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1.
Pseudorabies virus (PRV),an alpha-herpesvirus,has been developed as a live viral vector for animal vaccines.However,the PRV recombinant virus TK-/gE-/GP5+expressing GP5 of porcine reproductive and respiratory syndrome virus (PRRSV),based on the PRV genetically depleted vaccine strain TK-/gE-/LacZ+,scarcely stimulated the vaccinated animals to produce neutralizing antibodies against PRRSV.To develop a booster-specific immune response of such PRV recombinants,the ORF5m gene (the modified ORF5 gene having better immune responses)was substituted for the ORF5 gene and introduced into PRV TK-/gE-/LacZ+,resulting in a PRV recombinant named TK-/gE-/GPSm+,which expressed the modified GPSm protein.The recombinant virus was confirmed using PCR,Southern blotting and Western blotting.TK-/gE-/GPSm+and TK-/gE-/GP5+expressing the authentic GP5 protein were inoculated into Balb/c mice to evaluate their immune responses.The results indicated that the protecting neutralization antibodies (the 3/6 vaccinated mice obtained 1:16)and cell immune responses induced by TK-/gE-/GPSm+against PRRSV were higher than that induced by TK-/gE-/GP5+.Thus,the development of the new PRV recombinant expressing the modified GP5m protein as a candidate vaccine established the basis for the study of bivalent genetic engineering vaccines against PRRSV and PRV.  相似文献   

2.
Pseudorabies virus (PRV), an alpha-herpesvirus, has been developed as a live viral vector for animal vaccines. However, the PRV recombinant virus TK/gE/GP5+ expressing GP5 of porcine reproductive and respiratory syndrome virus (PRRSV), based on the PRV genetically depleted vaccine strain TK/gE/LacZ+, scarcely stimulated the vaccinated animals to produce neutralizing antibodies against PRRSV. To develop a booster-specific immune response of such PRV recombinants, the ORF5m gene (the modified ORF5 gene having better immune responses) was substituted for the ORF5 gene and introduced into PRV TK/gE/LacZ+, resulting in a PRV recombinant named TK/gE/GP5m+, which expressed the modified GP5m protein. The recombinant virus was confirmed using PCR, Southern blotting and Western blotting. TK/gE/GP5m+ and TK/gE/GP5+ expressing the authentic GP5 protein were inoculated into Balb/c mice to evaluate their immune responses. The results indicated that the protecting neutralization antibodies (the 3/6 vaccinated mice obtained 1:16) and cell immune responses induced by TK/gE/GP5m+ against PRRSV were higher than that induced by TK/gE/GP5+. Thus, the development of the new PRV recombinant expressing the modified GP5m protein as a candidate vaccine established the basis for the study of bivalent genetic engineering vaccines against PRRSV and PRV. Translated from Journal of Biotechnology, 2005, 21(6): 858–864 [译自: 生物工程学报]  相似文献   

3.
To evaluate the efficacy of intracerebral (IC) immunization, mice were immunized with formalin-inactivated pseudorabies virus (PRV) by either subcutaneous (SC) or IC injection, and then 10(6) plaque-forming units of PRV were introduced into the hindleg of the immunized or non-immunized mice by intramuscular injection. The antibody titer in serum was elevated and boosted by additional immunization via both the SC and IC routes, but was higher after IC immunization. Intracerebrally immunized mice were completely protected from mortality and neurological signs, whereas all the non-immunized and 80% of the subcutaneously immunized mice died after developing neurological signs. In mouse models, IC immunization is more effective at inducing a protective immune response against the transneural spread of PRV than SC immunization.  相似文献   

