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1.
猪α干扰素对猪圆环病毒2型亚单位疫苗免疫效果的影响 总被引：2，自引：0，他引：2
为了评估酵母表达的PCV2 Cap蛋白在猪体的免疫特性以及猪α干扰素对其的免疫佐剂效果,将酵母表达的PCV2 Cap蛋白配比适量的铝胶或重组猪α干扰素制成亚单位疫苗.30日龄仔猪分别接种铝胶佐剂亚单位疫苗和猪α干扰素佐剂亚单位疫苗,同时设攻毒对照与空白对照,首免后21 d加强免疫,二免后14 d用PCV2 JS株攻毒.铝胶佐剂亚单位疫苗免疫猪体后产生了PCV2特异性中和抗体.猪α干扰素佐剂亚单位疫苗组仔猪免疫后产生的ELISA抗体和中和抗体都显著高于铝胶亚单位疫苗组仔猪.攻毒对照组的4只仔猪攻毒后全都产生了病毒血症,并有较长时间的发热;铝胶亚单位疫苗组4头仔猪中只有1头仔猪产生病毒血症,添加猪α干扰素亚单位疫苗组的仔猪攻毒后都没有产生病毒血症.综合分析试验猪的病毒感染、临床表现和生产性能等情况表明Cap蛋白亚单位疫苗具有免疫保护效果,猪α干扰素可显著增强Cap蛋白亚单位疫苗接种仔猪的体液免疫反应,提高PCV-2 Cap蛋白亚单位疫苗免疫保护效果. 相似文献
2.
为了对制备的猪圆环病毒2型(porcine circovirus type 2,PCV2)灭活疫苗进行安全性及免疫效力的评价,本试验将3批次猪圆环病毒2型灭活疫苗以2倍剂量接种PCV2阴性仔猪后,进行了安全性分析抗体检测和攻毒试验。结果显示：3批次疫苗以2倍剂量接种仔猪后,仔猪均无异常临床反应,可达到1∶1600以上,而攻毒对照组及空白对照组仔猪的抗体水平均在1∶400以下;攻毒后免疫组可以达到100%保护,攻毒对照组出现100%发病。结果表明,所制备的猪圆环病毒2型灭活疫苗对仔猪不仅安全性好,而且可以为仔猪提供有效的保护,为疫苗推广奠定了重要基础。 相似文献
3.
【目的】筛选更安全高效的猪圆环病毒2型(Porcine circovirus type 2,PCV2)合成肽疫苗佐剂。【方法】将IMS 251C VG、GEL 02 PR、ISA 15A VG、ISA 206 VG、ISA 61 VG、台湾细菌鞭毛、卡波姆、多糖共8种佐剂与适量PCV2多肽抗原按照特定方法配制成不同疫苗,并验证其物理性状,采用小鼠试验初步检验疫苗安全性及免疫效果,筛选出较优的佐剂进行仔猪免疫攻毒保护试验,进一步评估各疫苗的免疫保护效果。【结果】物理性状显示,所有疫苗稳定性均较好,4℃保存期均超过12个月。ISA 61 VG油佐剂黏度较高且乳滴粒径较大,ISA 15A VG、ISA 206 VG双相佐剂次之,其余水佐剂均较低较小。小鼠试验结果显示,除台湾细菌鞭毛佐剂外其他佐剂安全性均良好;小鼠ELISA抗体水平检测结果显示,ISA 61 VG、GEL 02 PR、ISA 206 VG组免疫效果明显高于其余疫苗组;综合评定安全性和免疫效果,筛选ISA 61 VG、GEL 02 PR、卡波姆、多糖、ISA 206 VG共5个佐剂组进行仔猪免疫攻毒保护试验。仔猪免疫攻毒保护试验... 相似文献
4.
