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1.
将转化C型节瘤拟杆菌纤毛蛋白基因表达子粒的工程菌PAK/2pfsC在含羧苄青霉素营养肉汤培养基中培养,用MgCl2法在培养物上清中提取表达产物(纤毛蛋白).将表达的纤毛蛋白产物用弗氏完全佐剂乳化,制成疫苗.用疫苗免疫3只健康家兔,21 d后再次接种;定期采血,用对流免疫电泳和K凝集实验检测试验家兔的体液免疫应答及抗体水平.结果发现,免疫7 d即可产生相应抗体,21 d后抗体效价达到8 000×以上,而且高滴度抗体可维持6个月以上.试验表明,腐蹄病C型节瘤拟杆菌纤毛蛋白基因工程疫苗具有较好的免疫应答和免疫原性.  相似文献   

2.
坏死梭杆菌毒力菌株FN(AB)94免疫原对鹿的初步免疫试验   总被引:1,自引:1,他引:0  
本研究将坏死梭杆菌毒力菌株FN(AB)94厌氧培养后,裂解制备抗原,与等量福氏完全佐剂混合,制备乳化佐剂胺苗。将疫苗分别接种4头不同年龄的健康成年鹿(每头接种4ml)。3头鹿作为对照组。接种前,静脉采血,进行对流免疫电泳检查。30天后,分别用150-400亿菌感染试验动物,然后混群饲养观察,并定期采血检查被免鹿血清中抗体消长变化。结果对照组鹿分别于3个月后开始现现明显的临床症状,而免疫组不表现任何临床症状,6个月仍表现临床健康。4头免疫鹿6个月时都能检测到血清抗体,3头免疫鹿血清抗体可维持到7个月后。试验表明。试验表明,坏死梭杆菌毒力菌株FN(AB)94疫苗接种鹿后,可刺激鹿产生很好的免疫力,从而通过本动物初步证明坏死梭杆菌毒力菌株FN(AB)94免疫原具有良好的免疫原性,可以用于制备坏死杆菌病疫苗。  相似文献   

3.
用牦牛大肠埃希氏菌灭活疫苗对实验动物家兔进行疫苗稳定性(保存期)测定。4批牦牛大肠埃希氏菌病灭活疫苗,批号分别为200401、200501、200502、200503,将这4批疫苗4℃保存3、6、9、12个月后,分别接种试验兔,每批接种5只。初次接种疫苗0.5mL/只,14d再次接种疫苗1.0mL/只。每次设对照组5只,注射肉汤,其剂量和方法与试验兔相同。疫苗接种前和再次接种疫苗后21d分别在试验和对照家兔耳静脉采血,用微量凝集试验进行抗体效价检测。通过抗体检测表明,疫苗保存了3、6、9、12个月后接种兔,采血测免疫抗体效价为2.709~3.612。同时对照组全部死亡。证明这些疫苗存放12个月能有效保护家兔免受大肠杆菌攻击。  相似文献   

4.
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   

5.
为了筛选绵羊肺炎支原体(MO)灭活疫苗的佐剂,试验采用绵羊肺炎支原体新疆分离株扩大培养后得到灭活抗原,分别与两种不同佐剂ISA 201和ISA 206混合乳化制备灭活疫苗,再以不同剂量免疫健康成年小鼠,最后用MO间接血凝的方法分别测定二免后第7,14,21,28天血清抗体效价。结果表明:ISA 206佐剂疫苗0.3 m L剂量组在第14天抗体效价就达到较高水平,第21天后开始下降;ISA 206佐剂疫苗0.2 m L剂量组在二免后第14天抗体效价达到高峰(1∶128),且在第21天仍维持抗体高峰,第28天开始下降;ISA 201佐剂疫苗0.3 m L剂量组在二免疫后第14天抗体效价达到高峰,维持1周后开始下降;ISA 201佐剂疫苗0.2 m L组免疫应答较慢,在二免后第21天才达到最高抗体效价(1∶64)。说明ISA 206佐剂诱导产生MO抗体较快,维持时间长,优于ISA 201佐剂,可作为MO灭活疫苗的优选佐剂。  相似文献   

