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PubMed2005

Bovine ephemeral fever in Taiwan (2001-2002).

Bovine ephemeral fever (BEF), a vector-borne disease of cattle, is caused by the Ephemerovirus of the family Rhabdoviridae. In the past 40 years, Taiwan has had seven BEF epizootics, and we have previously reported the first five. This study summarizes the 2001 and 2002 epizootics; conducted case-control serologic studies on 10 herds involved in the 2001 epizootic; determined whether the recent BEF viruses have varied significantly; and discusses the relationship between epizootic patterns and possible variant BEF viruses. For mature cows that had received at least 2 doses of vaccine before the study, a negative correlation between the prevaccinated (the 3rd dose and after) serum neutralization antibody (SNA) titers and their postvaccinated peak rates was found. When prevaccinated SNA levels were at < or = 32, their postvaccinated SNA levels increased significantly faster (P<0.01) than for those at > or = 32. The glycoprotein gene of isolates from 1999, 2001, and 2002 had a 99.2-99.9% homology, without consistent amino acid variations in the neutralization sites. Phylogenetic analysis of Taiwanese isolates revealed 2 distinct clusters, the 1983-1989 and 1996-2002 isolates. Cross-neutralization tests confirmed the glycoprotein gene sequence analysis results. In conclusion, annual boosters at SNA levels > 32, at more than 2 doses, or at intervals shorter than 6 months are not advisable. The occurrence of frequent small epizootics implies the dominance of BEF virus over host immunity, but not a variant virus.

Animals↗
PubMed1961

Studies of smallpox antibody levels of sera from samples of the vaccinated adult population of Madras.

In view of the endemicity of smallpox in Madras and the poor take rate on revaccination with the lymph in current use, it was felt that the immunity of the general adult population might be relatively low. In an attempt to obtain some measure of this, sera were obtained from over 300 adults in Madras for the estimation of serum antibody. The results of neutralization tests against variola virus made on these sera showed that about 10% of those who had good scars from infant vaccination showed little or no antibody. A comparison of the antibody titres of adult sera showing good antibody levels when used undiluted was made with that of sera from smallpox convalescents. The antibody level in the latter sera was 20-100 times that of the sera of adult persons who showed good vaccination scars.

Adult↗
PubMed1992

Antigenic change in feline calicivirus during persistent infection.

To determine if antigenic variation occurred during persistent infection of cats with feline caliciviruses (FCV), nine persistent (progeny) isolates from nine different carrier cats were compared antigenically to the original infecting parent strain, FCV 255, by two-way cross-neutralization tests with rabbit antisera. Five of the nine progeny viruses isolated 35 to 169 days after initial infection were antigenically different from the parent strain. These five isolates represented four distinct antigenic phenotypes. The emergence of four distinctly different antigenic variants from a single parent strain indicates that FCV, like many other RNA viruses, exhibits considerable antigenic heterogeneity during replication in its natural host, and supports the hypothesis that antigenic variation contributes to chronic FCV infection.

Animals↗
PubMed1985

Neutralization microtest with human coxsackievirus and echovirus serotypes.

Sets of 600 single sera from healthy individuals of various age and 458 paired sera from patients treated for diseases of various etiology were examined using a neutralization microtest technique employing prototype collection strains CA9, CB1-CB5, E1-E9, E11-E14, E17, E19, E24 and E26. Totally, 34,862 monotype neutralization tests were carried out in this study. In the set of single serum samples the lowest proportion of sera reacting with any of the enterovirus serotypes used was encountered in the group of youngest children up to the age of 2 years. This group of children showed also the highest proportion of sera free of type-specific antibody. Taken together, these sera reacted most frequently with serotypes CA9 and CB4. In the set of paired sera significant rises in antibody titre to one enterovirus serotype were recorded in 105 instances, in association with simultaneous nonsignificant rises against one or several other serotypes in 64 instances. In additional 12 paired sera there was evidenced a significant rise of antibody titre to more than one serotypes, combined in 8 of these with simultaneous nonsignificant rises to further serotypes. This neutralization microtest technique with a set of enterovirus serotypes is believed to represent a servicable diagnostic tool in determining the cause of enteroviral infection, in spite of the estimated 10% failure to establish the exact serotype responsible. The results yielded by the serologic examination of single serum samples are not considered as reflecting the actual seroconversion rates in the general population.