为筛选满足猪圆环病毒2型灭活疫苗制苗要求的毒株,选择河南省8个地区的8株猪圆环病毒2型分离株进行抗原性分析,将各分离株培养后用不同滴度的病毒原液和其分别稀释至同一滴度(4.233×104拷贝/μL)的病毒液分别制成油佐剂灭活疫苗,免疫小鼠后采血测定其抗体动态水平,以进行免疫效力评价。结果显示,8株圆环病毒2型分离株中8#和12#分离株原液制备的疫苗免疫后诱导产生的抗体水平较高,二免后1周抗体水平即高于1∶800,达到猪圆环病毒2型灭活疫苗的国家质量标准要求,3周时达到高峰,比对照商品疫苗诱导产生的抗体水平更高;其分别稀释130倍和463倍后制备的疫苗诱导产生的抗体水平在二免后3周也能达到1∶800。提示,猪圆环病毒2型8#和12#分离株毒价高,免疫效力良好,是理想的疫苗候选株。本研究丰富了猪圆环病毒2型灭活疫苗毒株的种毒资源,为研制高质量的猪圆环病毒2型疫苗奠定了基础。 相似文献
5.
用水包油佐剂Merckinade SDA 25、HS1010和双向佐剂Montanide ISA 206三种不同佐剂分别制成猪圆环病毒2型灭活疫苗，对三种疫苗的稳定性、黏度、免疫仔猪后的安全性及效力进行了比较。试验结果表明，以3 000 r/min离心15 min，三种疫苗均无水析出；三种疫苗的黏度均低于200 cP；三种疫苗按2.0 mL/头的剂量接种21日龄健康易感猪，试验猪体温、采食、饮水、精神状况均正常，均无不良临床反应，剖检后各脏器均无病理变化；加强免疫后21d，各组试验猪采血，检测猪圆环病毒2型抗体效价，结果表明，佐剂MontanideISA206相较MerckinadeSDA25可诱导更高水平抗体效价。攻毒试验结果表明免疫组可以得到100%的保护。根据以上试验结果，确定猪圆环病毒2型灭活疫苗使用Montanide ISA 206。 相似文献
6.
《中国预防兽医学报》2020,(6)
为比较3种TLR配体对猪圆环病毒2型(PCV2)病毒样颗粒(VLP)疫苗的佐剂活性,本研究利用重组大肠杆菌表达脑膜炎奈瑟菌Ag473脂蛋白、创伤弧菌FlaB鞭毛蛋白和结核分枝杆菌热应激蛋白70(Hsp70),采用镍亲和柱纯化获得该3种重组蛋白;分别将相同剂量(10μg)3种重组蛋白、ISA206佐剂(100μL)和不完全弗氏佐剂(FIA)(100μL)与PCV2 VLP疫苗(50μg)混合,肌肉注射免疫小鼠,初免后14 d加强免疫1次;初免后每周采血分离血清,采用ELISA检测抗原特异抗体;加强免疫后14 d利用PCV2攻毒,攻毒后7 d和14 d采血分离血清,采用荧光定量PCR检测病毒DNA拷贝数;攻毒后14 d迫杀小鼠,制备并培养其脾细胞,经PCV2 VLP疫苗刺激后,采用试剂盒检测细胞因子的表达。SDS-PAGE分析结果显示,3种TLR配体在重组大肠杆菌中均获得正确表达,纯化重组蛋白的纯度大于90%。从初免14 d起,3个分子佐剂组中Ag473佐剂组的PCV2抗体水平最高,但低于ISA206佐剂组却高于FIA组,差异不显著(p0.05)。在PCV2攻毒前,3个分子佐剂组中Ag473佐剂组的小鼠脾细胞TNF-α和IFN-γ表达水平最高,但低于ISA206佐剂组却高于FIA组,差异显著(p0.05);Hsp70佐剂组的IL-4表达水平最高,Ag473佐剂组次之,但均高于ISA206和FIA组。在攻毒后第7 d,5个免疫组小鼠的病毒拷贝数均较对照组显著下降(p0.05),但免疫组间差异不显著;在攻毒后第14 d,5个免疫组小鼠的病毒拷贝数进一步下降,免疫组间差异不显著。研究结果表明在测试的3种分子佐剂中,Ag473脂蛋白具有较强的总体免疫佐剂活性,有望进一步开发为PCV2等病毒的亚单位疫苗佐剂。 相似文献
7.
