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1.
To study the antibody response to glycoprotein I (gI) of pseudorabies virus (PRV) in maternally immune pigs, 3 groups of 6 pigs were given low doses of the mildly virulent Sterksel strain of PRV at 3 and 11 weeks of age. Group A consisted of seronegative pigs; groups B and C consisted of pigs with maternal antibodies deficient of antibodies to gI. At 3 weeks of age, 3 pigs of each group were inoculated intranasally with 10(2.5) plaque-forming units (groups A and B), or with 10(3.5) plaque-forming units (group C) of PRV. The 3 other pigs in each group were contact-exposed to the inoculated pigs. In group A, 4 of 6 pigs shed virus and all developed antibodies to gI of PRV and produced PRV-specific IgM and virus-neutralizing antibodies. In groups B and C, 10 pigs shed virus and all developed low and inconsistent titers of gI antibodies, whereas only 3 pigs produced PRV-IgM antibodies with low titers. Thus, after PRV infection of pigs with high concentrations of maternal antibodies deficient of gI antibodies, the antibody responses to PRV were severely inhibited. The pigs were reinoculated with 10(3) plaque-forming units of the same virus 8 weeks after the first inoculation. The pigs in group A did not respond at all, as they were immune. The pigs in groups B and C shed considerable amounts of virus. Three pigs had a clear secondary antibody response to gI, whereas the others developed an early to normal antibody response to gI. None of the pigs mounted a secondary neutralizing antibody response to PRV.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

2.
A collection of 90 field isolates of hog cholera virus (HCV) was used to test the specificity of four hybridoma cell lines secreting monoclonal antibodies against pestiviruses. Reaction of virus isolates and monoclonal antibodies was controlled by an indirect immunofluorescence assay (IFA). Two monoclonal antibodies which had been generated against HC virus strain "Alfort 187" were reactive only with HCV field isolates and an HCV reference strain but not with bovine viral diarrhoea virus (BVDV) reference strains. Two other monoclonal antibodies (generated against BVDV, strain NADL) reacted only with BVDV reference strains but not with HCV field isolates, although with 3 of these strains focal reactions involving only a few cells were detected. The ability to discriminate between both viruses is a diagnostic need which may be fulfilled by these monoclonal antibodies.  相似文献   

3.
Isolates of non-cytopathogenic bovine viral diarrhoea (BVD) virus from 18 persistently infected calves from one herd were compared by using monoclonal antibodies directed against the major viral glycoprotein gp53. All the isolates displayed an almost identical reaction pattern. Based on this antigenic analysis three cytopathogenic BVD and three non-cytopathogenic BVD viruses closely related to the non-cytopathogenic BVD herd isolate were selected. Six of the persistently infected calves were inoculated with a pool of the three closely related cytopathogenic BVD viruses and two with a pool of the three non-cytopathogenic BVD viruses. In addition three animals were infected with one closely related cytopathogenic BVD strain (Indiana) and two animals with the antigenetically different cytopathogenic BVD viral strain A1138/69. Regardless of the inoculation route all the animals superinfected with closely related cytopathogenic BVD viruses developed the characteristic lesions of mucosal disease within 14 days of infection. Animals which were inoculated with non-cytopathogenic BVD viruses which closely resembled the herd isolate, or with cytopathogenic BVD viruses which did not resemble the herd isolate did not develop any signs of disease. However, the latter group produced antibodies to the superinfecting virus.  相似文献   