6.
在注射兔病毒性出血症 (RHD)疫苗后 ,30只兔随机分为 2组 ,试验组兔饲喂含 1 5 %黄白散的饲料。从接种到第 1 80天 ,每组取兔 8只 ,每隔 1 5d采血 1次 ,检测血凝抑制 (HI)抗体 ;在接种前 1天及接种后 5、 1 0、 1 7、 2 4、 31、 41、 5 1d ,每组取兔 1 0只 ,采血检测ANAE+淋巴细胞百分率 ;在 1 80、 2 4 0、 30 0d时 ,每组分别取兔 5只做攻毒保护试验。结果表明 :2组兔HI抗体峰值分别为 7 3log2和 9 8log2 ,差异极显著 (P <0 0 1 ) ;试验组疫苗保护期可延长 60d ;试验组ANAE+率从第 1 0天至 5 1天与对照组比较 ,差异极显著(P<0 0 1 )。所以黄白散可增强兔接种RHD疫苗后的特异性免疫功能 ,增强RHD疫苗的免疫效果。  相似文献   

7.
不同佐剂对鸡新城疫疫苗免疫调节作用的影响   总被引:1,自引:0,他引:1  
试验以300只AA雏鸡为试验动物,将其随机分为4组,A、B、C组分别接种相同剂量的甲壳素、蜂胶、铝胶佐剂苗,D组为对照组。结果表明,三种佐剂的疫苗在对肉鸡组免疫的第3、4、5、6、7、9、11、13、15、20、25天后,甲壳素疫苗组H I抗体效价显著高于其它组,蜂胶疫苗组H I抗体效价其次,铝胶疫苗组抗体H I效价最低;用三种不同佐剂的疫苗对肉鸡免疫后第26天称其重量,甲壳素佐剂疫苗组免疫后的肉鸡增重效果最好,蜂胶疫苗组居中,铝胶疫苗组最差。  相似文献   

8.
猪囊虫病基因工程疫苗的体液与细胞免疫反应   总被引:2,自引:0,他引:2  
为了探讨以 I S C O M 作佐剂的猪囊虫病基因工程疫苗的免疫机理,对其诱导的体液免疫与细胞免疫反应进行了测定。用上述疫苗免疫 9 头试验猪,采用间接 E L I S A 检测体液免疫反应及通过淋巴细胞转化试验、 A N A E染色试验、 E玫瑰花环形成试验等检测细胞免疫反应;用该疫苗和铝胶苗分别免疫昆明小鼠各 20 只,分别检测体液免疫反应和 T 淋巴细胞抑制/杀伤亚群的动态变化。体液免疫的检测结果显示,免疫后 7 天即出现抗体,21 天后抗体全部转阳,持续的时间不少于 193 天,效价明显高于铝胶苗;细胞免疫检测结果显示,免疫猪外周血 T淋巴细胞转化率、 A N A E+ 细胞和粗粒型 A N A E+ 细胞、 E R F C和 Ea R F C细胞显著升高,免疫小鼠 T淋巴细胞抑制/杀伤亚群显著升高;与铝胶苗及对照组比较,差异极显著。以上结果表明猪囊虫病基因工程疫苗可同时激发动物的体液免疫和细胞免疫反应,增强了机体的免疫调节功能及杀伤性 T 淋巴细胞功能。  相似文献   

9.
筛选了4株氨基酸序列差异显著的超强毒力传染性法氏囊病毒(vvIBDV)的细胞弱化毒株GXCL1-25、GXCL2-30、GXCL4-25、GXCL5-30制备活疫苗(加入泰山松花粉多糖为免疫佐剂,蜂胶佐剂作为对照),并分别对这些疫苗的免疫效果进行分析。分别测定4株病毒克隆株的TCID50,并分别制备相应的佐剂活疫苗进行接种试验。对4株病毒克隆分别设置无免疫佐剂疫苗接种组(A1、B1、C1、D1)、松花粉多糖佐剂疫苗接种组(A2、B2、C2、D2)、蜂胶佐剂疫苗接种组(A3、B3、C3、D3)及空白对照组(E)。每组接种SPF鸡50只,于14日龄首免,25日龄二免。分别于一免后0、3、7d和二免后3、10、17、24d采血,用ELISA方法检测血清抗体效价和IL-2分泌水平;28日龄与42日龄测定外周血淋巴细胞比率,28日龄每组剖杀5只测免疫器官指数;35日龄进行攻毒试验。结果显示,各试验组的抗体效价和IL-2水平均在35日龄时(二免后第10天)达到最高峰,但佐剂疫苗组与无佐剂疫苗组相比具有更高的IBDV抗体水平、更长的抗体维持时间长、更轻的法氏囊损伤,并且攻毒保护率均在90%以上;尤其松花粉多糖佐剂疫苗组的抗体效价、IL-2水平和淋巴细胞比率比其他组更高,且松花粉多糖为佐剂的GXCL4-25组免疫效果最好。结果表明,以泰山松花粉多糖为佐剂的IBDV细胞弱化活疫苗表现出良好的免疫原性和保护力。  相似文献   