《畜牧与兽医》2020,(8)
试验采用阻断ELISA试剂盒和间接ELISA试剂盒,分别对人工感染猪和自然感染猪进行了猪圆环病毒2型(porcine circovirus 2, PCV2)抗体检测。结果显示:PCV2攻毒组和PCV2攻毒后佐剂KHL刺激组试验猪用两种ELISA试剂盒检测,检测结果基本一致。PCV2抗体均于攻毒后2～3周出现,6～8周达到最高,至第16周仍维持较高水平。PCV2攻毒后佐剂刺激组猪产生的抗体水平明显高于PCV2攻毒组。PCV2自然感染猪用两种ELISA试剂盒检测,检测结果一致。断奶仔猪多系统衰竭综合征(PMWS)发病猪抗体阳性率为99%,PMWS亚临床感染猪抗体阳性率为95%,PMWS发病猪抗体水平多数高于亚临床感染猪。 相似文献
8.
为了探讨猪繁殖与呼吸综合征病毒(PRRSV)N蛋白基因工程亚单位疫苗的免疫保护效果,试验利用大肠杆菌表达并纯化了重组PRRSV N蛋白,添加佐剂制成亚单位疫苗进行免疫保护试验。试验猪分为2个不同佐剂制备的PRRSV N蛋白亚单位疫苗免疫组,1个PRRSV TJM-F92株减毒活疫苗免疫组和1个攻毒对照组。首免5周后,用PRRSV NVDC-JXA1强毒株攻击,观察临床表现并在攻毒后21 d对试验猪进行剖检。结果表明:首免3周后2个亚单位疫苗组和减毒活疫苗组抗体全部转阳,组间无显著性差异(P0.05);攻毒后PRRSV N蛋白亚单位疫苗免疫1#组有2/3的猪只、2#组有1/3的猪只的临床症状、体征和解剖病变较攻毒对照组猪只轻微,转归较好且成活,但保护作用不及PRRSV TJM-F92株减毒活疫苗组;剩余的亚单位疫苗免疫1#组1/3的猪只和亚单位疫苗免疫2#组2/3的猪只,与攻毒对照组猪只一样表现为比较典型的PRRS症状,并在8~12 d内死亡。说明PRRSV N蛋白亚单位疫苗能够引起机体的体液免疫反应,具有部分保护作用,但不能提供完全的保护,单独使用达不到减毒活疫苗的效果。 相似文献
9.
10.
PCV2a/2b灭活疫苗与PCV2b亚单位疫苗免疫效力比较试验 总被引：1，自引：0，他引：1
比较猪圆环病毒(PCV)2a、PCV2b基因型全病毒灭活疫苗和PCV2b亚单位疫苗对PCV2b基因型强毒株免疫攻毒保护效力。取3种类型PCV2疫苗,分2次免疫PCV2抗原和特异性抗体阴性的BALB/c小鼠和14日龄仔猪,间隔21d,分别于免疫后14、21和35 d进行PCV2特异性ELISA抗体测定。使用临床分离鉴定的PCV2b基因型强毒株进行攻毒,通过攻毒前后临床观察、体温变化、平均相对日增重及PCV2核酸载量检测等对疫苗免疫效果进行评价。结果:不同类型PCV2疫苗免疫BALB/c小鼠后14 d能检测到PCV2特异性抗体,在35 d时抗体水平持续升高,其中PCV2b亚单位疫苗产生抗体水平高于全病毒灭活疫苗;不同类型疫苗均能刺激仔猪产生较好的免疫应答,其中PCV2b亚单位疫苗刺激机体免疫应答强于灭活疫苗;仔猪免疫攻毒试验表明,3种类型疫苗均可刺激机体产生抵御PCV2强毒攻击的特异性免疫应答,免疫组体温变化、相对日增重及PCV2核酸载量检测均与攻毒对照组间存在显著性差异,3种类型疫苗之间差异不显著。试验表明:3种类型PCV2疫苗免疫小鼠和仔猪后均能诱导产生抵御PCV2b基因型强毒攻击的特异性免疫应答反应,有效抵抗PCV2感染和提高猪的生长性能。 相似文献
11.
