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Antibody response to 146S particle, 12S protein subunit and isolated VP1 polypeptide of foot-and-mouth disease virus type Asia-1.

The antibody response to foot-and-mouth disease virus (FMDV) antigens of type Asia-1 in guinea-pigs was studied by micro-serum neutralization test (MSNT) and enzyme-linked immunosorbent assay (ELISA). One inoculation of as little as 1 microgram of binary ethyleneimine (BEI)-inactivated 146S virus particles in guinea-pigs elicited enough neutralizing antibodies to protect them against challenge with virulent virus. However, one inoculation of live 146S virus particles elicited higher levels of neutralizing antibodies in guinea-pigs than that of inactivated 146S particles. One inoculation of 12S protein subunits in guinea-pigs elicited only non-neutralizing anti-12S antibodies detected by ELISA. Similarly, non-neutralizing anti-VP1 antibodies were detected by ELISA after one inoculation of VP1. However, multiple inoculations of 12S or VP1 elicited measurable neutralizing antibodies in guinea-pigs that protected them against challenge.

Animals↗

Evaluation of vaccine Encepur Adult for induction of human neutralizing antibodies against recent Far Eastern subtype strains of tick-borne encephalitis virus.

We studied humoral immune response of 44 volunteers from Primorsky krai (Russia) immunized with the vaccine Encepur Adult. Induction of the humoral response towards the recently isolated tick-borne encephalitis virus (TBEV) strains P-69, P-202, and P-73 was evaluated by neutralization test and enzyme immunoassay. These strains belong to Far Eastern TBEV subtype based on their genotype and antigenic structure but maintain significant genetic and antigenic variability. The average geometric titers of neutralizing antibodies to P-69, P-202, and P-73 strains were 1:28, 1:34, and 1:128, respectively. The percentage of volunteers with neutralizing antibodies to these strains after complete course immunization was 63.9, 97.6, and 95.5%, respectively. We concluded that Encepur Adult vaccine induced pronounced humoral immune response towards genetically and antigenically heterogeneous strains of the Far Eastern TBEV subtype.

Adult↗

Isolation of a virus closely related to gibbon ape leukaemia virus from cells infected with virus (HL-23V) released by human leukaemic cells.

Canine thymus cells infected with virus (HL-23V) produced by human acute myelogenous leukaemia cells in culture were shown in previous reports to produce transforming and non-transforming type C virus similar or identical to the simian sarcoma virus complex SSV(SSAV) and to induce tumours in marmoset monkeys (Bergholz et al. 1977a). In these earlier studies the appearance of breakthrough foci at low dilutions of antiserum in neutralization tests with high-titred anti-SSV(SSAV) serum suggested the presence of another virus, distinct from SSV-(SSAV). We now report the isolation of this component and, by comparative neutralization analysis, demonstrate that it is most closely related to gibbon ape leukaemia virus (GALV). It is distinguished from SSV(SSAV) by kinetics of neutralization and molecular hybridization experiments. This component was readily cloned both from virus produced by HL-23V chronically-infected canine thymus cells established by Teich et al. (1975) when HL-23V was first isolated and from virus produced by HL-23V-induced marmoset tumour cells in culture. The presence of this component in the original leukaemic cell cultures is discussed.

Animals↗

Biological properties and viral surface antigens of Burkitt lymphoma- and mononucleosis- derived strains of Epstein-Barr virus released from transformed marmoset cells.

Three strains of Epstein-Barr virus (EBV), two from Burkitt lymphoma (BL) and one from infectious mononucleosis (IM) were used to transform separate cultures of the same batch of primary marmoset leukocytes, and the viruses released from the transformants were compared. The three viruses shared properties of the transforming biotype of EBV, namely, stimulation of DNA synthesis and immortalization of cord blood leukocytes, and failure to induce "early antigen" in lymphoblast lines. All viruses produced more virus in transformed marmoset cells than in transformed human cells, as measured by the number of EBV genomes detected by complementary RNA/DNA hybridization, by virus capsid antigen expression, or by released virions and biologically active virus. Reference human sera and sera from primary EBV infections were used to compare the three virus strains in a virus neutralization test based on inhibition of stimulation of DNA synthesis. Specimens taken late in convalescence from patients with mononucleosis and sera from marmosets experimentally infected with virus from a patient with mononucleosis neutralized the homologous virus, as well as the two virus strains isolated from patients with BL. This finding indicates that viral antigens that elicit neutralizing antibodies are shared among the strains. However, in certain sera the neutralizing-antibody titer against one strain was consistently higher than against another strain. Furthermore, sera taken early after onset of IM contained low levels of neutralizing antibody against IM-derived virus, but failed to neutralize BL-derived virus. These latter findings suggest the existence of heterogeneity among surface antigens of EBVs. The results emphasize the biological and antigenic similarity of EBV isolates from BL and IM and do not suggest major subtype variations. It remains to be determined whether antigenic diversity such as described or virus genome variation detectable by other means is epidemiologically significant.

