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1.
In an attempt to produce a DNA vaccine to prevent Aujeszky's disease, the induction of immune responses against Aujeszky's disease virus (ADV) gD was investigated in mice. The plasmid was constructed by placing ADV gD gene downstream of murine cytomegalovirus immediate early promoter of expression vector pMYK, which was injected twice on the skin of mice by using a gene-gun. All mice showed neutralizing antibodies against ADV gD at 4 weeks after immunization. The induction of cytotoxic T lymphocytes and splenic natural killer cells was also observed at 6 weeks post immunization. These results indicate that ADV gD gene in the form of DNA vaccine may induce specific as well as non-specific immune responses in vivo.  相似文献   

2.
Aujeszky's disease virus (ADV) envelope glycoprotein gVI (gp50) was purified from virus-infected Vero cells by ion-exchange and immunoaffinity chromatography and its usefulness as a subunit vaccine was evaluated in active and passive immunization studies. Four-week-old piglets were immunized intramuscularly (IM) with purified gVI twice two weeks apart and challenged intranasally (IN) 10 days after the second immunization with 30 LD50 (10(8)PFU) of a virulent strain of ADV. Pigs, vaccinated with 100 micrograms of purified gVI, produced virus neutralizing antibodies and did not develop clinical signs after challenge exposure. The challenge virus was not isolated from nasal swabs and tonsils of gVI-vaccinated pigs, whereas non-vaccinated control pigs developed illness after challenge exposure with the same virulent ADV strain which was later recovered from their nasal swabs and tonsils. Pregnant sows vaccinated twice with purified gVI (IM) at a three week interval produced virus neutralizing antibodies in colostrum. Four-day-old sucking piglets born of vaccinated sows were passively protected by colostral antibodies against intranasal challenge with a lethal dose of virulent ADV. Sera from gVI-vaccinated pigs were distinguished from experimentally infected swine sera by their differential reactivity in enzyme-linked immunosorbent assay (ELISA) using four major viral glycoproteins (excluding gVI) as antigen purified by the use of lentil-lectin.  相似文献   

3.
The temperature-sensitive (ts), thymidine kinase-deficient (TK-) mutant designated ZHtsTK- strain, of Aujeszky's disease virus (ADV) was isolated from a virulent strain with the treatments using 5-bromodeoxyuridine and arabinosylthymine. The ZHtsTK- strain was easily distinguished from the other virulent ADV strains by plaque size on HmLu-1 and chicken embryo fibroblast cells and by restriction endonuclease analyses using Bam HI, Sal I and Kpn I. The ZHtsTK- strain was avirulent for mice, guinea pigs and rabbits, and produced neutralizing antibodies to ADV in these animals. The rabbits inoculated with the ZHtsTK- strain did not shed detectable amounts of virus after dexamethasone treatment. The ZHtsTK- strain was also avirulent for 5-day-old piglets and did not cause disease. No virus was detected from the piglets inoculated intramuscularly in the nasal swabs or the tissues examined on postinoculation day 9. These findings presented here suggested that there is a significant correlation between pathogenicity and properties such as ts and TK-, and the combination of ts and TK- properties plays a much larger role in reducing virulence for animals.  相似文献   

4.
Macrophage activity, cytokines serum concentration, serum neutralizing antibodies and lethality by rabies were evaluated in swiss mice experimentally infected with street rabies virus and submitted or not to antirabies vaccination and immunomodulation with P. acnes. Animals were killed at different times and serum was collected in order to evaluate cytokines concentration; peritonial and splenic macrophages were collected for macrophage activity evaluation. Greater survival rates higher IL-10 and low IL-6 serum concentration were observed in vaccinated animals treated using P. acnes.  相似文献   

5.
Subclinical low-prevalent Aujeszky's disease (AD) serological test reactors test reactors are defined as those few swine within a qualified AD virus (ADV)-negative herd that have antibodies to wild type virus. However, clinical signs of the associated diease are not observed in these putatively infected swine or elsewhere in the herd. Twelve such animals, including 7 previously vaccinated with a genetically modified ADV, were identified in Illinois (USA) during a 2.5 year period. The humoral immune responses of the 5 nonvaccinated swine were assessed by an enhanced virus neutralization test and a radioimmunoassay. Anti-ADV antibodies were determined to be present in the serum from 4 of these swine. Attempts to isolate ADV by in vitro and in vivo inculations of cell cultures and weanling mice, respectively, of tonsillar and trigeminal nerve ganglionic tissue preparations from each animal (vaccinated and nonvaccinated) were unsuccessful. Tonsillar and trigeminal nerve ganglionic tissues of each animal were screened for the presecence of wild type and/or vaccine viral genomes by a soluble polymerase chain reaction (PCR) coupled with Southern hybridization. Unique PCR primers were used distinguish between wild type and vaccine viral DNAs. Additional PCR procedure, which amplifiers a portion of the essential and highly conserved viral gp50 gene, also was employed in an effort to detect viral genomes. Wild type viral DNA was found in the tissues from at least 5 of the vaccinated and 3 of the nonvaccinated swine. These results indicate that such animals should be considered as being infected with ADV. Further, these findings emphasize the need to develope highly specific and sensitive antemortem testing methods for accurate assessment of ADV infection in herds containing such subclinical, yet serologically positive, swine.  相似文献   