Adolescent↗
PubMed1980

Various immune reactions in hamsters after immunization with murine tumour cells and their roles in rejection.

Hamsters immunized with a murine tumour via the cheek pouch showed accelerated rejection following rechallenge with the same kind of tumour cells. Regional lymph node cells exhibited a suppressive effect on tumour growth in in vivo neutralization tests although cytotoxicity was not detected in such a population by 51Cr-release tests. Cytotoxicity became detectable using glass-nonadherent peritoneal exudate in cells in the 51Cr-release test, when the peritoneal cavity was used for primary immunization or rechallenge. These results suggested that lymphocytes acquired cytotoxic capacity at the site of xenograft rejection after close contact with target cells. Capacities to elicit delayed footpad reaction and to suppress tumour growth at rechallenge developed in hamsters exposed to 600 rad before immunization. Their capacity to produce cytotoxic antibody was abolished completely by such irradiation.

Animals↗
PubMed2003

Spontaneous production of anti-IFN-alpha and anti-IL-12 autoantibodies by thymoma cells from myasthenia gravis patients suggests autoimmunization in the tumor.

Myasthenia gravis (MG) is mediated by autoantibodies to the acetylcholine receptor (AChR), expressed in muscle and rare thymic myoid cells. Most early-onset cases show thymic lymph node-type infiltrates, including pre-activated plasma cells spontaneously producing anti-AChR antibodies. Since these are not evident in the associated thymomas found in another 10% of MG patients, AChR-specific B cells must be autosensitized elsewhere. Unexpectedly, at diagnosis, >70% of MG/thymoma patients also have high-titer neutralizing autoantibodies to IFN-alpha, and >50% to IL-12; moreover, titers increase strikingly if the thymomas recur, indicating a closer tumor relationship than for anti-AChR. To investigate this, we have measured autoantibody production by cells cultured from thymomas, any available thymic remnants and blood, with or without the B cell stimulant pokeweed mitogen (PWM). To check autoantibody specificity and clonal origins, we isolated Fabs from two combinatorial libraries from producer thymus/thymoma cells. Surprisingly, thymoma cells spontaneously produced antibodies to IFN-alpha and/or IL-12 in >40% of seropositive cases, showing typical plasma cell behavior, whereas they produced anti-AChR only after PWM stimulation. We isolated 15 combinatorial Fabs to IFN-alpha (versus only one to AChR). Their strong binding in radio-immunoprecipitation and Western blots implies high affinities. The four Fabs tested neutralized anti-viral actions of IFN-alpha. The diverse V genes clearly showed ongoing antigen-driven selection. These results imply pre-activation in situ by native IFN-alpha/IL-12 expressed within a 'dangerous' tumor microenvironment. With these molecules, it should be easier to identify provoking cell type(s) that may give novel additional clues to autoimmunization against T-cell epitopes from the more complex AChR.

Adult↗
PubMed2006

A pseudotype vesicular stomatitis virus containing Hantaan virus envelope glycoproteins G1 and G2 as an alternative to hantavirus vaccine in mice.

We examined whether a vesicular stomatitis virus (VSV) pseudotype bearing the hantavirus envelope glycoproteins (GPs) G1 and G2 (VSVdeltaG*HTN) could be used as a safe and effective alternative to native hantavirus. Mice were immunized with purified particles of VSVdeltaG*HTN. After the second immunization, all mice produced anti-GP antibody as detected in ELISA and a neutralization test. After the third immunization, the mice were challenged with Hantaan virus. Neither anti-NP antibody production nor Hantaan virus-specific CD8 T-cell reactions were detected in these mice. The present study demonstrated the potential of using a pseudotype VSV system as a tool for developing a hantavirus vaccine.