Animals↗

[The nature of nonspecific reactions in designing erythrocyte diagnostic agents for determining the HBs antigen].

The nature of nonspecific reactions observed in reverse passive hemagglutination test was studied. Nonspecific reactions were found to be associated with the use of different ingredients in construction of immunodiagnostic preparations for HBs antigen and with the presence in the examined sera of heterologous hemagglutinins, rheumatoid factor, and bacterial contamination. Therefore, positive results of hemagglutination should be verified for specificity by neutralization tests with HBs antigen.

Animals↗

Antigenic differences between strains of Newcastle disease virus.

Comparison of strains of Newcastle disease virus by kinetic neutralization tests and immunodiffusion tests (after virion disruption by ether and Tween 80) enabled antigenic differences to be demonstrated between four out of five strains tested. No correlation was demonstrated between the antigenic structure and virulence of these strains.

Antigens, Viral↗

A modified procedure for the propagation of wild type Epstein-Barr virus in cultures of marmoset blood cells.

Productive infection of marmoset blood mononuclear cells by the Epstein-Barr virus was generally achieved by the co-cultivation method. By introducing 2 modifications, the success rate of infection was increased from 6% to 79%. The modifications consisted of the selection of human lymphocyte donors to serve as carriers of Epstein-Barr virus in cocultures, and the addition of cyclosporin A to culture media. So far, 10 of 10 wild type oropharyngeal EBV have been propagated successfully in cultures of marmoset blood mononuclear cells by the modified procedure. Fragment length polymorphism study failed to reveal any difference between viral genomes in human lymphocytes and that in marmoset blood mononuclear cells. Antigenic analysis of 8 wild strains showed that all were related to the B95-8 strain of Epstein-Barr virus by the neutralization test.

Animals↗

Dengue tetravalent DNA vaccine inducing neutralizing antibody and anamnestic responses to four serotypes in mice.

We developed a dengue tetravalent DNA vaccine consisting of plasmids expressing premembrane and envelope genes of each of four serotypes of dengue viruses. BALB/c mice immunized twice with the tetravalent vaccine at a dose of 100 microg (25 microg for each serotype) using a needle-free jet injector developed neutralizing antibodies against all serotypes. There was no interference among the four components included in this combination vaccine. Tetravalent vaccine-immunized mice showed anamnestic neutralizing antibody responses following challenge with each dengue serotype: responses to challenges from serotypes different to those used for neutralization tests were also induced.

Animals↗

Neutralization epitope diversity of coxsackievirus B4 isolates detected by monoclonal antibodies.

Only recently have we begun to fully realize the diversity of virions within a single human isolate of the group B Coxsackieviruses. These intratypic (strain) differences may be one of the important factors influencing pathogenesis, e.g., myocarditis vs. pancreatitis. Yet, until the virion strains within a type can be well differentiated, a thorough analysis of the pathogenic potential of each is not possible. We compared two human isolates, JVB and Edwards, and 15 isolates derived from these two in search of determinants of strain specificity and to evaluate the diversity/stability of neutralization epitopes on the virions comprising each isolate. Polyclonal antisera failed to show strain specific determinants. However, monoclonal antibodies directed to individual neutralization epitopes defined strain specific differences. Plaque reduction neutralization tests with these monoclonal antibodies allowed the quantitation of the expression of epitopes on the virions in each isolate. These data helped to establish the limits of diversity and stability of the neutralization epitopes of Coxsackievirus B4 virions and describe the changes in epitopes among laboratory isolates.

Antibodies, Monoclonal↗

Apparent lack of neutralizing antibodies in Aleutian disease is due to masking of antigenic sites by phospholipids.

It is generally accepted that Aleutian disease virus (ADV) cannot be neutralized by antibodies either in vivo or in vitro. We found several ways to demonstrate neutralization of ADV by specific antibodies from mink. It was essential to make ADV monodisperse by treatment with sodium lauroyl sarkosyl or n-butanol or by filtration through 0.05-micron membranes before neutralization tests. In kinetic experiments, there was a 95% loss of virus infectivity within the first 5 min of reaction, but a resistant fraction of about 1% remained after 1.5 hr of incubation. Neutralization titers between 1:160 and 1:640 were found in sera from naturally and experimentally infected mink. A positive relation was consistently found between neutralization and ELISA titers. Furthermore, separation of phospholipids from ADV was shown by thin-layer chromatography of butanol-extracted virions. By reconstitution of monodispersed ADV with various lipids, phospholipids were found to interfere with virus neutralization by attachment to the virus surface.