6.
Groups of striped skunks (Mephitis mephitis) were inoculated intramuscularly with graded doses of street rabies virus. At various intervals after inoculation, saliva and sera were tested for rabies virus and neutralizing antibodies, respectively. Skunks that developed rabies were killed in terminal stages of the disease and the following examinations were made: titers of virus and antibody in submandibular salivary glands and brain, extent of immunofluorescence in submandibular salivary glands, and histologic examination of various tissues.

Skunks that received inocula containing 4 × 104 to 4 × 105 mouse intracerebral lethal dose50 (MICLD50) had detectable serum neutralizing antibodies by 7–12 days postinoculation; however, most of the skunks that received lower doses (40 to 4 × 103 MICLD50) did not have detectable serum neutralizing antibodies until clinical signs began. In the salivary glands, slight and extensive immunofluorescence corresponded to high and low titers of tissue neutralizing antibody. Also low viral titers were associated with high tissue neutralizing antibody titers. There was a close correlation between viral titers in right and left submandibular salivary glands.

The results suggest that the immune response can impede the process of infection of the salivary glands resulting in lack of antigen or low amounts of antigen in this tissue. This could occur through interference with centrifugal neural transport of virus and/or neutralization of virus during transfer from neural elements to epithelial cells. Lack of infectious virus or low viral titers in salivary glands containing antigen and high levels of tissue neutralizing antibodies can be caused partly by postmortem virus neutralization (during viral titration).  相似文献   


7.
Eight 2-month-old merino lambs were inoculated intranasally with different (10(2.0)-10(5.0)TCID50) amounts of Aujeszky's disease virus (ADV). Electron microscopic studies indicated that ADV replicated in extra-neural sites, in the epithelial cells of the mucosa of the upper and lower respiratory tract. Although the virus was excreted continuously in nasal discharges, horizontal transmission to contact lambs failed. The surviving exposed and contact lambs had no demonstrable antibodies against ADV and they were susceptible when challenged by ADV. However, the virus was transmitted to susceptible pigs in contact with the exposed lambs. One of the five contact pigs showed characteristic clinical signs of Aujeszky's disease, developed a nonsuppurative meningoencephalomyelitis and ADV was recovered from the brain, nasal discharge and other organs. Restriction enzyme analysis of DNA from this virus confirmed the sheep origin of the isolate. The other 4 pigs seroconverted. ADV infection in sheep is therefore a possible source of infection for pigs, but the lack of horizontal transmission in sheep was confirmed.  相似文献   

8.
The release of tumor necrosis factor-alpha (TNF-) from cultured bovine alveolar macrophages (BAM) was evaluated following stimulation of BAM with bovine herpesvirus-1 (BHV-1), parainfluenza-3 (PI-3) virus, bovine respiratory syncytial virus (BRSV), Escherichia coli 0111:B4 endotoxin, Pasteurella haemolytica type 1 endotoxin, Pasteurella multocida endotoxin, and virus/endotoxin combinations. A cytotoxic assay system using Georgia bovine kidney cells as targets was used to measure TNF- activity. The cytotoxic activity was neutralized by an anti-human TNF- monoclonal antibody.

