Detection of bovine rhinotracheitis virus antibody by neutralizing test and ELISA in experimentally infected rabbits.

The antibody response of rabbit to infectious bovine rhinotracheitis (IBR) virus was examined. Two rabbits were inoculated with IBR virus strain Los Angeles into the trachea and intravenously, and intravenously two times, respectively. The patterns of antibody titers in the rabbits measured by neutralization test and ELISA were similar to those in the case of bovine. The antibody was detected after the inoculation, and much more antibody was detected after the second inoculation. The fact suggests that rabbits are a very useful laboratory animal for IBR virus infection studies.     

Antibody response to glycoprotein E after bovine herpesvirus type 1 infection in passively immunised, glycoprotein E-negative calves

This study was conducted to determine whether young calves with maternal antibodies against bovine herpesvirus type 1 (BHV-1) but without antibodies against glycoprotein E (gE) can produce an active antibody response to gE after a BHV-1 infection. Five calves received at birth colostrum from gE-seronegative cows which had been vaccinated two or three times with an inactivated BHV-1, gE-deleted marker vaccine. After inoculation with a wild-type virulent strain of BHV-1, all the passively immunised gE-negative calves shed virus in large amounts in their nasal secretions. All the calves seroconverted to gE within two to four weeks after inoculation and then had high levels of gE antibodies for at least four months. The development of an active cell-mediated immune response was also detected by in vitro BHV-1-specific interferon-gamma assays. All the calves were latently infected, because one of them re-excreted the virus spontaneously and the other four did so after being treated with dexamethasone. The results showed that under the conditions of this work the gE-negative marker could also distinguish between passively immunised and latently infected calves.     

Protective immunity following immunisation of pigs with aerosol of Actinobacillus pleuropneumoniae serotype 2

To introduce antigen to the respiratory mucosa, killed Actinobacillus pleuropneumoniae with quil A as adjuvant was administered to pigs as an aerosol. Immunisation by this aerosol induced a marked IgA response in the bronchoalveolar and nasal fluids, and in the serum. Following challenge with live bacteria two weeks after the last exposure to the aerosol, the immunised pigs were protected from the severe pleuropneumonia which developed in non-immunised pigs. The immunised pigs had lower antibody titres in the mucosal fluids and serum after exposure to the challenge. The immune response after experimental infection of non-immunised animals was a weak IgA antibody response in the bronchoalveolar and nasal fluids, whereas the systemic immune response after challenge included both IgA and IgG antibodies.     

Potential of DNA-mediated vaccination for equine herpesvirus 1.

The potential of DNA-mediated immunisation to protect against equine herpesvirus 1 (EHV-1) disease was assessed in a murine model of EHV-1 respiratory infection. Intramuscular injection with DNA encoding the EHV-1 envelope glycoprotein D (gD) in a mammalian expression vector induced a specific antibody response detectable by two weeks and maintained through 23 weeks post injection. Immune responses were proportional to the dose of DNA and a second injection markedly enhanced the antibody response. EHV-1 gD DNA-injected mice developed neutralising antibodies, and a predominance of IgG2a antibodies after the DNA injection was consistent with the generation of a type 1 helper T-cell (Th1) response. Following intranasal challenge with EHV-1, mice immunised with 50 microg of EHV-1 gD DNA were able to clear virus more rapidly from lung tissue and showed reduced lung pathology in comparison with control mice. The data indicate that DNA-mediated immunisation may be a useful strategy for vaccination against EHV-1.     

Effect of infections with swine fever virus on immune functions. III. Antibody response to lipopolysaccharide and sheep red blood cells

In order to determine the effect of infections with low-virulent swine fever virus (SFV) on antibody responses, pigs were administered lipopolysaccharide (LPS) or sheep red blood cells (SRBC), 2 days after infection. Infected pigs showed an enhanced primary response to LPS late during infection. The secondary response to LPS seemed to be unaffected. Both the primary and secondary antibody response to SRBC appeared to be enhanced rather than depressed in infected pigs. These in vivo findings suggest that pigs infected with low-virulent SFV do not develop a depression of B lymphocyte function.     

Humoral immune response in goats immunised with cathepsin L1, peroxiredoxin and Sm14 antigen and experimentally challenged with Fasciola hepatica

The humoral immune response was analysed in goats immunised with FhCL1, FhPrx, Sm14, and experimentally challenged with Fasciola hepatica. All immunised animals developed significant levels of anti-fluke specific antibodies and those immunised with FhCL1 showed the highest antibody titre. After experimental infection, an increase in the antibody level was detected only in goats immunised with FhCL1. In the adjuvant-control animals, the experimental challenge induced significant production of specific antibodies against FhCL1, FhPrx and Sm14. While liver fluke specific humoral responses were seen in all groups, no significant protection in any of the vaccinated groups was found.     