Animals↗
PubMed1994

Characterization of neutralization epitopes on the VP7 surface protein of serotype G11 porcine rotaviruses.

Rotavirus strain A253, isolated from the faeces of a diarrhoeic piglet in Venezuela, was classified as serotype G11 by cross-neutralization studies and by comparison of the deduced amino acid sequence of the VP7 surface protein. The epitopes involved in neutralization of the two G11 porcine rotavirus strains A253 and YM were analysed using neutralization-resistant mutants selected with seven neutralizing monoclonal antibodies (MAbs), monotype-specific (M-) MAbs and serotype-specific (S-) MAbs, produced against VP7 of strain A253. Cross-neutralization tests and sequence analysis of the escape mutants selected from strains A253 and YM indicated the presence of two antigenic sites, one common to both M-MAbs and S-MAbs in region A (positions 87, 91 and 96) and the other defined by one S-MAb in region C (position 223). All A253 variants selected with M-MAbs and two S-MAbs, although having different amino acid substitutions, had a change at amino acid position 87, whereas YM variants involved residues 91 and 96, part of the same antigenic site. Compared to strain A253, the YM stain presents an amino acid substitution at position 87 and was not recognized by M-MAbs. These results suggest that in the VP7 of G11 serotype specificity, the amino acid at position 87 is an important component of a neutralization site associated with region A and the intraserotypic variation between strains A253 and YM may account for the selection of mutations at different positions by a single MAb.

Amino Acid Sequence↗
PubMed2003

A 3-year pilot study of sentinel dairy herds for vesicular stomatitis in El Salvador.

The occurrence of vesicular stomatitis (VS) was investigated in El Salvador through monthly visits to 12 sentinel cattle operations located in four different departments. Management, environmental, and spatial data were collected. Heifers were enrolled on the operations and were examined and bled monthly for 3 years. Two competitive ELISAs were used to detect antibodies on each sample for each serotype of VS virus (VSV). On 8 of the 12 operations, small terrestrial rodents were trapped, blood samples collected, and antibodies to both VS serotypes evaluated using a serum-neutralization test for each virus serotype. Similar to other studies of VS in Central America, the seroprevalence of the New Jersey serotype was higher than the seroprevalence to the Indiana serotype. An outbreak of VS appeared to occur in the Department of Sonsonate in the summer of 1999. We confirmed that VS is endemic in the four departments investigated in El Salvador.

Animals↗
PubMed2000

Isolation of Hendra virus from pteropid bats: a natural reservoir of Hendra virus.

Since it was first described in Australia in 1994, Hendra virus (HeV) has caused two outbreaks of fatal disease in horses and humans, and an isolated fatal horse case. Our preliminary studies revealed a high prevalence of neutralizing antibodies to HeV in bats of the genus PTEROPUS:, but it was unclear whether this was due to infection with HeV or a related virus. We developed the hypothesis that HeV excretion from bats might be related to the birthing process and we targeted the reproductive tract for virus isolation. Three virus isolates were obtained from the uterine fluid and a pool of foetal lung and liver from one grey-headed flying-fox (Pteropus poliocephalus), and from the foetal lung of one black flying-fox (P. alecto). Antigenically, these isolates appeared to be closely related to HeV, returning positive results on immunofluorescent antibody staining and constant-serum varying-virus neutralization tests. Using an HeV-specific oligonucleotide primer pair, genomic sequences of the isolates were amplified. Sequencing of 200 nucleotides in the matrix gene identified that these three isolates were identical to HeV. Isolations were confirmed after RNA extracted from original material was positive for HeV RNA when screened on an HeV Taqman assay. The isolation of HeV from pteropid bats corroborates our earlier serological and epidemiological evidence that they are a natural reservoir host of the virus.