Stimulation of BAM with 1 median tissue culture infectious dose (TCID50) of live or ultraviolet (UV)-inactivated PI-3 virus/cell resulted in release of TNF- in significantly (P<0.05) higher amounts than sham-induced BAM. The quantities of TNF- released after live or UV-inactivated BHV-1 or BRSV induction were not significantly higher than sham-induced BAM. E. coli 0111:B4, P. haemolytica type 1 and P. multocida endotoxins stimulated TNF- release in a dose-dependent manner. Sequential exposure of BAM to 1 TCID50 per cell of either live BHV-1, PI-3 virus or BRSV and then 5 μg ml−1 of either E. coli 0111:B4, P. haemolytica type 1 or P. multocida endotoxin caused a significant (P<0.05) reduction in detectable TNF- in seven of nine virus/endotoxin combinations tested, when compared with 5 μg ml−1 of endotoxin alone. Parainfluenza-3 virus/endotoxin combinations stimulated higher TNF- release when compared with other virus/endotoxin combinations. Five out of six test animals had serum-neutralizing antibodies to PI-3 virus, one out of six had serum-neutralizing antibodies to BHV-1, and two out of six had serum-neutralizing antibodies to BRSV, suggesting a possible relationship between serum neutralizing antibodies and TNF- release from in vitro cultivated BAM.  相似文献   


9.
The K strain of Aujeszky's disease virus (ADV) grown in Vero cells was used to vaccinate pigs. Following intramuscular inoculation, the pigs remained healthy, no vaccine virus was excreted and virus could be detected only at the inoculation site. One inoculation gave good protection against challenge with a virulent strain of ADV, and the amount of virulent ADV excreted was geatly curtailed. Following vaccination only low leads of serum neutralizing antibody were detected (geometric mean titre 1/2), but three weeks after challenge very high levels were found (GMT 1/1773). Intranasal vaccination gave similar results. There was minimal excretion of vaccine virus. The clinical reaction on challenge was less severe than in the intramuscularly challenged group, although lower antibody levels were detected three wekks following challenge (GMT 1/483). A field trial, using this strain given subcutaneously, indicated that one inoculation of this vaccine is effective.  相似文献   

10.
Several Aujeszky's disease virus (ADV) vaccination protocols of sows were evaluated with regard to the passive protection conferred on piglets in a recently built commercial farm. Three different groups of sows were vaccinated using a Bartha K-61 strain. One group received an inactivated vaccine during pregnancy and the other two groups received attenuated vaccines, either during pregnancy (day 65) or on the seventh day of lactation. At farrowing, sows vaccinated during lactation had lower seroneutralization titres than those vaccinated during pregnancy either with inactivated or attenuated vaccines. Accordingly, their piglets were the ones with lower levels of maternally transferred neutralizing antibodies. At 4 weeks of age, five piglets born of each group of sows were challenged intranasally with a neurotropic strain of ADV. Piglets born of sows vaccinated during pregnancy with inactivated and attenuated vaccines gained 1.50 kg bodyweight and 2.50 kg bodyweight during 7 days, respectively, and did not show clinical signs, while piglets from sows vaccinated during the previous lactation lost 0.60 kg and presented moderate to severe clinical signs of ADV. Vaccination of sows during pregnancy provided more protection against ADV for piglets than sow vaccination before mating. Piglets born from sows vaccinated with attenuated or inactivated vaccines did not present remarkable differences on protection.  相似文献   

11.
A liquid-phase enzyme-linked immunosorbent assay (ELISA) was developed for the detection of vesicular stomatitis virus (VSV) types New Jersey (VSV-NJ) and Indiana subtype Indiana I (VSV-IND1) antibodies in the sera of naturally and experimentally infected cattle, horses and swine. Four different VSV preparations were compared for use as antigen in the ELISA: virus used in neutralization tests, complement-fixation antigen, immunodiffusion ager gel antigen and viral glycoprotein. Comparative antibody titration results of virus neutralization (VN) and ELISA showed no statistically significant difference between serum titers obtained with the four antigens to VSV-NJ (P=0.21) and VSV-IND1 (P=0.14). The viral glycoprotein antigen was incorporated in the ELISA system because it is non-infectious and induces neutralizing antibodies. The reliability and variability of the ELISA was determined by testing 516 bovine, equine and swine sera which originated from areas free of vesicular stomatitis, and by testing 186 sera from areas where outbreaks occur. ELISA and VN results were correlated (P < 0.001 for both serotypes), and no statiscafically difference was found between replications of the tests. The ELISA allows the testing of a larger number of sera in a shorter time than is possible with the VN test and it can be used in diagnostic laboratories in VSV-free areas for the support of epidemiological surveillance programs.  相似文献   