4.
Glycoprotein B mediates the absorption and penetration of the pseudorabies virus in the form of an immunodominant Ag, and represents a major target for the development of new vaccines. This study evaluated the efficiency of live attenuated Salmonella typhimurium SL7207 for the oral delivery of DNA vaccine encoding the pseudorabies virus glycoprotein B (pCI-PrVgB) in vivo, leading to the generation of both systemic and mucosal immunity against the pseudorabies virus Ag. An oral transgene vaccination of pCI-PrVgB using a Salmonella carrier produced a broad spectrum of immunity at both the systemic and mucosal sites, whereas the intramuscular administration of a naked DNA vaccine elicited no mucosal immunoglobulin (Ig)A response. Interestingly, the Salmonella-mediated oral transgene vaccination of the pseudorabies virus glycoprotein B biased the immune responses to the Th2-type, as determined by the IgG2a/IgG1 ratio and the cytokine production profile. However, oral vaccination mediated by Salmonella harbouring pCI-PrVgB showed inferior protection to systemic immunization against virulent pseudorabies virus infection. The expression of transgene delivered by Salmonella bacteria in antigen-presenting cells of both the systemic and mucosal-associated lymphoid tissues was further demonstrated. These results highlight the potential use of live attenuated S. typhimurium for an oral transgene pseudorabies virus glycoprotein B vaccination to induce broad immune responses.  相似文献   

5.
【背景】自2011年以来,猪伪狂犬病毒(pseudorabies virus,PRV)发生变异,经典的疫苗株已不能完全抵抗PRV变异株的感染,国内多个猪场出现伪狂犬病的暴发,PRV变异毒株开始在我国大规模流行。【目的】通过分离PRV流行变异毒株,并对其进行遗传进化和致病性分析,为PRV流行病学调查及疫苗研制提供实验数据。【方法】采集黑龙江某猪场感染PRV的脑组织病料,根据GenBank PRV gEgB保守序列设计引物,进行PCR鉴定。通过对gEgC基因进行序列测定和遗传进化分析。利用BHK-21细胞分离病毒,采用噬斑纯化方法对病毒进行纯化。通过电镜、间接免疫荧光对病毒进行鉴定,测定病毒生长曲线并进行致病性研究。【结果】经PCR和测序鉴定分离株为PRV流行株,将其命名为HLJ-01。遗传进化分析结果显示,该分离毒株与我国近几年分离的流行变异株位于同一分支;氨基酸序列分析结果显示,gE和gC存在国内流行变异株的特征序列,表明该分离毒株为流行变异株。生长曲线显示,分离株HLJ-01在感染48h时滴度最高(108.5TCID50/mL)。电镜观察结果显示,病毒颗粒直径约150nm,呈球形,有囊膜,囊膜外有放射状纤突,呈现典型PRV病毒特征。动物感染实验结果显示,107.0TCID50剂量感染组死亡率为100%;106.0TCID50剂量感染组死亡率为80%;105.0TCID50剂量感染组死亡率为60%。仔猪在接种病毒后均出现PRV感染的典型症状和病理变化,证实分离毒株对仔猪有较强致病力。【结论】分离获得一株猪伪狂犬病毒,经鉴定该分离株为流行变异株,而且具有较强的致病力,这为PRV流行病学分析及疫苗候选株的筛选奠定了基础。  相似文献   

6.
Stress is one of the important factors that induces reactivation of pseudorabies virus (PrV) in latently infected pigs. We established a murine model of latent PrV infection and examined the effects of mild stress treatment in order to demonstrate that this model simulates natural infection in the pig. Latently infected mice excreted PrV from the nasal cavity under stress treatments consisting of restraint, exposure to cold or transport. Similar reactions have been observed upon treatment with acetylcholine and dexamethasone. The present findings demonstrate that these kinds of mild stress reactivate the virus in murine latent infection models in a manner similar to the induction of latent infection in pigs in the field.  相似文献   

7.
【目的】研制猪伪狂犬病毒(PRV)和猪圆环病毒2型(PCV2)的二联活疫苗,并用猪IL-18作为免疫佐剂。【方法】将猪IL-18基因插入到质粒p GO中,获得的重组转移质粒p GO18与猪PRV弱毒HB98株DNA共转染ST细胞,并进行空斑筛选和纯化;RT-PCR和Western blot分别从转录和蛋白水平鉴定其表达情况。将重组病毒PGO18和PGO、PRV弱毒株HB98、PCV2灭活商品苗和1640细胞培养基分别免疫6周龄雌性昆明小鼠,4周后二次免疫,二免后4周用PCV2 DF强毒和PRV Min/A强毒接种小鼠。通过ELISA、血清中和试验和流式细胞术及攻毒保护试验评价重组病毒的免疫原性。【结果】获得了重组病毒PGO18,并且可在ST细胞内表达;PGO18可诱导小鼠机体产生PCV2的ELISA和PRV的中和抗体水平,刺激CD3+、CD4+、CD8+T细胞亚群的增殖,且能有效抵抗PCV2和PRV强毒攻击。【结论】IL-18基因可增强重组病毒的免疫效果,使重组病毒具有良好的免疫原性,有望成为防治PCV2和PRV的候选疫苗株。  相似文献   