10.
用猪瘟毒弱毒株感染或活毒疫苗接种猪,探讨了体液免疫反应(血清中和作用)和细胞免疫反应(淋巴细胞刺激试验)。接种疫苗的猪显现出强烈地产生中和抗体,但没有查出细胞免疫反应。而在感染慢性猪瘟病毒的猪中,未发现中和抗体,但观察到暂短时间的细胞免疫反应(在接种后18天)。  相似文献   

11.
Inoculation of calves with potassium thiocyanate (KSCN) extract of Pasteurella multocida type A in saline-tris buffer or in Freund's incomplete adjuvant or modified Freund's incomplete adjuvant resulted in the elicitation of agglutinating, hemagglutinating, bactericidal and homocytotropic antibodies. The antibody response was significantly (P = 0.05) higher in calves inoculated with the KSCN extract in either adjuvant than in those inoculated with the extract in saline-tris buffer. Hemagglutination also was observed if the KSCN extract of P hemolytica was used for sensitizing the tanned sheep red blood cells. Further, calf anti-P multocida extract antisera also was bactericidal to P hemolytica. The KSCN extract of P multocida was found to be nontoxic to calves at 2 doses tested, as judged by an evaluation of total and differential leukocyte counts, body temperature, and pulse rates at various intervals after inoculation.  相似文献   

12.
Immunoaffinity-purified bovine respiratory syncytial virus (BRSV) fusion (F) protein elicited anti-BRSV-specific antibody responses in BRSV-seronegative calves. After primary vaccination, all calves seroconverted to BRSV as determined by the virus neutralization (VN) test and developed anti-F protein antibodies detectable by protein immunoblot analyses. Subsequent vaccinations induced greater than twofold increase in VN titer in 3 of 9 (33%) calves, and 1 calf became VN-negative, but still had nonneutralizing antibody detectable by protein immunoblot analysis. This calf remained seronegative after challenge exposure. Two groups of calves were vaccinated IM with immunoaffinity-purified BRSV F protein. Each dose was 2 ml containing 20 micrograms of purified F protein. Freund's adjuvants were used for all vaccinations, with Freund's complete adjuvant used for the primary vaccination and Freund's incomplete adjuvant for subsequent vaccinations. The vaccine was administered to both groups at weeks 0 and 3; the first group received a third vaccination at weeks 21. Group-1 and -2 vaccinated calves and non-vaccinated contact controls were intranasally aerosol challenge-exposed with low cell culture-passage BRSV on weeks 22 and 9, respectively. Eight of 9 vaccinated calves did not develop a humoral anamnestic response following challenge exposure, as demonstrated by VN test and protein immunoblot analyses. Calf 14 from group 1 which had a 1:2 VN antibody titer prior to vaccination, was the only calf that developed an anamnestic response.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

13.
Group-specific antibodies were produced by inoculation of bluetongue virus soluble antigen into polyethylene chambers implanted subcutaneously in 8 rabbits and 2 sheep. For comparison, 5 rabbits and 1 sheep were inoculated intramuscularly with the soluble antigen in Freund's complete adjuvant. Antibodies present in the serum and chamber fluids were detected by the agar gel precipitin or serum-neutralization tests, qualitatively examined by immunoelectrophoresis and immunofluorescence, and quantitated by electroimmunodiffusion.  相似文献   

14.
Merino sheep vaccinated with either whole Bacteroides nodosus organisms, a crude surface antigen preparation or highly purified pili (>99% homogeneity) in oil adjuvant, developed significant resistance to artificial footrot infection when compared with unvaccinated control sheep inoculated with saline-in-oil emulsion (Freund;s incomplete adjuvant) alone. The pili-vaccinated sheep generally had higher K-agglutinating antibody titres than sheep vaccinated with whole B. nodosus. These results confirmed the role of B. nodosus pilus protein both as a protective antigen and the K-agglutinogen. Vaccines prepared with Freund;s incomplete adjuvant containing either purified pili, crude pili or B. nodosus whole cells did not produce significantly different injection-site reactions.  相似文献   