Aleutian Mink Disease↗

Serologic response to canine distemper viral antigens in gnotobiotic dogs infected with canine distemper virus.

The humoral immune response to canine distemper viral antigens in gnotobiotic dogs experimentally infected with canine distemper virus was studied by the complement fixation and serum neutralization tests. Antibody titers measured by both serologic methods varied inversely with the severity of disease produced. Recovered dogs demonstrated the highest titers of antibody, whereas fatally infected dogs had little or no antibody activity in their sera. A third group of dogs, characterized by chronic persistent infection, had intermediate levels of anitbody to canine distemper virus. Preliminary characterization of the viral antigens involved in the complement fixation test indicated that at least two antigenic components were involved. One antigen was soluble in ether and was heat-labile, whereas the other was relatively heat-stable and was unaffected by treatment with ether. No evidence for serologic cross-reaction between viral and central nervous system components was found. A vigorous antibody response to envelope antigen determinants in addition to core determinants distinguished immune dogs from persistently infected dogs. The results of this study suggest that the inability to produce antibodies to envelope antigens may be a crucial factor in the establishment of a persistent infection with canine distemper virus in these dogs.

Animals↗

Molecular identification of new picornaviruses and characterization of a proposed enterovirus 73 serotype.

Enteroviruses (EV) have traditionally been identified by using serotype-specific antisera in a virus-neutralization test. Three EV strains isolated in California, USA, in 1955, 1964 and 1978, and a 1995 Oman isolate, were found to be antigenically related to one another; however, the strains were not neutralized by standard EV typing antisera, suggesting that they may represent a new EV serotype. The isolates were characterized genetically by RT-PCR coupled with amplicon sequencing and comparison to a database of enterovirus nucleotide sequences. The strains were 75.3 to 87.2% identical to one another in complete VP1 nucleotide sequence, but no more than 68% identical in sequence to the prototype strain of any EV serotype. Their complete capsid sequences were closely related to one another, but only distantly related to those of any EV prototype strain. The California and Oman isolates were most closely related to members of EV cluster B, suggesting that they are unclassified members (i.e. a new serotype) of cluster B. The complete genome sequence was determined for one isolate, CA55-1988, and the predicted polyprotein sequence was 86.5 to 89.2% identical to those of other cluster B EV and 56.7 to 61.9% identical to the polyprotein sequences of EV belonging to other clusters. Isolation of this new EV serotype from samples obtained on two continents and over a period of 40 years suggests continued circulation over a wide geographical area. In keeping with standard picornavirus nomenclature, we propose that this new serotype be named 'enterovirus 73' (EV73).

Antigens, Viral↗

Naturally occurring murine leukemia viruses in wild mice: characterization of a new "amphotropic" class.

A new class of murine leukemia viruses, isolated from wild Mus musculus trapped in California, is described. These viruses, designated "amphotropic," replicate in mouse, rabbit, mink, human, guinea pig, and rat cells, but not in hamster, quail, or duck cells. They show N-tropism for mouse cells, and do not trigger the XC cell response. They are distinct by interference and virus neutralization testing from the previously recognized mouse-tropic and xenotropic MuLV classes. Mouse-tropic viruses occuring along with three of the four amphotropic isolates were found to be distinguishable by virus neutralization from other mouse-tropic murine leukemia virus strains of laboratory mouse origin.

Animals↗

Molecular and antigenic characterization of porcine rotavirus YM, a possible new rotavirus serotype.

In 1983, we isolated a porcine rotavirus (strain YM) that was prevalent in several regions of Mexico, as judged by the frequency of its characteristic electropherotype. By a focus reduction neutralization test, rotavirus YM was clearly distinguished from prototype rotavirus strains belonging to serotypes 1 (Wa), 2 (S2), 3 (SA11), 4 (ST3), 5 (OSU), and 6 (NCDV). Minor, one-way cross-neutralization (1 to 5%) was observed when antisera to the various rotavirus strains were incubated with rotavirus YM. In addition, the YM virus was not neutralized by neutralizing monoclonal antibodies with specificity to serotypes 1, 2, 3, and 5. The subgroup of the virus was determined to be I by enzyme-linked immunosorbent assay. To characterize the serotype-specific glycoprotein of the virus at the molecular level, we cloned and sequenced the gene coding for VP7. Comparison of the deduced amino acid sequence with reported homologous sequences from human and animal rotavirus strains belonging to six different serotypes further supported the distinct immunological identity of the YM VP7 protein.