Comparison of three adjuvants used to produce polyclonal antibodies to veterinary drugs

Two commercially available adjuvants, Gerbu LQ 3000 and Montanide ISA 50V, were assessed as potential replacements for Freund's adjuvant by evaluating their efficacy in the production of polyclonal antibodies to veterinary drugs in rabbits. The aim was to find an adjuvant that could produce a similar (or enhanced) immune response in the host animal without the undesirable side effects associated with Freund's complete and incomplete adjuvant. The assessment involved the examination of each injection site and the characterisation of the resultant antibodies with regards to antibody titre and sensitivity. It was found that the rabbits immunised with Gerbu adjuvant produced some of the most sensitive antibodies. However, titres were relatively low and adverse effects at injection sites were relatively common. Montanide adjuvant produced no adverse effects and the related antibodies were found to be of adequate sensitivity when compared to those from rabbits immunised with Freund's. It was concluded that Montanide ISA 50V could be considered as a suitable replacement to Freund's for the production of polyclonal antibodies, to low molecular weight compounds in rabbits.     

猪伪狂犬病不同佐剂灭活疫苗对兔免疫原性初探

采用分离鉴定的猪伪狂犬病毒（HB—J株）以4种佐剂研制成4批灭活疫苗,以研究其对兔的免疫原性。疫苗分别免疫PRV抗体阴性兔后,用乳胶凝集试验法（LAT）和中和试验法（SNT）对试验兔进行血清抗体检测;于免疫后21、28d对试验兔分别进行攻毒,观察兔保护情况。试验结果表明,兔对不同佐剂的猪伪狂犬病疫苗均产生良好的免疫应答反应,且当兔免疫后血清凝集价≥1：32或血清抗体中和指数≥1479或中和价≥1：16时,可以抵抗10LD50剂量强毒的攻击;当兔免疫后血清凝集价≥1：64或血清抗体中和指数≥2187或中和价≥1：32时,可以抵抗100LD50剂量强毒的攻击;4种佐剂灭活疫苗均具有良好的免疫效果。     

Canine parvovirus: interaction between passive immunity and virulent challenge

Two groups of puppies, one passively immunised by the administration of hyperimmune serum and the other with natural maternally derived antibody, were inoculated orally with virulent canine parvovirus of faecal origin. Serum antibody titres declined more rapidly in both groups after challenge than before. The dogs became clinically affected but the onset of clinical signs, seroconversion and faecal excretion of virus was delayed when compared to controls. It is postulated that this rapid decline of antibody was due to its sequestration by virus after the initial phase of viral replication in the lymphoid tissues. These findings have important implications. The incubation period of the disease is prolonged, making it more difficult to estimate accurately the time of infection in clinically affected animals. Furthermore, the more rapid decline of maternally derived antibody, which could occur in endemically infected premises, may complicate immunisation programmes based on the isolation and segregation of puppies in anticipation of a predicted decline in maternally derived antibody before vaccination.     

Humoral and cellular factors affecting the neutrophil response of the locally immunised mammary gland to staphylococcal infection

Locally immunising the non-lactating ovine mammary gland by infusing killed Staphylococcus aureus enhances neutrophil accumulation in mammary secretion during subsequent staphylococcal infection. Immunological factors influencing this increased neutrophil response were studied in the present experiments. Glands locally immunised with killed Brucella abortus supported a greater neutrophil response to staphylococcal infection than did glands immunised with killed S. aureus. An enhanced neutrophil response to staphylococcal infection was recorded in 3 of 7 ewes locally immunised with zymosan. Passive immunisation by intramammary infusion of secretions from immunised glands conferred an enhanced neutrophil response during staphylococcal infection. Absence of haemolytic complement in secretions of immunised glands suggested complement was not implicated in the response. Secretions of immunised glands contained elevated concentrations of mononuclear cells. When exudates rich in mononuclear cells were established by infusion of endotoxin into non-immunised glands there was no increase in the neutrophil response to subsequent staphylococcal infection. However, when the mononuclear cell concentration was elevated by intramammary infusion of staphylococcal antigens in systemically immunised ewes there was an increase in the neutrophil response to subsequent infection. Thus humoral and cellular characteristics of the locally immunised mammary gland influence the kinetics of the neutrophil influx during staphylococcal infection.     

