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表达新城疫病毒融合蛋白基因禽痘病毒重组体的构建 总被引：1，自引：0，他引：1
通过RT-PCR扩增新城疫病毒F基因,利用禽痘病毒复合启动子、SV40 PolyA终止信号序列构建F基因表达盒。将F基因表达盒与loxp-GFP-loxP序列串联插入禽痘病毒重组臂,构建了禽痘病毒转移质粒载体。在脂质体介导下转染CEF,获得了带有绿色荧光信号的重组病毒rFPVFGFP。通过二次转染,利用Cre-loxP位点特异性重组将重组病毒基因组的GFP自动去除,结果获得了只含新城疫病毒F基因表达盒的重组禽痘病毒rFPVF。试验结果表明,rFPVF复制稳定,表达的F蛋白具有免疫原性,能够刺激鸡只产生抗新城疫病毒的中和抗体。 相似文献
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禽痘病毒感染对禽流感重组禽痘病毒疫苗免疫效力的影响 总被引：1，自引：0，他引：1
表达禽流感病毒 (AIV)HA和NA基因的重组禽痘病毒rFPV_HA_NA能够诱导鸡体产生 10 0 %抵抗高致病性禽流感病毒 (HPAIV)H5N1的攻击。而当鸡群已进行禽痘疫苗免疫或者感染了禽痘病毒的情况下 ,此重组疫苗的免疫效力如何 ?首先用禽痘病毒S_FPV_0 17人工感染SPF试验鸡 ,既而在感染后的不同间隔时间接种重组疫苗 ,免疫后检测鸡群的HI抗体水平 ,同时用 10 0LD50 的HPAIVH5N1进行攻击。结果重组疫苗免疫与禽痘病毒人工感染时间间隔在 4周 (或以上 )时 ,预先感染禽痘病毒对重组疫苗的免疫效力不构成影响 ,对禽流感的保护力为 10 0 % ,而间隔时间在 1、2、3周时 ,重组疫苗的免疫保护效力则受到不同程度的影响。 相似文献
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鸡新城疫的流行与免疫 总被引：1，自引：0，他引：1
门诊病例统计表明，我国鸡群中ND的发病率仍非常普遍。近些年来，世界各地对NDV分离株的研究结果发现，其HN基因和F基因序列的某些位点发生的变化，致使毒株间的毒力差异较大，抗原性亦有差异，但其主要抗原表位未发生变化。山东家禽研究所禽病室对1998－1999年的6个NDV分离株做了比较研究，各毒株的毒力不同且都比经典的F48E9强，其中二个属基因Ⅶ型。但交叉免疫保护试验表明，用La Sota株弱毒苗及其灭活油乳苗免疫的鸡，在用这6个分离毒分别攻毒后，均有1005保护率。同样，用这6个分离毒制成的灭活油乳苗免疫鸡后，对经典毒E48F9亦产生100％保护率。这表明，从目前发病鸡群中分离的NDV毒株与La Sota弱毒株、标准F48E9强毒株间有很好的交叉免疫保护作用。 相似文献
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为评价共表达鸡IL-6和H5亚型禽流感病毒HA基因重组鸡痘病毒(rFPV-AIH5 AIL6)的免疫抗体消长规律及免疫效力,将重组病毒通过颈部皮下注射和翅部皮下注射2种不同的免疫途径来免疫鸡群,结果发现,2种免疫途径中,重组鸡痘病毒对鸡体质量增加均无影响,而野生型鸡痘病毒均可抑制鸡体质量增加.翅部皮下注射疫苗组能够产生较高的血凝抑制(HI)抗体水平,免疫21 d后抗体水平达到高峰,28 d后开始下降,49 d时仍保持在一定的水平.免疫SPF鸡21 d后攻毒,表明该重组病毒能使经滴鼻攻毒的SPF鸡抵抗H5亚型AIV的致死性攻击,保护率为95％,与油苗组相同,与单表达H5亚型禽流感病毒HA基因重组鸡痘病毒组(40％)差异显著;攻毒后3、5、7d采集喉头、泄殖腔棉拭子检测排毒情况,结果发现第3天排毒率最高,其中rFPV-AIH5AIL6免疫组排毒率为最低,显示IL-6在rFPV-AIH5IL6免疫过程中起到了免疫佐剂的作用,这为研制新型的禽流感重组鸡瘟病毒疫苗奠定了基础. 相似文献
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为评价共表达鸡IL-6和H5亚型禽流感病毒HA基因重组鸡痘病毒(rFPV-AIH5AIL6)的免疫抗体消长规律及免疫效力,将重组病毒通过颈部皮下注射和翅部皮下注射2种不同的免疫途径来免疫鸡群,结果发现,2种免疫途径中,重组鸡痘病毒对鸡体质量增加均无影响,而野生型鸡痘病毒均可抑制鸡体质量增加。翅部皮下注射疫苗组能够产生较高的血凝抑制(HI)抗体水平,免疫21d后抗体水平达到高峰,28d后开始下降,49d时仍保持在一定的水平。免疫SPF鸡21d后攻毒,表明该重组病毒能使经滴鼻攻毒的SPF鸡抵抗H5亚型AIV的致死性攻击,保护率为95%,与油苗组相同,与单表达H5亚型禽流感病毒HA基因重组鸡痘病毒组(40%)差异显著;攻毒后3、5、7d采集喉头、泄殖腔棉拭子检测排毒情况,结果发现第3天排毒率最高,其中rFPV-AIH5AIL6免疫组排毒率为最低,显示IL-6在rFPV-AIH5IL6免疫过程中起到了免疫佐剂的作用,这为研制新型的禽流感重组鸡痘病毒疫苗奠定了基础。 相似文献
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鸡新城疫病毒分离株与La Sota株灭活疫苗效力比较试验 总被引：1，自引：0，他引：1
用NDV分离株及La Sota株为抗源液，经福尔马林灭活后，与油佐剂混合，分别制成分离株灭活苗、La Sota株灭活苗及分离株与La Sota株二价灭活苗。将这三种灭活疫苗分别免疫SPF鸡后，均获得100%抵抗NDV分离株及F48株强毒攻击的保护力；而用这3种灭活苗与La Sota活苗单独或联合使用，免疫带有ND母源抗体的普通鸡后，3种灭活苗的免疫效力不同，分离株灭活苗与价灭活苗对NDV分离株攻击的免疫保护效力明显优于La Sota灭活苗；灭活苗与活苗同时使用，其免疫效力明显优于单独使用灭活苗或活苗。 相似文献
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以鸡毒支原体NB72株或和R株对鸡先行人工感染,然后,与无支原体感染的对照鸡同样接种NDV La Sota株,观察NDV免疫应答动态。结果,支原体感染鸡群血清和气管中的NDV血凝抑制抗体效价,虽然比未接种支原体的鸡群低.但二者差异不显著(P>0.05).随后,通过攻击NDV强毒,不论先前经过或未经过支原体人工感染,ND免疫鸡全部被保护,且血清中NDV HI抗体效价均大幅度升高;未经ND免疫接种的对照鸡全部发病死亡,因此,鸡毒支原体NB72株或R株感染的鸡对NDV La Sota株的免疫应答无明显改变。最后就体内试验的结果进行了讨论。 相似文献