以猪圆环病毒2型(PCV2)遗传标记毒株为毒种,经细胞培养传代,优化了病毒增殖条件,获得较高滴度的病毒培养物用于PCV2灭活疫苗的研制。为了比较不同免疫佐剂的效果,本试验对国产矿物质白油和铝胶两种佐剂以及赛比克公司提供的MONTANIDETMISA206、ISA15VG、IMS1315VG3种佐剂配制的灭活疫苗进行了猪体免疫试验。通过临床观察、血清抗体效价测定,确认ISA15VG佐剂配制的疫苗在增强免疫效果方面优于其它佐剂。按5种佐剂免疫效果排序为ISA15VG、IMS1315VG、铝胶、ISA206、国产白油。ISA15VG佐剂属于水包油剂型,可刺激接种动物产生快速免疫应答反应,抗体产生效价高、持续时间长,有可能成为新型PCV2灭活疫苗佐剂的候选。 相似文献
12.
The study was aimed to prepare vaccines with different adjuvants,and research its effects on immunogenicity.The PCV2 Cap gene with its signal peptide removed was connected to pET-28a vector,and then was induced to express,using sodium deoxycholate（DOC）and low concentration of urea to dissolve the inclusion body.Different adjuvants,such as alum-based adjuvants,liposome adjuvants,propolis adjuvants,white oil adjuvants and Freund adjuvant were prepared,together with protein purified to make up subunit vaccines,and commercial inactivated vaccine as positive control,immuning mice,and ELISA method were used to detect changes in concentrations of animal serum antibody and cytokines,evaluated the immune protective effect.Results showed that the expression product in the form of inclusion body,using the DOC could omit dissolving inclusion body protein renaturation steps,and the purification method was simple,the capsid protein obtained had high purity.ELISA assays showed that PCV2-Cap had good immunogenicity.And we found that the water system adjuvants had high immune activity,this could provide important previous experimental data for the commercialization of the subunit vaccine. 相似文献
13.
试验旨在研究猪圆环病毒不同佐剂疫苗的制备及其对免疫原性的影响。将去除信号肽的PCV2（porcine circovirus type 2,PCV2）Cap蛋白基因连接在pET-28a载体上,进行诱导表达,用脱氧胆酸钠（DOC）和低浓度尿素对表达产物进行溶解,制备氢氧化铝胶体佐剂、脂质体佐剂、弗氏佐剂、白油佐剂、蜂胶佐剂,并与纯化的PCV2-Cap蛋白混合制成亚单位疫苗,以商品化的灭活疫苗作为阳性对照,免疫小鼠,并使用ELISA方法检测动物血清中抗体及细胞因子含量的变化,评估其免疫保护效果。结果显示,表达产物以包涵体的形式存在,使用DOC溶解包涵体可省略蛋白复性步骤,且纯化方法简单,获得纯度较高的衣壳蛋白;ELISA检测结果表明PCV2-Cap蛋白能诱导产生特异性较高的抗体;水系佐剂制备的亚单位疫苗具有较高的免疫活性,为亚单位疫苗的商品化提供重要的数据支持。 相似文献
14.
猪圆环病毒2型(PCV2)感染后会导致断奶仔猪多系统衰竭综合征(PMWS)等"猪圆环病毒相关疾病"(PCVADs),给全球养猪业造成了巨大的经济损失。疫苗免疫是防控该病的有效手段,近年来市场上推出了PCV2灭活疫苗、嵌合疫苗和亚单位疫苗。PCV2变异较快,主要临床流行毒株从PCV2b逐渐演变为PCV2d,现有商业化疫苗免疫效果需进一步研究证实。论文就近年来国内外PCV2传统疫苗、新型基因工程疫苗、PCV2疫苗与其他疫苗联合免疫的研究进展进行综述,旨在为猪圆环病毒相关疾病的防控及疫苗的开发与利用提供参考。 相似文献
15.