Effect of corticosterone infusion on plasma corticosterone concentration, antibody production, circulating leukocytes and growth in chicken lines selected for humoral immune responsiveness

The effect of corticosterone on antibody production was studied in chicken lines selected for humoral immune response. 2. Twelve cockerels (33 days old) from lines selected for high or low antibody responses after immunisation with sheep red blood cells (SRBC) were implanted with mini-infusion pumps delivering corticosterone or vehicle continuously for 14 d. 3. Three days after implantation, the chickens were immunised intramuscularly with 0.25 ml packed SRBC. Blood samples were taken before implantation, before immunisation and 3, 5, 7 and 11 d after immunization. 4. Corticosterone infusion induced higher plasma corticosterone concentrations and heterophil/lymphocyte ratios than infusion of vehicle only. Growth was considerably depressed and relative weights of the thymus, bursa of Fabricius and spleen were less in the corticosterone-infused chickens. 5. An effect of corticosterone on antibody production could not be demonstrated, and differences between selection lines were unaffected.     

Comparison of the agar gel immunodiffusion test and ELISA in the detection of bovine leukosis virus antibody in cattle persistently infected with bovine virus diarrhoea virus

When six cattle persistently infected with bovine virus diarrhoea virus (BVDV) were inoculated with lymphocytes infected with bovine leukosis virus (BLV), a depressed antibody response to BLV was observed by ELISA which was due to a decrease in IgG1 synthesis. The ELISA was more sensitive and more reliable than the agar gel immunodiffusion (AGID) test in detecting BLV infection in cattle persistently infected with BVDV. Decreased antibody responses were manifested in the AGID test by negative, inconclusive or weakly positive reactions: only two of the six cattle developed antibodies that generated positive AGID reactions.     

Comparison of the immunogenicity of Newcastle disease virus strains V4, Hitchner Bl and La Sota in chickens

The immunogenicity of three Newcastle disease virus (NDV) strains--V4, Hitchner B1 and La Sota, was assessed in chickens that had varying levels of maternal antibody to the virus. Chickens were immunised at different ages by the eye drop and aerosol methods. The immune response of chickens to similar doses of the strains was affected by the presence of maternal antibody. Both the level of maternal antibody and the strain of virus used affected the result. Strain La Sota was least affected as it took higher levels of maternal antibody to depress response to it than were required to have a similar effect on Hitchner B1 and V4. Strain V4 was the strain most affected by circulating maternal antibody. The results demonstrated that strain V4 was less immunogenic than Hitchner B1 and La Sota when used in chickens with maternal antibody to NDV.     

The Efficacy of a Bivalent Vaccine Against Pasteurellosis and Rabbit Haemorrhagic Disease Virus

Rabbit haemorrhagic disease virus (RHDV) and Pasteurella multocida bacteria cause severe losses among rabbit populations. The efficacy of a recently developed bivalent vaccine against pasteurellosis and RHDV was investigated. Doses exceeding 2 haemagglutinating units (HU) of viral antigen were sufficient to protect rabbits against infection with RHDV. The bivalent vaccine appeared to be safe for use in all age groups of rabbits, including pregnant females, even after treatment with 20 times the normal vaccine dose. Rabbits injected with 8 or 4 HU of bivalent vaccine showed high antibody titres against both organisms for 9 months after inoculation. The antibody levels against RHDV in young rabbits at 30 days of age were elevated when they originated from mothers with high antibody titres. The most suitable period for vaccination of offspring appeared to be around 50 days of age. The bivalent vaccine against pasteurellosis and RHDV combined speed and longevity of the immune response. Immune protection against pasteurellosis and RHDV can thus be achieved with only one manipulation.     

Serum containing ovine IgG2 antibody specific for maedi visna virus envelope glycoprotein mediates antibody dependent cellular cytotoxicity

Antibody-dependent cell-mediated cytotoxicity (ADCC) specific for maedi visna virus (MVV) has never been described. The IgG antibody response to MVV is restricted to an IgG1 response whilst MVV specific IgG2 is never seen in persistently infected sheep. To determine whether the isotypic restriction of the antibody response is responsible for the lack of ADCC, an ADCC assay was developed using polyclonal serum raised to recombinant MVV ENV protein. Sheep immunised with a recombinant GST:SUenv fusion protein in complete Freund's adjuvant produced an antibody response which contained IgG1 and IgG2 antibodies. The activity of this serum in an ADCC assay was compared to serum from persistently infected sheep. Serum from immunised sheep mediated ADCC reactions whilst no activity was ever seen in persistently infected sheep serum. IgG2 may therefore be the possible effector isotype for ADCC reactions against MVV. Failure of the IgG2 dependent ADCC system in vivo may contribute to the persistence of MVV-infected macrophages in vivo.     