4.
To examine the specificity of the antibody response to the influenza hemagglutinin and the generation of antigenic variants, chickens were immunized against the highly virulent H5 virus A/Ty/Ont/7732/66 (H5N9) and then challenged with a lethal dose of the virus. The antibody responses of these chickens to the hemagglutinin (HA) were examined with an enzyme-linked immunosorbent assay (ELISA) in which their sera were titrated for the ability to block the binding of monoclonal antibodies (MAbs) to five distinct neutralizing epitopes on the viral HA. Based on the ELISA results, a majority (5/6) of the chickens produced antibodies to three of the five neutralizing epitopes on the viral HA. After challenge, two of six immunized chickens shed virus and died; antigenic comparisons of isolates from these two chickens indicated the presence of an antigenic variant; i.e., there was a change in one neutralizing epitope on the HA of virus shed by one chicken. None of the chickens had produced antibodies to this particular epitope on the viral HA. Inoculation of chickens with this variant resulted in 100% mortality, demonstrating that a change in this particular epitope did not alter the virulence of the virus. These studies indicate that chickens immunized against highly virulent influenza viruses may excrete virulent variants following challenge with live virus.  相似文献   

5.
Chickens were inoculated with infectious bursal disease virus serotype I or serotype II to determine if their immune system can distinguish between the two serotypes. Chickens had neutralizing antibodies to only serotype I viruses following exposure to serotype I viruses, and chickens had antibodies to only serotype II viruses following exposure to serotype II viruses. No cross-reactions were observed between antisera prepared to each of these two serotypes using a cross-virus-neutralization assay. Signs of disease were detected only in birds exposed to a virulent serotype I isolate. Chicks exposed to the serotype II viruses were not protected from challenge with a virulent serotype I isolate. In one experiment, antibodies to a serotype II isolate, which were detected before challenge, did not protect chicks from challenge with a virulent serotype I isolate.  相似文献   

6.
To develop a live vaccine for equine herpesvirus type 1 (EHV-1), two EHV-1 mutants containing no heterogeneous DNA, DeltagI and DeltagE, were constructed with deletions in the open reading frame of either glycoprotein I (gI) or E (gE), respectively. In equine cell culture, deletion mutants formed smaller plaques than the parental and revertant viruses, but the one-step growth patterns of the deletion mutants and the parental strain were approximately the same. These results suggest that both gI and gE contribute to the ability of EHV-1 to spread directly from cell-to-cell, but that these glycoproteins are not required for viral growth in vitro. Mice and hamsters inoculated intranasally with these mutants showed no clinical signs, and continued to gain weight, whereas those inoculated with the parental virus exhibited a reduction in mean body weight. Furthermore, nervous manifestations were observed in hamsters inoculated with the parental virus. These results suggest that gI and gE have an important role in EHV-1 virulence including neurovirulence in experimental animal models. On the other hand, serum neutralizing antibodies were detected in mice immunized with DeltagI or DeltagE at two weeks after inoculation. Following challenge with the parental virus, DeltagI- or DeltagE-immunized mice were able to clear parental virus from their lungs faster than mock-immunized mice. These results suggest that the EHV-1 mutants defective in gI and in gE are attenuated but have ability to elicit immune responses in inoculated mice that contribute to virus clearance.  相似文献   

7.
采集浙江某猪场疑似猪伪狂犬病发病仔猪的脑、脾脏等组织病料,经PCR检测为猪伪狂犬病病毒(pseudorabies virus,PRV)野毒感染,用BHK-21细胞进行病毒的分离培养,结果显示该病毒能引起典型的细胞病变,第4代病毒液毒价达107.0 TCID50/mL;PCR和动物回归试验结果表明该分离株为PRV,并将其命名为PRV ZJ株。将第4代病毒液制备成油乳剂灭活苗,免疫2 kg左右家兔,免疫后28 d采血并攻毒,测定其免疫原性,结果显示血清中和指数为13490,保护率为100%。本试验结果表明PRV ZJ株有很好的免疫原性,为进一步开展疫苗研究奠定基础。  相似文献   