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为研究重组鸡γ-干扰素(Chicken interferonγ, ChIFN-γ)对鸡新城疫、禽流感(H9亚型)二联灭活苗(La Sota株+WD株)的免疫增强效果,取重组鸡γ-干扰素作为免疫增强剂,联合鸡新城疫、禽流感(H9亚型)二联灭活疫苗免疫SPF鸡,在免疫后24h和48h采集全血进行细胞因子检测,结果显示免疫后联合免疫组的IFN-γ和IL-4细胞因子水平要高于单独免疫疫苗组;免疫后7、14、21、28和35d分别采集血清测定抗体效价,通过检测血清中针对鸡新城疫病毒和禽流感病毒(H9亚型)的HI抗体水平可知,联合免疫组产生的针对鸡新城疫病毒和禽流感病毒(H9亚型)的抗体均高于单独免疫疫苗组。试验结果表明,重组鸡γ-干扰素对鸡新城疫、禽流感(H9亚型)二联灭活苗(La Sota株+WD株)具有免疫增强作用。 相似文献
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将鸡传染性喉气管炎病毒（ILTV）gD基因和鸡白细胞介素2（ChIL-2）基因通过同源重组法重组禽痘病毒转移载体,构建含ILTVgD基因和ChIL-2基因的重组禽痘病毒转移载体rFPVIL2ILTV—gD,蓝色蚀斑纯化法纯化重组病毒,用IFA检测重组病毒中ILTVgD基因的表达和用ELISA试剂盒检测ChIL-2的表达。重组病毒rFPV—IL2-ILTV-gD免疫试验鸡,间接ELISA测定血清中ILTV抗体效价,动物试验用构建的ILTV强毒以滴鼻方式攻击各组试验鸡,结果表明,外源基因在重组禽疽病毒中得到了稳定表达;ELISA检测结果表明在免疫7d后能够检测到血清抗体,免疫21d后血清抗体达到高峰,此后便开始逐渐下降,符合疫苗免疫的消长规律;攻毒后每组鸡的发病情况可以看出重组病毒rFPV-IL2-ILTV-gD组和疫苗对照组对ILTV强毒攻击的保护率分别为96％和92％,而空白对照组为8％。说明构建的重组病毒rFPV-IL2-ILTV—gD免疫效力稍优于弱毒疫苗,能够较好的保护免疫鸡群。 相似文献
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R W Morgan J Gelb C S Schreurs D Lütticken J K Rosenberger P J Sondermeijer 《Avian diseases》1992,36(4):858-870
Recombinant strains of herpesvirus of turkeys (HVT) were constructed that contain either the fusion protein gene or the hemagglutinin-neuraminidase gene of Newcastle disease virus (NDV) inserted into a nonessential gene of HVT. Expression of the NDV antigens was regulated from a strong promoter element derived from the Rous sarcoma virus long terminal repeat. Recombinant HVT strains were stable and fully infectious in cell culture and in chickens. Chickens receiving a single intra-abdominal inoculation at 1 day of age with recombinant HVT expressing the NDV fusion protein had an immunological response and were protected (> 90%) against lethal intramuscular challenge at 28 days of age with the neurotropic velogenic NDV strain Texas GB. Recombinant HVT expressing the NDV hemagglutinin-neuraminidase provided partial protection (47%) against the same challenge. Chickens vaccinated with recombinant HVT vaccines had low levels of protection against NDV replication in the trachea when challenged ocularly. Recombinant HVT vaccines and the parent HVT strain provided similar levels of protection to chickens challenged with the very virulent RB1B strain of Marek's disease virus, indicating that insertion of foreign sequences into the HVT genome did not compromise the ability of HVT to protect against Marek's disease. 相似文献
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Development of a virosome vaccine for Newcastle disease virus 总被引：7，自引：0，他引：7
In an effort to protect chickens against Newcastle disease (ND), a nonreplicating virosome vaccine was produced by solubilization of Newcastle disease virus (NDV) with Triton X-100 followed by detergent removal with SM2 Bio-Beads. Biochemical analysis indicated that the NDV virosomes had similar characteristics as the parent virus and contained both the fusion and hemagglutinin-neuraminidase proteins. To target the respiratory tract, specific-pathogen-free chickens were immunized intranasally and intratracheally with the NDV virosome