulence of both parenteral BRV and MRV isolates and their corresponding reassortants (with replaced gene 4) were studied in murine and bovine hosts. Like their parents, BRV and MRV isolates, reassortants obtained by replacement of gene 4 in BRV with MRV gene 4 indicated that the dose of the virus isolate used and the clinical outcome in vivo was determined by gene segment 4. The implications of these findings to elucidate the molecular basis of pathogenesis of rotaviruses are discussed.

Animals↗

PubMed1985

Virus enzyme-linked cell immunoassay (VELCIA): detection and titration of rotavirus antigen and demonstration of rotavirus neutralizing and total antibodies.

Virus enzyme-linked cell immunoassay (VELCIA) for detection and titration of rotavirus antigen has been developed. Wild-type porcine rotavirus antigen can be detected and titrated directly from the fecal material within 24 h. Porcine OSU strain can be titrated higher than 10(-8). The method has also been introduced for the demonstration of rotavirus neutralizing and total antibodies. In VELCIA the advantages of the cell culture system for virus isolation are combined with enzyme immunodetection and spectrophotometrical reading of the test.

Antibodies, Viral↗

PubMed2005

Molecular characterization of the rotavirus NSP4 enterotoxin homologue from group B rotavirus.

The RNA segment (Gene 10) from a human group B rotavirus which encodes the homologue of the rotavirus enterotoxin (NSP4) has been cloned and sequenced. The gene is of the same length (751 nucleotides) as its better-characterized group A rotavirus counterpart but shows minimal homology (approximately 10%) to it at the primary sequence level. Despite this low level of sequence homology, secondary structure predictions for the group B protein (ADRV-NSP4) showed a close similarity of structural features with the group A protein. Full-length ADRV-NSP4 was expressed in Escherichia coli with an amino terminal 6xHis tag that was used to purify it to homogeneity. The cytotoxicity of the purified protein was examined in a rapid dye-uptake assay that assesses membrane permeability and was found to be comparable to its group A counterpart.

Amino Acid Sequence↗