8.
Monoclonal antibodies reactive to the bovine viral diarrhea virus (BVDV) protein gp53 were produced and characterized. These antibodies and our panel of anti-p80/125 monoclonal antibodies were tested for their cross-reactivity with 11 different North American and European (Danish) BVDV strains and isolates including viruses of both cytopathic and noncytopathic biotypes. The four anti-gp53 monoclonal antibodies were neutralizing for the homologous Danish cytopathic isolate and cross-reacted with all BVDV strains examined except for the Draper strain. Further, anti-gp53 monoclonal antibodies neutralized the majority of BVDV strains examined. The anti-p80/125 monoclonal antibodies cross-reacted with all eleven strains and isolates tested. This indicated that various strains of BVDV have common epitopes. The broad cross-reactivities demonstrated by these monoclonal antibodies suggest that a pool of these antibodies may be used for detection of BVDV cellular contamination or for virus isolation, in place of polyclonal antiserum.  相似文献   

9.
根据GenBank中已发表的猪伪狂犬病病毒(PRV) gE、gI基因的序列设计了2对引物,对PRV NP株的gE、gI基因进行了PCR扩增、回收、克隆、测序,测序结果与预期的PRV gE、gI基因片段相符。同源性比对分析结果显示,PRV NP株gE、gI基因推导的氨基酸序列与国内分离的PRV毒株的同源性分别为95.7%~99.8%、89.9%~99.5%。遗传进化树分析和氨基酸序列比对结果发现PRV NP株的gE氨基酸序列发生变化的位点与2012年国内分离到的PRV流行株相同,从而推测NP株为PRV变异毒株,本研究为PRV的流行病学调查分析奠定了基础,也为开发科学、有效的新型猪伪狂犬病(PR)疫苗提供科学依据。  相似文献   

10.
To determine histopathological damage in the respiratory tract, ducks were inoculated with five different influenza A viruses, including viruses virulent for other avian hosts. Lungs were collected for detection of virus and histopathological examination. Small amounts of infectious virus were recovered from lungs, and viral antigens were demonstrated by immunoperoxidase staining with monoclonal antibodies to the viral nucleoprotein. Although clinical signs were not detected, lungs of ducks infected with both virulent and avirulent viruses had mild pneumonia characterized by infiltrates of lymphocytes and macrophages. These findings show that although clinical signs are not evident, ducks may have damage to the respiratory tract during influenza.  相似文献   

11.
Pestiviruses were isolated from seven cases of suspect hog cholera. Using peroxidase conjugates of monoclonal antibodies (Mabs) six isolates were identified as hog cholera viruses (HCV), while one isolate was of ruminant origin, possibly bovine viral diarrhea virus. In parallel attempts were made to develop an ELISA for the detection of HCV-specific antibodies in pig sera. The Mab HCTC26 coated to polystyrol plates efficiently captured the major viral glycoprotein gp53 from crude antigen suspensions prepared from infected cells. The immobilized gp53 served as diagnostic antigen. Five pigs experimentally infected with the HCV strain Glentorf were sequentially bled and the development of antibodies was monitored by neutralization tests and the ELISA. Results showed that both tests detected antibodies simultaneously after infection. Titres measured by ELISA were slightly higher than those registered by neutralization.  相似文献   

12.
Subunit pseudorabies vaccines that contained only purified glycoproteins of either of 2 strains of pseudorabies virus (PRV) were prepared and subsequently tested for safety and efficacy. The strains of virus used for vaccine production differed in at least 2 properties. One strain (Kojnok) was virulent for pigs and was believed to code for the entire complement of viral glycoproteins. The other (Kaplan) was a deletion mutant that was unable to code for structural viral glycoproteins gI and gp63. Purified glycoproteins were dispersed in an oil-in-water emulsion and were administered IM to pigs. Both vaccines were found to be safe and effective immunogens. Neither caused any local or general reactions, as verified by examination of the injection site (local safety) and by vaccination of pregnant sows in PRV-infected and noninfected herds. Sows vaccinated with the gI+ or gI- vaccine protected their pigs at levels of 93 and 92%, respectively, against a severe challenge exposure that killed 98% of pigs born from nonvaccinated sows. Vaccinated pigs were tested for active immunity by intranasal challenge exposure with the NIA 3 strain. Protection was quantitated by measuring the relative daily weight difference, expressed in percent per day, between vaccinated and control pigs during the first week after challenge exposure (delta G7); the estimated differences were 2.25 and 2.13% for gI+ and gI- vaccines, respectively. The absence of gI and gp63 did not affect the efficacy of this type of subunit glycoprotein vaccines.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