vaccine. This vaccine was compared with a standard NDV (LaSota) live-virus vaccine for commercial poultry. Seroconversion (> or = four fold increase in hemagglutination inhibition [HI] antibody titers) was achieved in all birds vaccinated with the virosome vaccine. Upon lethal challenge with a velogenic NDV strain (Texas GB), all birds receiving either vaccination method were protected against death. Antibody levels against NDV, as determined by enzyme-linked immunosorbent assay and HI titer, were comparable with either vaccine and increased after virus challenge. These results demonstrate the potential of virosomes as an effective tool for ND vaccination. 相似文献
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新城疫病毒(NDV)的HN蛋白是一个多功能蛋白,具有血凝素(HA)和神经氨酸酶(NA)两种活性,在病毒感染过程中扮演重要角色。本研究利用反向遗传操作技术,将NDV强毒株F48E9的HN基因替换弱毒株rLaSota的HN基因,获得嵌合病毒rL-F48E9HN。收获病毒尿囊液,提取基因组RNA,进行序列分析,结果显示嵌合病毒基因组的HN基因获得了正确替换。嵌合病毒在鸡胚内的生长特性与亲本株LaSota一致,与F48E9的生长特性相差甚远。嵌合病毒红细胞吸附性明显增强,较rLaSota株增加51%,而细胞融合作用无明显变化。rL-F48E9HN的鸡胚平均致死时间(MDT)为148 h,脑内致病指数(ICPI)为0.43,静脉接种致病指数(IVPI)为0,说明rL-F48E9HN的毒力仍然属于弱毒力范围,而没有达到中等毒力或者强毒力。 相似文献
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King DJ 《Avian diseases》1999,43(4):745-755
Four-week-old specific-pathogen-free white rock chickens were immunized with either a commercial recombinant fowl poxvirus-vectored Newcastle disease vaccine (FPN) expressing the hemagglutinin-neuraminidase and fusion protein genes of Newcastle disease virus (NDV) strain B1 or live NDV B1. Vaccinates and controls were challenged by eyedrop and intranasal (E/I) route with a viscerotropic velogenic NDV at 14 days postvaccination to determine the time of clearance of challenge virus. In a subsequent experiment, chickens were challenged at 3, 6, or 10 days postvaccination to determine the onset of immunity. Chickens that received a recommended field dose (1x) or a 0.01x dose of FP-N subcutaneously (s.c.) and were seropositive by hemagglutination-inhibition test at 14 days postvaccination cleared the challenge virus by 14 days postchallenge. Clinical Newcastle disease and high challenge virus titers in tissues were seen only in seronegative FP-N 0.01x dose vaccinates and controls. In a comparison of vaccination with FP-N (1x, 10(4,9) median tissue culture infective dose) s.c., B1 (10(6) median egg infective dose [EID50]) s.c., or B1 (10(6) EID50) E/I, chickens vaccinated at 6 or 10 days before challenge with all vaccines were protected against clinical disease, but only those vaccinated with B1 E/I 10 days before challenge were protected against infection with the challenge virus. Vaccination at 3 days before challenge with B1 E/I provided early protection, but severe nervous signs developed later and reduced overall protection to 60%, whereas disease in chickens vaccinated with B1 s.c. and FP-N s.c. 3 days before challenge was similar to the challenge controls. 相似文献
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Protective immunity against Newcastle disease: the role of antibodies specific to Newcastle disease virus polypeptides 总被引：3，自引：0，他引：3