Clinical and immunologic responses of calves with colostrally acquired maternal antibody against parainfluenza-3 virus to homologous viral infection.

Exposure of colostrum-deprived calves and calves with colostrally acquired maternal antibody to aerosols of parainfluenza-3 (PI-3) virus resulted in signs of infection, leukopenia, and shedding of virus from the nasal passages. However, infection was not as severe in calves with colostrally acquired maternal antibody as it was in colostrum-deprived calves which did not have antibody to PI-3 virus before they were exposed. All calves responded immunologically to PI-3 virus, as indicated by resistance to challenge exposure and subsequent development of virus-neutralizing antibody. However, levels of serum and nasal secretion (NS) antibody at 30 days after viral exposure were lower in calves with colostrally acquired maternal antibody than in colostrum-deprived calves, and a serum antibody response in the former was primarily indicated by an anamnestic response after challenge exposure. After calves were challenge exposed to PI-3 virus, serum and NS antibodies were increased in all calves, but antibody titers were generally lower for calves that had colostrally acquired maternal antibody before their exposure than for those that acquired antibody only after PI-3 viral infection.     

Immunoglobulins and anti-Marek's disease virus antibody synthesis in chickens after passive immunization with immunoglobulin Y anti-Marek's disease virus antibody.

The effect of passive immunization with immunoglobulin Y (IgY) antibody against Marek's disease virus (MDV) was examined in MDV-susceptible chickens. The production of IgY, immunoglobulin M, and probably also immunoglobulin A was depressed in passively immunized chickens when compared with that in MDV-exposed chickens which had not been given IgY anti-MDV antibody. In passively immunized chickens, the synthesis of immunoglobulin M and IgY anti-MDV antibodies in response to MDV infection also was delayed as determined by agar gel precipitin and indirect fluorescence antibody tests.     

Studies on the methods of preparation of rinderpest hyperimmune sera in rabbits

The procedures for the preparation of the rinderpest hyperimmune sera in rabbits were studied by comparing the sera from rabbits immunised by three different schedules of inoculations. The best sera for use in immunodiffusion tests were obtained from rabbits inoculated first with rinderpest hyperimmune serum and lapinised virus, and then with lapinised virus mixed with oil adjuvant twice at weekly intervals. Those rabbits which received additional one or two intravenous inoculations with lapinised virus yielded satisfactory sera for use in the diagnosis of rinderpest by immunodiffusion technique.     

Duration of excretion of virulent Newcastle disease virus following challenge of chickens with different titres of serum antibody to the virus

Virulent Newcastle disease virus (NDV) was isolated from susceptible and immune chickens following intra-ocular challenge with the Essex '70 strain. Challenge virus was isolated from the trachea and cloaca of susceptible birds until they died 7 to 9 days after challenge. This virus was isolated from immunised chickens for up to 14 days after challenge. The duration of excretion was influenced by the prechallenge serum antibody titre to NDV. It persisted longest in chickens with titres of 2(3) to 2(7) and decreased in length and frequency from chickens with titres in the range 2(8) to 2(12). Chickens with pre-challenge titres of 2(3) to 2(5) developed 2- to 3- fold increases in post-challenge titres, whereas those with higher pre-challenge titres had smaller proportional increases in titre. Excretion of virulent virus from immunised birds should be considered in the development of Newcastle disease control programs.     

Effect of passively-acquired antibodies and vaccination on the immune response to contagious ecthyma virus

Lambs which received colostrum from ewes vaccinated with contagious ecthyma (CE) virus and other lambs vaccinated with CE virus were compared for serum anti-CE immunoglobulin (Ig)G levels, delayed-type hypersensitivity (DTH) responses to CE viral antigen, and protective immunity to challenge with CE virus. Ewes vaccinated 3-4 weeks prior to parturition transferred CE antibody to lambs via colostrum. Although these lambs had higher levels of antibody at challenge than lambs vaccinated when 1-4 days old, only the vaccinated lambs were protected against challenge with CE virus at 1 month of age. Furthermore, the presence of colostrum-derived maternal antibody prevented an active antibody response in lambs to vaccination and/or challenge with CE virus, except where pre-inoculation titres were low. In contrast, the DTH response to CE viral antigen and induction of protective immunity by CE vaccination were not impaired by passively-acquired antibody. Actively immunised lambs could be distinguished from those only receiving passively-acquired antibody by the DTH response to heat-killed CE viral antigen.