13.
This experiment was done to determine the effect(s) of single passage of pseudorabies virus in dead-end hosts on the stability of the pseudorabies virus genome. Calves, dogs, rabbits and cats were inoculated with a virulent strain of pseudorabies virus and the virus was reisolated from each animal and restriction endonuclease analysis was used to determine possible alterations in the DNA banding patterns. The restriction fragment migration profile of the pseudorabies virus DNA isolated from the animals was indistinguishable from the DNA profile of the original pseudorabies virus inoculum. The restriction endonuclease viral DNA profile appears to be relatively stable after a single passage in dead-end hosts, although minor changes in the viral genome that are not detectable in the DNA banding pattern may have occurred.  相似文献   

14.
Viruses recovered from tissues taken at necropsy from American black bears were examined by use of immunofluorescence with polyclonal and monoclonal antibodies, virus neutralization with monoclonal antibodies, and restriction endonuclease analyses of the viral genomes. With these techniques, viruses were determined to be canine adenovirus type 1. Seronegative dogs that were inoculated with the virus had clinical signs typical of infectious canine hepatitis, suggesting that the virus, which was virulent for bears, was not a vaccinal strain, but a wild strain of canine adenovirus type 1.  相似文献   

15.
The interaction between a panel of ten monoclonal antibodies and hemorrhagic enteritis virus, a group II avian adenovirus, was determined. The monoclonal antibodies reacted with all nine isolates of group II avian adenoviruses, but not with any of five types of group I avian adenoviruses. All ten monoclonal antibodies recognized antigenic determinants on the hexon protein of hemorrhagic enteritis virus when analyzed by immunoprecipitation and immunoblotting. They reacted only with the native hexon protein and not with protein denatured by sodium dodecyl sulfate or guanidine-HCl/urea treatment combined with reduction and carboxymethylation. Based on the results of competitive binding assays, the panel of monoclonal antibodies could be subdivided into two groups, which recognized different antigenic domains of the hemorrhagic enteritis virus hexon protein. The monoclonal antibodies in group 1 neutralized hemorrhagic enteritis virus infectivity while the monoclonal antibodies of group 2 did not. Group 1 consisted of eight monoclonal antibodies which could be further subdivided into subgroups 1A, 1B, 1C and 1D. The subdivision of the monoclonal antibodies was based on the degree of blocking in the competitive binding assays and differences in their ability to induce enhancement. In general, the monoclonal antibodies had a higher avidity for the virulent isolate of hemorrhagic enteritis virus than for the avirulent hemorrhagic enteritis virus isolate.  相似文献   

16.
Peste des petits ruminants (PPR) is an acute, febrile viral disease of small ruminants, caused by a virus of the genus Morbillivirus. PPR and rinderpest viruses are antigenically related and need to be differentiated serologically. In the present study, 23 mouse monoclonal antibodies were produced by polyethyleneglycol (PEG)-mediated fusion of sensitized lymphocytes and myeloma cells. Among these, two belong to the IgM class and the remaining 21 to various subclasses of IgG. The MAbs from the IgG class designated 4B6 and 4B11 neutralized PPR virus in vitro. In radioimmunoprecipitation assay, 10 MAbs recognized nucleoprotein, 4 recognized the matrix protein and one each haemagglutinin and phosphoprotein. The remaining 7 MAbs failed to precipitate any defined viral protein. The reactivity pattern of the monoclonal antibodies in indirect ELISA indicated a close antigenic relationship within three Indian PPR (lineage 4) virus isolates and also within two rinderpest vaccine strains. All PPR virus isolates could be distinguished from rinderpest vaccine viruses on the basis of the reactivity pattern of all MAbs and anti-N protein MAbs. A set of six monoclonal antibodies specific to PPR virus could also be identified from the panel. From the panel of MAbs available, two MAbs were selected for diagnostic applications, one each for the detection of antigens and antibodies to PPR virus.  相似文献   