8.
We studied the effect of the temporal regulation of herpes simplex virus (HSV) type 1 glycoprotein D (gD-1) expression in Ia+ epidermal cells (EC) and macrophages on virus specific immunity and protection from HSV-2 challenge. gD-1 was expressed on the surface of cells infected with a vaccinia recombinant containing gD-1 under the control of an early vaccinia virus promoter (VP176). It was not expressed in cells infected with a recombinant (VP254) in which gD-1 is controlled by a late vaccinia virus promoter. BALB/c mice immunized with both recombinants seroconverted to HSV-2 as determined by neutralization. However, HSV specific delayed type hypersensitivity (DTH) responses were significantly (p<0.025) higher in VP176 than VP254 immunized animals. Both VP176 and VP254 immunized mice were protected from severe neurological disease due to HSV-2 challenge at 14 days post immunization, but long term protection was observed only in VP176 immunized mice.  相似文献   

9.
Dobson C. & Owen M. E. 1978. Effect of host sex on passive immunity in mice infected with Nematospiroides dubius. International Journal for Parasitology8: 359–364. Female C3H but not Quackenbush (Q) mice harboured fewer Nematospiroides dubius than male C3H and Q mice. Both strains lost worms 21 days after infection. C3H and Q mice became progressively immune to infection following 4 sequential doses of N. dubius larvae and showed a sex resistance to infection. Hypothymic nu/nu CBA Balb/c mice did not show these effects on N. dubius infection. The reciprocal transfer of male and female immune mesenteric lymph node cells (IMLNC) to syngeneic male and female recipients showed that the female environment enhanced the protective qualities of both male and female IMLNC but the male environment suppressed these effects. Gonadectomized male and female recipients of male and female IMLNC had levels of infection similar to the entire female recipients. Serum from immune female donor mice protected both male and female recipients better than immune serum from male donors, but female mice in each treatment group were better protected than male mice. Immune serum transferred greater levels of protection then IMLNC to recipient mice against N. dubius infections. These data are consistent with the conclusion that the male environment suppresses lymphocyte activity and the production of protective antibodies and additionally may depress the effectiveness of sensitized lymphocytes and antibodies in ejecting N. dubius. On the other hand the female environment does not appear to adversely affect the mobilization of the protective immune response and may enhance immune effector mechanisms in protecting mice against N. dubius infections.  相似文献   

10.
Nectin-1 is a Ca2+-independent Ig-like cell–cell adhesion molecule and an alphaherpesvirus receptor that binds to virion glycoprotein D by the first Ig-like domain. We have investigated the antiviral potentials of soluble forms of porcine nectin-1 to PRV infection by generating transgenic mice expressing different types of fusion protein. Previously, we reported that mice transgenic for a chimera that carried the entire ectodomain of porcine nectin-1 fused to the Fc portion of porcine IgG1 were more resistant than those transgenic for a chimera that carried the first Ig-like domain fused to the Fc portion. Recently, we generated transgenic mice expressing a fusion protein made of the first Ig-like domain fused to the Fc portion of human IgG1, and reported that they showed a microphthalmia. Here, two transgenic mouse lines expressing the fusion protein were challenged with PRV for comparing their resistances with those of transgenic mice expressing different types of fusion protein. Surprisingly, both transgenic mouse lines showed a high resistance to the viral infection, especially via the i.n. route. Significant resistance of the embryonic fibroblasts was also observed. Altogether, these findings indicated that the fusion protein consisting of the first Ig-like domain fused to the human Fc portion provided a marked resistance against PRV infection to the transgenic mice.  相似文献   