Amino Acid Sequence↗

False positive antibody results against human T-cell lymphotropic virus in patients with severe acute respiratory syndrome.

Taiwan suffered from the outbreak of severe acute respiratory syndrome (SARS) in 2003. Our laboratory performed a series of virology and serology tests for SARS patients admitted to our hospital. Cross-reactivity was found when testing for antibody against human T-cell lymphotropic virus (HTLV) in one patient with SARS. Therefore, antibodies against HTLV were examined in paired-sera from 26 SARS patients. ELISA and a neutralization test were used to measure anti-SARS antibodies. Seroconversion for antibody against SARS-CoV was observed in all patients. Surprisingly, with the use of ELISA for HTLV, sera for 13 patients were positive for HTLV (50%), and seroconversion for HTLV was also observed in 10 patients (38.5%). Western blot for HTLV on those 26 paired-sera from 13 HTLV-positive patients displayed 5 positive results for HTLV-I, 7 positive results for HTLV-II, 1 positive result for both HTLV-I and II, 9 negative results for either HTLV-I or HTLV-II, and 4 "indeterminate" results. The findings that antibody to HTLV can be detected in blood samples collected from SARS patients provide important information for safe handling of blood products. Without such knowledge, blood products can be discarded mistakenly even though they contain anti-SARS-CoV antibodies that may be potentially valuable for SARS therapy.

Adolescent↗

Genetic and antigenic analysis of betanodaviruses isolated from aquatic organisms in Taiwan.

Viral nervous necrosis (VNN) is a worldwide disease among marine fishes. In Taiwan, NNN disease was first identified in 2 species of hatchery-reared grouper, Epinephelus fuscogutatus and E. akaaya in 1994. Since then, increasing mortalities have occurred among groupers Epinephelus spp., and also among European eels Anguilla anguilla L., yellow-wax pompano Trachinotus falcatus, firespot snapper Lutaanus erythropterus B., barramundi Lates calcarifer, cobias Rachycentron canadum, humpback groupers Cromileptes altivelis and Chinese catfish Parasilurus asotus. In the present study, samples were collected from affected fishes and processed for reverse transcriptase (RT) PCR amplification and virus isolation in cell culture. Infected cells (GF-1 cell line) exhibited cytopathic-effect characteristics of grouper nervous necrosis virus (GNNV). A RT-PCR product of approximately 830 bp was amplified from the brain homogenate of tested samples and sequenced. The nucleotide and deduced amino acid sequences of the amplified RT-PCR products from all isolates were strongly homologous (> 97 %) with the corresponding region of the published sequence of red-spotted grouper nervous necrosis virus (RGNVV). Therefore, all Taiwan NNV (nervous necrosis virus) isolates studied in this report belong to the RGNNV genotype. We used 5 neutralizing monoclonal antibodies (MAbs) against GNNV to analyze the antigenic relationship of Taiwan NNV isolates and striped jack nervous necrosis virus (SJNNV). The results of neutralization tests revealed that all Taiwan NNV isolates were closely related, but antigenically different from SJNNV in 3 neutralizing epitopes. To our knowledge, this is the first description of NNV infection in European eels, yellow-wax pompano, firespot snapper, cobia and Chinese catfish, and the first reported instance of natural NNV infection in freshwater fishes causing high mortality.

Amino Acid Sequence↗

Antibody determination in an ongoing hepatitis B vaccination program.

In an interim analysis of our ongoing immunization program against hepatitis B (HB), started in early 1982, we tested 283 serum samples from 77 female and 110 male vaccinees for antibody to HB surface antigen (anti-HBs). We compared two methods, Anti-HBs EIA (ROCHE) (method 1), which is a neutralization test, and AUSABR EIA (method 2), which is a double-antigen-sandwich test. The nonresponder rate (after the 12-month booster dose of Hevac B Pasteur) was 4% in females with both methods, in males 15% measured with method 1 and 11% measured with method 2. Five healthy HBsAg carriers were detected only by method 1. When the samples were grouped according to their anti-HBs titers, method 1 measured higher in samples taken after three vaccine doses and method 2 did so in samples collected after the 12-month booster dose. This tendency was confirmed with samples from slow responders who received a 4th vaccine dose soon after the initial three doses. We therefore confirm the efficacy of the plasma-derived HBsAg vaccine and validate the assay systems used to measure anti-HBs, one parameter of immunity to HB virus infection.

Adult↗