The objective of this study was to evaluate the effect of porcine circovirus type 2 (PCV2) vaccines on PCV2-viremic and -seropositive piglets born from naturally PCV2-infected sows against postnatal PCV2 challenge. The experimental design was aimed at mimicking commercial swine rearing conditions to evaluate the response of the PCV2 vaccine on PCV2-viremic and -seropositive piglets after experimental PCV2 challenge. PCV2a (or 2b)-viremic piglets received a PCV2 vaccine at 21 days of age followed by a PCV2b (or 2a) challenge at 49 days of age (28 days post vaccination). The PCV2 vaccines elicited a high level of humoral (as measured by immunoperoxidase monolayer assay and neutralizing antibody titers) and cellular (as measured by the frequency of PCV2-specific interferon-γ-secreting cells) immune response in the PCV2-viremic piglets after vaccination even in the presence of maternally derived antibodies (MDA). The initial infection of PCV2 in the pigs was not affected by PCV2 vaccination, however the challenging PCV2 was reduced by PCV2 vaccination on PCV2-viremic pigs. The results from this study demonstrate that the PCV2 vaccine used in this study is effective at reducing PCV2 viremia and lymphoid PCV2 DNA, even for PCV2-viremic pigs with passively acquired MDA at the time of vaccination. 相似文献
16.
为了解免疫不同圆环病毒病疫苗的抗体水平,给养殖户提供合适的免疫依据,对福建省不同地区8个猪场在使用不同猪圆环病毒病疫苗和免疫程序后猪群的抗体水平进行检测。结果显示:免疫场和未免疫场各阶段种猪群的猪圆环病毒病抗体阳性率皆为100%,差异不显著(P>0.05);但各阶段种猪群的S/P平均值,免疫场较未免疫场更高;同时通过比较变异系数发现,未免疫场种猪的抗体水平更活跃;14日龄免疫亚单位疫苗后抗体水平较高且维持时间较长,而免疫全病毒灭活苗后抗体水平仍持续下降。综上,种猪群免疫圆环病毒病疫苗是必要的,亚单位疫苗比全病毒灭活苗的免疫效果好。 相似文献
17.
A field evaluation of mortality rate and growth performance in pigs vaccinated against porcine circovirus type 2 总被引：3，自引：0，他引：3
Horlen KP Dritz SS Nietfeld JC Henry SC Hesse RA Oberst R Hays M Anderson J Rowland RR 《Journal of the American Veterinary Medical Association》2008,232(6):906-912
OBJECTIVE: To evaluate, under field conditions, the effects of a commercial porcine circovirus type 2 (PCV2) vaccine on mortality rate and growth performance in a herd infected with PCV2 that had a history of porcine circovirus disease. DESIGN: Randomized controlled clinical trial. ANIMALS: 485 commercial, cross-bred, growing pigs. PROCEDURES: Prior to weaning, pigs were randomly assigned within litter to a vaccination or unvaccinated control group. Pigs in the vaccination group were given a commercial PCV2 vaccine at weaning and 3 weeks later. Mortality rate was recorded, and pigs were weighed prior to vaccination, when moved from the nursery, and prior to marketing. Infection status was assessed by serologic testing and detection of viral DNA in serum. RESULTS: Compared with control pigs, pigs vaccinated against PCV2 had a significantly lower mortality rate during the finishing phase, significantly higher average daily gain during the finishing phase, and significantly lower likelihood of being lightweight at the time of marketing. For vaccinated pigs, overall mortality rate was reduced by 50% and average daily gain during the finishing period was increased by 9.3%. At the time of marketing, vaccinated pigs weighed an average of 8.8 kg (19.4 lb) more than control pigs, without any difference in days to marketing. Serum PCV2 antibody titers increased in control pigs, and PCV2 DNA was detected, indicating active PCV2 infection. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that vaccination against PCV2 was effective at reducing mortality rate and improving growth performance among pigs in a herd infected with PCV2. 相似文献
18.