11.
Hong Q  Qian P  Li XM  Yu XL  Chen HC 《Biotechnology letters》2007,29(11):1677-1683
Pseudorabies (PR), foot-and-mouth disease (FMD), and porcine parvovirus disease are three important infectious diseases in swine worldwide. The gene-deleted pseudorabies virus (PRV) has been used as a live-viral vector to develop multivalent genetic engineering vaccine. In this study, a recombinant PRV, which could co-express protein precursor P1-2A of FMDV and VP2 protein of PPV, was constructed using PRV TK/gE/LacZ+ mutant as the vector. After homologous recombination and plaque purification, recombinant virus PRV TK/gE/P1-2A-VP2 was acquired and identified. Immunogenicity, safety of the recombinant PRV and its protection against PRV were confirmed in a mouse model by indirect ELISA and serum neutralization test. The results show that the recombinant PRV is a candidate vaccine strain to develop a novel trivalent vaccine against PRV, FMDV and PPV in swine.  相似文献   

12.
目的:探讨玩具对实验小鼠生长发育及繁殖性能的影响。方法选取7-8周龄ICR小鼠,在生产过程中加入玩具进行干预,观测其对小鼠生长发育及繁殖性能方面指标的影响。结果除产仔数外,实验组各项繁殖性能指标均优于对照组,且差异显著( P <0.05)。并且随干预时间延长,差异的显著性有进一步增加的趋势。实验组小鼠的生长发育水平相比于对照组有明显变化,且表现出有随干预时间延长而产生差异的趋势,但目前尚无统计意义( P >0.05)。结论玩具对ICR小鼠的生长发育及繁殖性能有一定程度的积极影响,在小鼠的生产和饲育过程中值得借鉴。  相似文献   

13.
ABSTRACT The potential advantages of repeated breeding at a particular location should improve reproductive performance in long‐lived species of birds. However, for short‐lived species, natural selection should favor individuals that most quickly develop competency in reproduction. Therefore, we hypothesized that local breeding experience beyond the first breeding attempt at a particular location would have little effect on subsequent reproductive performance of Tree Swallows (Tachycineta bicolor), a species where about 50% of adults breed only once in their lives. We tested this hypothesis using data collected from Tree Swallows in Michigan from 1993 to 2002. Because we were specifically interested in examining the effects of local breeding experience on reproductive performance, we restricted our analyses to after‐second‐year (ASY) females and their mates that we first encountered as breeders. Consistent with our hypothesis, we found no relationship between repeated local breeding experience and the reproductive performance of ASY female Tree Swallows and their mates as measured by clutch size and number of fledged young. However, pairs with more combined total local experience tended to lay eggs earlier in the season. These results suggest that Tree Swallows may benefit from breeding site fidelity, not because repeated local experience improves reproductive performance as measured by the production of fledglings, but because returning individuals acquire nest cavities earlier and are able to begin breeding earlier, providing time to renest in case of early nest failure.  相似文献   

14.
Pseudorabies, a herpesvirus infection, is mainly controlled by using attenuated live vaccines. In this study, the effect of ginseng stem and leaf saponins (GSLS) in combination with selenium (Se; in the form of sodium selenite) on vaccination against attenuated pseudorabies virus (aPrV) was evaluated. It was found that GSLS and Se have an adjuvant effect and that a combination of GSLS and Se stimulates significantly enhanced immune responses than does GSLS or Se alone. Following oral administration of GSLS, mice immunized with an attenuated PrV vaccine diluted in Se‐containing physiological saline solution (PSS) provoked a significantly stronger gB‐specific serum antibodies response (IgG, IgG1 and IgG2a), enhanced lymphocyte proliferation and cytolytic activity of NK cells, along with higher production of cytokines (IFN‐γ, IL‐12, IL‐5 and IL‐10) by splenocytes. Notably, the combination of GSLS and Se conferred a much higher resistance to fPrV challenge after immunization of the mice with aPrV vaccine. This study offers convincing experimental evidence that an injection of Se with oral GSLS is a promising adjuvant combination that improves the efficacy of vaccination against PrV and deserves further study regarding improvement of responses to other animal vaccines.  相似文献   