Animals↗
PubMed1977

The nature of Pseudomonas aeruginosa strain PAO bacteriophage receptors.

Receptors for phages specific to Pseudomonas aeruginosa strain PAO were studied. Phages 16, 44, 109, F8, and PBI are lipopolysaccharide (LPS) specific as shown by neutralization tests. The PhI50's of the LPS, adsorption rate constants with strain PAO and the plaque morphologies of these five phages were quite similar. Phages 1214 and 7 also appear to be LPS-specific on the basis of host-range studies. Phage 73 is pilus-specific, while phages 21 and 68 fall into a group which does not attach to pili, flagella, or LPS. A theoretical approach to the interpretation of phage-cell interactions is presented.

Bacteriophages↗
PubMed1987

A collaborative report: rhinoviruses--extension of the numbering system from 89 to 100.

To define the number of rhinovirus serotypes, cross neutralization tests and characterization studies were completed on 25 candidate prototype rhinoviruses submitted to a third phase of a collaborative program. Based on the results, 11 distinct prototype strains were designated and the numbering system was extended to include 100 rhinoviruses. In addition, recent evidence indicates that over 90% of rhinoviruses isolated in three areas of the country could be typed with antisera for rhinovirus types 1-89.

Neutralization Tests↗
PubMed1991

Trial of Edmonston-Zagreb measles vaccine in infants aged under nine months.

Due to the recent finding that most infants in developing countries have lost maternal antibody for measles before nine months of age, immunization of infants younger than the recommended age of nine months would help reducing the incidence of measles in these endemic areas. We conducted a trial of Edmonston-Zagreb measles vaccine which is the strain that may be more immunogenic in young infants than the widely used Schwarz strain. Forty-five infants with mean age of 25 weeks received a dose of Edmonston-Zagreb vaccine. Antibody levels were measured, using plaque neutralization test, before and about 3 months after vaccination at which mean age was 38 weeks. The seroconversion rate was 89%. Only two infants (4.4%) had immunity before vaccination. Fifteen infants (33.33%) reported some adverse reactions including fever (13.33%), rhinorrhea (8.89%), rash (4.44%) and local reactions (22.22%). All of the reactions resolved spontaneously. We conclude that Edmonston-Zagreb measles vaccine is efficacious and safe in infants aged under nine months.

Age Factors↗
PubMed1999

Evaluation of an enzyme-linked immunosorbent assay based on binding inhibition for type-specific quantification of poliovirus neutralization-relevant antibodies.

To detect neutralization-relevant antibodies against 3 types of poliovirus (PV) without using tissue cultures and live viruses, an enzyme-linked immunosorbent assay (ELISA) based on monoclonal antibody-binding inhibition was evaluated using sera from 80 vaccinated Japanese children and 60 Pakistani poliomyelitis patients. Compared with the neutralization test, the sensitivity of the inhibition ELISA was 100% (111/111) for detection of anti-PV1 antibody, 98.3% (118/120) for anti-PV2, and 96.5% (82/85) for anti-PV3, and the specificity was 93.1% (27/29), 100% (20/20), and 92.7% (51/55), respectively. Thus, the inhibition ELISA showed excellent potential as a seroepidemiologic tool in both vaccinated and naturally-infected populations.

Animals↗
PubMed1993

Monoclonal antibody-based blocking enzyme-linked immunosorbent assay for specific detection and titration of peste-des-petits-ruminants virus antibody in caprine and ovine sera.