不同种类的佐剂对同种抗原的免疫辅佐效力不同。为筛选绵羊肺炎支原体(Mycoplasma ovipneumoniae,MO)灭活疫苗的理想佐剂,本研究制备了4种不同佐剂的MO灭活疫苗,分别免疫成年家兔,用MO间接血凝试剂盒测定免疫后1～9周血清抗体效价。检测结果显示,4种疫苗在免疫后都能迅速产生抗体,其中用MO Buonavoglial's和ISA-760佐剂疫苗免疫家兔,产生抗体均于免疫后第3周达到高峰((5.67±0.70) log2,6.00 log2),到第9周时维持抗体高峰(6.00 log2,(6.00±1.00) log2);MO ISA-206和白油佐剂疫苗免疫抗体均于免疫后第4周达到高峰((4.33±0.57) log2,5.00 log2),至第9周下降约1个滴度。统计学分析结果显示,ISA-760、Buonavoglial's佐剂对MO的免疫增强效力显著优于ISA-206和白油佐剂(P<0.05),是MO灭活疫苗的理想候选佐剂,为高效MO灭活疫苗的研发提供了科学依据。 相似文献
19.
为研究比较5种塞内卡病毒疫苗佐剂,分别用4种自主研发佐剂和进口ISA206佐剂配制塞内卡病毒灭活疫苗,检测疫苗的理化性质、安全性及免疫效力,并对检测结果进行对比分析。结果显示,进口ISA206佐剂疫苗黏度为43.02 cP,自主研发佐剂疫苗黏度均在27.88 cP以下,其他理化性质无差异;5批疫苗安全性试验均未见明显异常;5批疫苗免疫动物后均可诱导机体产生中和抗体,二免后14 d抗体滴度在211以上,4批自主研发佐剂疫苗免疫组抗体滴度水平高于进口ISA206佐剂疫苗约1个滴度,攻毒保护结果均在4/5以上,2批自主研发佐剂疫苗免疫组保护率可达5/5。结果表明,用4种自主研发佐剂制备的塞内卡病毒灭活疫苗质量不低于用进口ISA206佐剂制备的塞内卡病毒灭活疫苗。 相似文献
20.
Danyang Wang Yu Yang Jinyu Li Bin Wang Ailian Zhang 《Journal of veterinary science (Suw?n-si, Korea)》2021,22(3)
BackgroundNew-generation adjuvants for foot-and-mouth disease virus (FMDV) vaccines can improve the efficacy of existing vaccines. Chinese medicinal herb polysaccharide possesses better promoting effects.ObjectivesIn this study, the aqueous extract from Artemisia rupestris L. (AEAR), an immunoregulatory crude polysaccharide, was utilized as the adjuvant of inactivated FMDV vaccine to explore their immune regulation roles.MethodsThe mice in each group were subcutaneously injected with different vaccine formulations containing inactivated FMDV antigen adjuvanted with three doses (low, medium, and high) of AEAR or AEAR with ISA-206 adjuvant for 2 times respectively in 1 and 14 days. The variations of antibody level, lymphocyte count, and cytokine secretion in 14 to 42 days after first vaccination were monitored. Then cytotoxic T lymphocyte (CTL) response and antibody duration were measured after the second vaccination.ResultsAEAR significantly induced FMDV-specific antibody titers and lymphocyte activation. AEAR at a medium dose stimulated Th1/Th2-type response through interleukin-4 and interferon-γ secreted by CD4+ T cells. Effective T lymphocyte counts were significantly elevated by AEAR. Importantly, the efficient CTL response was remarkably provoked by AEAR. Furthermore, AEAR at a low dose and ISA-206 adjuvant also synergistically promoted immune responses more significantly in immunized mice than those injected with only ISA-206 adjuvant and the stable antibody duration without body weight loss was 6 months.ConclusionsThese findings suggested that AEAR had potential utility as a polysaccharide adjuvant for FMDV vaccines. 相似文献