15.
Herpes simplex virus (HSV) is common throughout the world and is a target for vaccine development. Transcutaneous immunisation is a novel technique that uses the application of vaccine antigens in solution on the skin in the presence of cholera toxin (CT) as an adjuvant. This study investigated the potential of transcutaneous immunisation in C3H mice, using CT co-administered with whole inactivated HSV-1 (CT+HSVi) or HSV-1 antigens extracted from infected Vero cells (CT+HSVag) or a control protein (CT+BSA). The application of any of the three vaccines on to bare mouse skin resulted in the migration of Langerhans cells from the epidermis and in the production of serum antibodies to CT. Both HSV preparations generated serum and mucosal (faecal) antibodies to HSV, with the CT+HSVi vaccine being a more potent stimulator of humoral immunity. The CT+HSVag vaccine, however, was the more potent stimulator of cell-mediated immunity, giving rise to a strong delayed type hypersensitivity response and lymphocyte proliferation in vitro. When the mice were challenged by epidermal inoculation of HSV, the CT+HSVag vaccine induced a higher level of protection than the CT+HSVi vaccine, a result which may indicate that the efficacy of HSV vaccines depends on stimulation of cell-mediated rather than humoral responses. The success of topical vaccination suggests that the transcutaneous route may offer a promising potential for novel vaccine delivery which merits further investigation.  相似文献   

16.
The effect of dose of pregnant mares' serum gonadotropin (PMSG) on the reproductive performance of adult ewes and ewe lambs and lamb survival at birth after treatment with fluorogestone acetate (FGA)-impregnated intravaginal sponges and PMSG (250 IU or 500 IU) to synchronize estrus was evaluated. Ewes were exposed to rams for breeding at the synchronized and subsequent estrous cycles. The flock, comprised of three synthetic strains and two control breeds, was maintained in a controlled environment and exposed to an artificial light regimen which alternated at 4-mo intervals from 16h of light daily to 9h of light daily. Trials were conducted during January, May and September at the end of a 9-h daylength cycle. Adult ewes were bred in May and 8 mo later in January. Ewe lambs were bred in September at 6.5 to 7.5 mo of age. The overall reproductive performance of the adult ewes was similar at the two breedings: fertility approximately 90%, prolificacy approximately 2.7, fecundity approximately 240% and lambs born alive approximately 2.4. Dosage of PMSG had no effect. Reproductive performance of ewe lambs was lower and there was a strain x treatment interaction, suggesting greater variability in response. The results indicate there is no advantage to using a higher dose of PMSG in ewes with a natural relatively high fecundity. Moreover, the use of the artificial photoperiod appears to overcome the natural seasonal variation in reproductive performance.  相似文献   

17.
Human respiratory syncytial virus (RSV) is a serious pediatric pathogen of the lower respiratory tract worldwide. There is currently no clinically approved vaccine against RSV infection. Recently, it has been shown that a replication-deficient first generation adenoviral vector (FGAd), which encodes modified RSV attachment glycoprotein (G), elicits long-term protective immunity against RSV infection in mice. The major problem in developing such a vaccine is that G protein lacks MHC-I-restricted epitopes. However, RSV fusion glycoprotein (F) is a major cytotoxic T-lymphocyte epitope in humans and mice, therefore, an FGAd-encoding F (FGAd-F) was constructed and evaluated for its potential as an RSV vaccine in a murine model. Intranasal (i.n.) immunization with FGAd-F generated serum IgG, bronchoalveolar lavage secretory IgA, and RSV-specific CD8+ T-cell responses in BALB/c mice, with characteristic balanced or mixed Th1/Th2 CD4+ T-cell responses. Serum IgG was significantly elevated after boosting with i.n. FGAd-F. Upon challenge, i.n. immunization with FGAd-F displayed an effective protective role against RSV infection. These results demonstrate FGAd-F is able to induce effective protective immunity and is a promising vaccine regimen against RSV infection.  相似文献   