17.
This study identifies the in vitro differences (markers) between virulent and attenuated transmissible gastroenteritis (TGE) viruses. Exposure of virulent Miller strain and attenuated Purdue strain TGE viruses to a spectrum of acidities indicated that the Miller strain was more stable at pH 2. Acidities at or above pH 3 did not reduce viral infectivity of either strain. When virulent and attenuated viruses were exposed to gastric fluids of either fed or fasted swine, there was a similar degree of sensitivity. Carboxypeptidase B, alpha-amylase, and alkaline phosphatase present in porcine small intestinal fluids did not cause a significant difference in sensitivity between virulent and attenuated virus isolates. The digestive enzymes: trypsin, alpha-chymotrypsin, pancreatin, peptidase, and carboxypeptidase A did not (or only slightly) inactivate virulent Miller strain TGE virus, but greatly reduced infectivity of attenuated viruses (Purdue strain and TGE vaccine virus isolates). The attenuated strains were significantly more sensitive to small intestinal fluids from both fasted and fed adult swine. Differential sensitivities between virulent and attenuated TGE viruses to digestive fluids from stomach and small intestine further substantiate the notion of differential susceptibility to small intestinal proteases as a correlate of viral virulence.  相似文献   

18.
伪狂犬病基因缺失疫苗株(SA215)某些生物学特性研究   总被引：3，自引：0，他引：3  
本试验测定了伪狂犬病ｇＥ－／ｇＩ－／ ＴＫ－／ ＬａｃＺ＋基因缺失疫苗株（ＳＡ２１５）的致细胞病变效应、安全性、免疫原性和免疫期等生物学特性。试验结果显示,该疫苗株能在Ｖｅｒｏ细胞上适应生长,并形成典型的蚀斑。其对１日龄仔猪、怀孕母猪、牛、羊以及家兔安全,无不良接种反应,接种动物不向体外散毒。ＳＡ２１５疫苗接种猪能抵御高剂量（１０７ＰＦＵ）Ｆａ株强毒感染,攻毒后试验猪的发热期、增重受阻天数、散毒滴度均低于Ｂａｒｔｈａ株疫苗接种猪,远远低于对照组猪。ＳＡ２１５接种猪能维持长时间的高水平中和抗体滴度,免疫期可达半年以上。试验结果表明,ＳＡ２１５株是一株安全、免疫原性好、免疫期长的疫苗株。  相似文献   

19.
Rabies isolates that had been stored between 1983 and 1997 were examined with a panel of anti-lyssavirus nucleocapsid monoclonal antibodies. Out of 56 isolates from cats and various wild carnivore species, 1 isolate of Mokola virus and 5 other non-typical rabies viruses were identified. The Mokola virus isolate was diagnosed as rabies in 1993 from a cat. Genetic analysis of this isolate suggests that it falls in a distinct subgroup of the Mokola virus genotype. The 5 non-typical rabies viruses were isolated from honey badgers (Mellivora capensis), African civets (Civettictis civetta) and an unidentified mongoose (Herpestidae). These isolates are representatives of rarely-reported wildlife-associated strains of rabies, probably maintained by the slender mongoose (Galerella sanguinea). These findings indicate that both Mokola virus and the mongoose-associated variant may be more common in Zimbabwe than is apparent from routine surveillance.  相似文献   

20.
In assay cultures, sera from African swine fever convalescent pigs inhibited infection by homologous African swine fever virus. The infection-inhibition capacity did not correspond with the virus-neutralizing capacity. The serum did not prevent infection by heterologous virulent viruses. Sera from pigs challenge inoculated with the homologous virulent virus and later with a heterologous virulent virus inhibited the infection by different heterologous virulent viruses. These sera did not interfere with the infection by pseudorabies virus. The specificity of the reaction indicated that the infection inhibition was caused by antibody.  相似文献   

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