A blocking enzyme-linked immunosorbent assay (B-ELISA), using two neutralizing monoclonal antibodies (MAbs), was established and compared with the virus neutralization test (VNT) for detecting specific peste-des-petits-ruminants virus (PPRV) antibody in caprine and ovine sera. This technique was developed because VNT, the only available specific serological test for PPRV and the cross-reactive rinderpest virus (RPV), is time-consuming and unaffordable for most laboratories in regions where both peste des petits ruminants and rinderpest occur. The test depends on the blocking of the binding of the MAb to a specific epitope in the presence of positive serum. Test conditions were optimized by using peste-des-petits-ruminants and rinderpest sera that were known to be VNT positive and negative. A blocking format, in which serum is preincubated with a solid-phase PPRV antigen and then incubated with the MAb, yielded levels of sensitivity and specificity superior to those of a competitive format, in which the two reagents are added simultaneously. A threshold value of 45% inhibition, representing the mean for a negative population (n = 277) plus 2.7 standard deviations, was adopted for routine screening. A total of 605 serum samples were screened by B-ELISA and the VNT. The sensitivity and specificity of B-ELISA relative to the VNT were 90.4 and 98.9%, respectively. Of 264 field serum samples tested, 11 (4.2%) could not be assayed by the VNT because of contamination or cytotoxicity; the overall agreement quotient between results of the two tests (n = 253) was 0.91. A high correlation (r>/=0.98) was observed between B-ELISA and the VNT for endpoint titration of sera (n=57). Because B-ELISA proved to be nearlyas sensitive and specific as the VNT while being simpler and more rapid, it would be an adequate substitute for the VNT for assessing herd immune status and for epidemiologic surveillance.

Animals↗
PubMed1976

Kinetics of the response of spleen cells from tumor-bearing animals in an in vivo tumor neutralization assay.

The behavior of spleen cells from tumor-bearing mice, vis-à-vis isologous tumor cells, was investigated by means of an in vivo adoptive neutralization test. C3H/eB mice were challenged with tumor cells from a chemically induced fibrosarcoma. Spleens from these animals were removed at weekly intervals following tumor inoculation, mixed with tumor cells, and tested for their influence on tumor growth in syngeneic recipient mice. Two phases in the reactivity of spleen cells from tumor-bearing mice were clearly distinguishable. In a first stage of tumor growth, these mice yielded specific tumor-inhibitory cells conferring protection. Subsequently, the protective activity declined leading to a second phase characterized by tumor enhancement. Both protective and enhancing activities were shown to be mainly dependent on the presence of T cells.

Animals↗
PubMed2006

Characterization of the sigmaB-encoding genes of muscovy duck reovirus: sigmaC-sigmaB-ELISA for antibodies against duck reovirus in ducks.

The sigmaB/sigmaC-encoding genes of muscovy duck reovirus (DRV) S12 strain were cloned, sequenced, and expressed in Escherichia coli. The sigmaC-encoding gene of DRV showed only 21-22% identity to that of avian reovirus (ARV) at both nucleotide and amino acid level. The sigmaB-encoding gene of DRV comprised 1163bp with one open reading frame (ORF). The ORF comprised 1104bp and encoded 367 amino acids with a predicted molecular mass of 40.44 kDa. A zinc-binding motif and a basic amino acid motif were found within the predicted amino acid sequence of sigmaB. The identities between the S12 and ARV were 59.3-64.0% and 60.9-62.5%, respectively, at the nucleotide and deduced amino acid levels. Phylogenetic analysis of the sigmaB-encoding gene sequence indicated that S12 separated as a distinct virus relative to other avian strains. The expressed sigmaB/sigmaC fusion proteins in E. coli could be detected, approximately 45 and 50kDa, respectively, by duck anti-reovirus polyclonal serum. In addition, an ELISA (sigmaB-sigmaC-ELISA) using the expressed sigmaB-sigmaC proteins as coating antigen for detection of antibodies to DRV in ducks was developed. In comparison with the virus neutralization test and agar gel immuno-diffusion test (AGID), the sigmaB-sigmaC-ELISA showed perfect specificity and sensitivity. The sigmaB-sigmaC-ELISA did not react with the antisera to other duck pathogens, implying that these two proteins were specific in recognition of DRV antibodies. Taken together, the results demonstrated that sigmaB-sigmaC-ELISA was a sensitive and accurate method for detecting antibodies to DRV.

Amino Acid Sequence↗
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↗