18.
As a result of the increase in herd size and the intensification of production, the complexity of reproductive management has been growing in dairy herds. The aim of our study was to examine the associations of management practices and reproductive performance in Holstein cows on large commercial dairy farms. Management practices applied to cows were surveyed between 22 May and 6 November 2015 in 34 large Holstein-Friesian dairy herds in Hungary. Individual data of 23 784 cows that calved between 1 January 2014 and 31 December 2014 in the surveyed herds were gathered. Associations between the management practices and the reproductive parameters were analyzed by mixed effects models. Regarding heat abatement we found that ventilation with sprinklers was associated with the shortest breeding interval (P<0.01), the shortest calving to conception interval (P<0.01), and the highest odds of being pregnant by 200 days in milk (P<0.01). Solely ventilation showed similar (P>0.05) results to lack of heat stress protection. It was also revealed, that lack of a well-established voluntary waiting period (VWP) or a VWP shorter than 50 days was associated with less days to first service (P<0.01), shorter breeding interval (P<0.01) and calving to conception interval (P<0.05), as well as higher odds of carrying a calf by 200 days in milk (P<0.01) compared with those using a VWP of at least 50 days. Those farms that applied transrectal ultrasonography were more likely to use ventilation with sprinklers (P<0.05), hormonal synchronization (P<0.01) and to perform early pregnancy diagnosis followed by pregnancy recheck (P<0.05). The application of transrectal ultrasonography with one of the aforementioned practices was associated with reduced days to first service (P<0.05), shorter breeding interval (P<0.05) and higher odds of pregnancy at 200 days in milk (P<0.05). Our study highlights the management practices most closely related to improved reproductive performance, which are, therefore, suggested to be applied on dairy farms, considering the local circumstances of the individual farms.  相似文献   

19.
Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of PRRS, which causes severe reproductive failure in sows, respiratory disease in young and growing pigs, and enormous economic losses to the global swine industry. In this study, SILAC combined with MS/MS was used to quantitatively identify the secretory proteins differentially expressed in PRRSV‐infected Marc‐145 cells compared with mock‐infected controls. In total, we identified 204 secretory proteins showing significant differences in infected cells (163 upregulated, 41 downregulated). Intensive bioinformatic analysis of secretome data revealed that PRRSV infection strongly activated nonclassical protein secretion, especially vesicle‐mediated release of exosomal proteins, including different danger‐associated molecular pattern molecules and the majority of secreted proteins involved in protein binding and transport, regulation of response to stimulus, metabolic processes, and immune responses. According to the functional proteins analysis, we speculate that proteins functioning in binding, transport, and the immune response are exploited by PRRSV to facilitate virus replication and immune evasion. Our study for the first time analyzes the secretory protein profile of PRRSV‐infected Marc‐145 cells and provides valuable insight into the host response to PRRSV infection.  相似文献   

20.
Infection can cause hosts to drastically alter their investment in key life‐history traits of reproduction and defence. Infected individuals are expected to increase investment in defence (e.g., by increasing immune function) and, due to trade‐offs, investment in other traits (e.g., current reproduction) should decrease. However, the terminal investment hypothesis postulates that decreased lifespan due to infection and the associated reduction in the expectation for future offspring will favour increased investment towards current reproduction. Variation in intrinsic condition will likely influence shifts in reproductive investment post‐infection, but this is often not considered in such assessments. For example, the extent of inbreeding can significantly impact an individual's lifetime fitness and may influence its reproductive behaviour following a threat of infection. Here, we investigated the effects of inbreeding status on an individual's reproductive investment upon infection, including the propensity to terminally invest. Male crickets (Gryllodes sigillatus) from four genetically distinct inbred lines and one outbred line were subjected to a treatment from an increasing spectrum of simulated infection cue intensities, using heat‐killed bacteria. We then measured reproductive effort (calling effort), survival and immune function (antibacterial activity, circulating haemocytes and haemocyte microaggregations). Inbred and outbred males diverged in how they responded to a low‐dose infection cue: relative to unmanipulated males, outbred males decreased calling effort, whereas inbred males increased calling effort. Moreover, we found that inbred males exhibited higher antibacterial activity and numbers of circulating haemocytes compared with outbred males. These results suggest that an individual's inbreeding status may have consequences for context‐dependent shifts in reproductive strategies, such as those triggered by infection.  相似文献   

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