12.
Six-Aleutian (aa)-genotype violet mink were infected intraperitoneally with the Aleutian Disease Virus (ADV) bone marrow derived isolate ADV SL3. All animals developed virus-specific antibodies and hypergammaglobulinaemia. Mortality during the fourteen week duration of the infection was 50%. The virus induced (histo)pathological lesions typical for Aleutian Disease. By immunohistochemical examination using a virus capsid-specific monoclonal antibody viral antigen was detected in lymph nodes, spleen, kidneys and once in hepatic Kupffer cells. By Southern blot and in situ hybridization studies with strand-specific RNA probes able to distinguish viral replicative forms from merely sequestered genomic DNA, ADV replication was detected in mesenteric lymph nodes and spleen. In one mink DNA replicative forms were also found in bone marrow cells or mononuclear cells of the peripheral blood, respectively. Only single-stranded viral DNA was detected in liver, kidney, gut and lung of infected animals. From Southern blot hybridization results a different, possibly organ-specific permissiveness of ADV in vivo is suggested.  相似文献   

13.
赵静  褚颖  罗均  郭霄峰 《中国畜牧兽医》2022,49(12):4776-4785
【目的】构建表达犬瘟热病毒（CDV）血凝素蛋白H基因的重组狂犬病病毒（RABV）,并研究其生物学特性及免疫原性。【方法】在RABV HEP-dG株基因组G与L基因之间插入CDV H基因,构建重组全长cDNA质粒pHEP-dG （H）。将pHEP-dG （H）与辅助质粒共同转染BHK细胞,拯救携带CDV H基因的重组RABV HEP-dG （H）。将重组病毒接种BHK细胞,分析病毒的扩散能力;将重组病毒接种小鼠,分析病毒的致病性和免疫原性。【结果】RT-PCR及直接免疫荧光显示,传至第3代的病毒仍能检测到H基因,表明CDV H基因已成功插入RABV基因组中,并在HEP-dG （H）中稳定遗传和正确表达。HEP-dG （H）在BHK细胞中的生长曲线与HEP-dG毒株相似,病毒滴度在96 h达到峰值,但HEP-dG （H）的滴度在每个时间点略低于HEP-dG。以感染复数（MOI）为0.005感染BHK细胞,HEP-dG （H）的扩散能力比HEP-dG毒株低。HEP-dG （H）与HEP-dG对6周龄成年小鼠均不致死,但HEP-dG （H）对成年小鼠体重的影响弱于HEP-dG。HEP-dG （H）与HEP-dG均能诱导小鼠产生抗RABV的中和抗体,免疫后7 d抗体已达到保护水平（0.5 EU/mL）;此外,HEP-dG （H）可诱导产生CDV中和抗体。HEP-dG （H）与HEP-dG免疫小鼠3周后,均能抵御RABV标准攻毒毒株CVS-24的攻击。【结论】本研究成功构建重组RABV HEP-dG （H）,其具有良好的免疫原性和安全性,可作为RABV-CDV新型二联基因工程候选疫苗。  相似文献   

14.
A double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed for measuring Aujeszky's disease virus (ADV) antigen concentration and an inhibition technique based on the former was developed for detection of antibodies to ADV. The results were checked by determining the cytopathic and serum neutralization titres. The correlation was satisfactory in both cases, with correlation coefficients above 0.8. When measuring ADV antigen concentration, the lower limit of detection was 10(3) TCID 50/0.2 ml. The sensitivity of ELISA in detecting antibodies to ADV was found to be superior to that of the serum neutralization test and, thus, enabled the testing of rabbit and guinea-pig sera.  相似文献   

15.
A total of 2,722 sera collected between 1963 and 1983, from 43 different species of wildlife in 11 African countries was examined for neutralising antibodies against the wildebeest-derived strain of malignant catarrhal fever (MCF) virus. Antibodies were demonstrated in 10 species of Bovidae which included eight species from the sub-family Hippotraginae and one species each from Bovinae and Antilopinae. Neutralising antibodies were also recorded in hippopotamus. It is suggested that the high prevalence of antibodies recorded in sera from waterbuck and reedbuck indicate infection with MCF. However, titres in other species may be due to antigenically related viruses.  相似文献   

16.
A total of 198 pigs with tachypnoea and temperature >/= 40 degrees C were selected on a Spanish finishing unit, and their sera were examined for antibodies to Actinobacillus pleuropneumoniae (App), porcine reproductive and respiratory syndrome virus (PRRSV), Aujeszky' disease virus (ADV), and swine influenza virus (SIV). Eighty-nine point nine per cent of the pigs were seropositive to App, 88.6 per cent to PRRS, 73.0 per cent to ADV, and 30.6 per cent to SIV. Thirty-one pigs (15.6 per cent) were seropositive for App, PRRSV, ADV and SIV, and only one (0.5 per cent) was seronegative for all. Statistical association was assessed for dual infections but it was not found in any case (P > 0.05). Other parameters (dyspnoea, nasal discharge and coughing) were also recorded, and no significant associations between them and the presence of antibodies against any of the four infections was found.  相似文献   