Studies were performed to determine if passive immunization with hyperimmune sera generated to specific Newcastle disease virus (NDV) proteins conferred protection against virus challenge. Six groups of 3-wk-old chickens were passively immunized with antiserum against either hemagglutinin-neuraminidase/fusion, (HN/F) protein, nucleoprotein/phosphoprotein (NP/P), Matrix (M) protein, a mixture of all NDV proteins (ALL), intact ultraviolet-inactivated NDV (UVNDV), or negative sera. Blood samples were collected 2 days postimmunization, and the birds were challenged with Texas GB strain of NDV. Antibody titers were detected from those recipient birds that had received the antisera against the HN/F, ALL, or UVNDV by a hemagglutination inhibition test, an enzyme-linked immunosorbent assay (ELISA), and a virus neutralization test. Antibodies were detected only by the ELISA from the birds that had received antisera against NP/P and M protein. Antibody titers in the recipient birds dropped by two dilutions (log2) after 2 days postinjection. Birds passively immunized with antisera against HN/F, ALL, and UVNDV were protected from challenge, whereas chickens passively immunized with antisera against NP/P and M protein and specific-pathogen-free sera developed clinical signs of Newcastle disease. The challenge virus was recovered from the tracheas of all passively immunized groups. The presence of neutralizing antibodies to NDV provided protection from clinical disease but was unable to prevent virus shedding from the trachea. 相似文献
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Masaji MASE 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2022,84(1):1
Recently, genotype VII of Newcastle disease virus (NDV) has become the most prevalent NDV genotype in Asia. Here the hemagglutinin-neuraminidase (HN) gene of genotype VII NDV strains isolated in Japan was analyzed. Notably, point amino acid substitutions in the HN protein at position 347, which is located on the major linear epitope of the HN protein, were found in two strains. However, by a hemagglutination inhibition assay, major antigenic differences did not exist between the studied strains. Additionally, chickens vaccinated with the B1 strain did not exhibit clinical effects after challenge with variants possessing the substitution at position 347 (E to K), whereas all unvaccinated chickens subjected to this challenge died within 5 days. 相似文献
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鸡新城疫重组鸡痘病毒活疫苗的免疫持续期和加强免疫研究 总被引：1，自引：0，他引：1
本研究旨在评价表达新城疫病毒（NDV）血凝素-神经氨酸酶（HN）基因的重组鸡痘病毒（rFPV-12LSHN）活疫苗的免疫持续期和加强免疫对疫苗免疫效力的影响。用rFPV-12LSHN活疫苗免疫14日龄SPF鸡,103PFU/羽,7d即可检测到NDV HI抗体应答,对NDV强毒F48E8株攻毒保护率达100%。一次免疫18周后,对NDV强毒攻击依然提供完全保护。鸡痘病毒（FPV）疫苗免疫4周,再接种rFPV-12LSHN活疫苗,攻毒保护率降低至50%。相反,rFPV-12LSHN免疫4周,随后二次免疫可显著提高对NDV的体液免疫应答水平（P〈0.01）,对NDV强毒攻击的保护率仍然为100%。结果表明,表达NDV HN基因的重组鸡痘病毒（rFPV-12LSHN）活疫苗,能够快速建立坚强免疫力,免疫持续期至少可达18周,rFPV-12LSHN的二次免疫可以提高疫苗的免疫力。 相似文献