17.
Mechanisms contributing to the virus persistence in Aleutian disease   总被引：1，自引：0，他引：1  
In this review published results and further studies concerning the persistence of Aleutian disease virus (ADV) isolate SL3 are presented. By Southern blot and in situ hybridization with strand-specific RNA probes focal replication of ADV-DNA was demonstrated in spleen, mesenteric lymph nodes, sporadically in mononuclear cells of the peripheral blood and bone marrow cells. These findings further support the concept of the lymphotropism of ADV. All cell culture-adapted ADV strains appear to have a ts-defect. Our in vitro studies indicate that the ADV isolate G(orham) induced the synthesis of comparable amounts of viral replicative DNA and viral proteins VP1 and VP2 at the non-permissive temperature of 37 degrees C. However, the viral progeny DNA synthesis was about threefold less at 37 degrees C compared to the permissive temperature of 32 degrees C. These findings suggest that the reduced level of viral progeny DNA at 37 degrees C accounts for the reduced production of infectious ADV. Finally, we provided experimental evidence that the apparent lack of neutralizing antibodies in AD is due to the masking of critical viral epitopes by cellular phospholipids.  相似文献   

18.
ABSTRACT: Foot-and-mouth disease virus (FMDV) is a highly infectious member of the Picornaviridae inducing an acute disease of cloven-hoofed species. Vaccine-induced immune protection correlates with the presence of high levels of neutralizing antibodies but also opsonising antibodies have been proposed as an important mechanism of the immune response contributing to virus clearance by macrophages and leading to the production of type-I interferon (IFN) by plasmacytoid dendritic cells (pDC). The present study demonstrates that the opsonising antibody titres mediating enhanced IFN-α responses in pDC were similar to neutralizing titres, when antigenically related viruses from the same serotype were employed. However, sera cross-reacted also with non-neutralized isolates of multiple serotypes, when tested in this assay. Both uncomplexed virus and immune complexed virus stimulated pDC via Toll-like receptor 7. An additional finding of potential importance for strain-specific differences in virulence and/or immunogenicity was that pDC activation by FMDV strongly differed between viral isolates. Altogether, our results indicate that opsonising antibodies can have a broader reactivity than neutralizing antibodies and may contribute to antiviral responses induced against antigenically distant viruses.  相似文献   

19.
The three glycoproteins each of feline herpesvirus type 1 (FHV-1) and canine herpesvirus (CHV) were purified by affinity chromatography using glycoprotein-specific monoclonal antibodies and used individually or in combination in immunizing mice to determine their relative immunogenicity. All the glycoproteins induced detectable virus neutralizing antibodies to the homologous virus but FHV-1 gp143/108 and its cross-reacting counterpart, CHV gp145/112, elicited the highest titers not only to the homologous virus but to the heterologous virus as well. The production of ELISA antibodies after glycoprotein immunization was variable, while hemagglutination-inhibiting antibodies were produced by only 1 out of 10 FHV-1 gp60-inoculated mice. In general, the antibody titers induced by CHV glycoproteins were lower than those by FHV-1 glycoproteins. These results indicate that these glycoproteins may be useful as subunit vaccines against FHV-1 and CHV infections.  相似文献   

20.
赤羽病毒单克隆抗体的研制及鉴定   总被引：2，自引：2，他引：0  
用纯化的赤羽病毒（akabane virus,AKAV）免疫Balb/c小鼠,取小鼠脾细胞和骨髓瘤细胞SP2/0融合,经间接ELISA筛选和3次有限稀释法克隆,得到2株能稳定分泌抗赤羽病毒单克隆抗体（McAb）的杂交瘤细胞株,分别命名为AKAV McAb 3A株和2C株。ELISA试验和中和试验结果表明,本研究制备的2株McAb均具有良好的特异性,为AKAV阳性,杂交瘤细胞培养上清液抗体的效价分别为1∶640和1∶320,腹水的效价分别为1∶256000和1∶128000,亲和常数（Ka）分别为1.16×10-9和6.31×10-8 mol/L,3A株的相对亲和力大于2 C株,具有病毒中和活性,中和效价分别为1∶64和1∶32,其IgG亚类为IgG1,轻链的亚型均为kappa型,2株细胞冻存3次复苏后仍能稳定分泌抗体,表明AKAV McAb制备成功,为赤羽病快速诊断方法的研究奠定了基础。  相似文献   

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