15.
Sodium diethyldithiocarbamate (DTC), mycobacterium cell wall extract (MCWE, Regressin), killed Corynebacterium parvum (C. parvum, Immunoregulin) and muramyldipeptide (MDP) were each combined with purified, live bovine rotavirus and inoculated into 3 month-old Holstein-Friesian calves in order to examine their ability to potentiate specific humoral and cellular immune responses. The vaccinated calves were boosted twice at 3 and 6 weeks after initial vaccine inoculation. The rotavirus was administered intramuscularly either in an aqueous suspension or in a water-in-oil (WIO) emulsion, prepared with incomplete Freund's adjuvant (IFA). DTC and C. parvum were given by the intravenous route, while MCWE and MDP were incorporated directly in the rotavirus suspension. Two groups of calves were also vaccinated either with rotavirus and IFA or with rotavirus emulsified in mineral oil and a mannide oleate compound (MOC, Montanide 888). A control group of calves was given phosphate-buffered saline (PBS) solution emulsified with IFA. The different vaccine preparations were then compared by studying the kinetics of serum rotavirus-neutralizing antibody production and of proliferative response by blood lymphocytes following in vitro stimulation with bovine rotavirus. The results showed that: (1) the bovine rotavirus should be incorporated in a WIO emulsion in order to induce a cell-mediated immune response as detected by the rotavirus-specific in vitro stimulation test with blood lymphocytes, and to produce higher neutralizing antibody titers in the serum; (2) the vaccines prepared with the mineral oil-MOC complex or IFA both induced comparable levels of humoral and cellular immune responses. The use of mineral oil and MOC as adjuvant may be preferred to IFA, because of the facility of preparing the vaccine and of the low viscosity of the resulting WIO emulsion: (3) the addition of MDP to the WIO emulsion prepared with IFA resulted in a higher cell-mediated immune response as determined by the in vitro blood lymphocyte transformation index specific for bovine rotavirus.  相似文献   

16.
Merino sheep vaccinated with either whole Bucteroides nodosus organisms, a crude surface antigen preparation or highly purified pili (>99% homogeneity) in oil adjuvant, developed significant resistance to artificial footrot infection when compared with unvaccinated control sheep inoculated with saline-in-oil emulsion (Freund's incomplete adjuvant) alone. The pili-vaccinated sheep generally had higher K-agglutinating antibody titres than sheep vaccinated with whole B. nodosus. These results confirmed the role of B. nodosus pilus protein both as a protective antigen and the K-agglutinogen.

Vaccines prepared with Freund's incomplete adjuvant containing either purified pili, crude pili or B. nodosus whole cells did not produce significantly different injection-site reactions.  相似文献   

17.
Objective To compare the effects of two oil emulsion adjuvants (incomplete Freunds adjuvant and a proprietary oil adjuvant), DEAE-dextran, L-tyrosine particles and Quil A on the humoral immune responses of sheep immunised with recombinant pili of Dichelobacter nodosus (strain A).
Procedure Antibody titres were studied for up to 32 weeks and were measured by bacterial agglutination and ELISA. The relative avidity of antibodies for pili was determined and the incidence and severity of adverse reactions at the site of injection of vaccines were recorded.
Results The oil emulsion adjuvants and Quil A were more effective than either DEAE-dextran or L-tyrosine at stimulating antibodies in sheep. The incidence and severity of adverse reactions was lower in sheep which received vaccines containing either Quil A or DEAE-dextran than in sheep which received vaccines containing oil emulsion adjuvants. L-tyrosine had no adverse effects.
Conclusion Quil A was as effective as oil adjuvants at stimulating high levels of antibodies against recombinant pili in sheep and had the significant advantage of being less irritant after subcutaneous injection.  相似文献   

18.
Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis, a disease that affects goats and sheep, and can cause severe economic losses. In this study, four different antigenic extracts were obtained from the attenuated strain T1, which was isolated in the state of Bahia (Brazil). Forty-four Canindé breed goats were divided in five groups, each receiving a different antigen solution and saline buffer as a control. The humoral response was monitored through the identification of specific IgG by indirect ELISA and Western Blotting, and the production of IFN-gamma was followed in order to observe the activation of cellular response. After twelve weeks of antigen inoculation, the animals were challenged with 2 x 10(5)CFU of a wild strain, also isolated in Bahia, and necropsy was performed on all animals twelve weeks afterwards. It was observed that the attenuated bacteria gave a protection of 33.3%, in addition to the weak humoral response elicited. Animals inoculated with secreted antigen associated with Freund's incomplete adjuvant and oligodeoxynucleotide containing unmethylated CpG dinucleotides (CpG ODN) showed a strong humoral response, but this inoculation could not prevent the spread of challenge bacteria in the majority of animals. These results demonstrate the immunogenic potential of the attenuated T1 strain in the development of a vaccine against caseous lymphadenitis in goats.  相似文献   

19.
Groups of sheep were immunised twice with one or other of six vaccines consisting of purified pili from Bacteroides nodosus at three dose levels (10, 38 and 154 micrograms) and emulsified with either complete (CFA) or incomplete Freund's adjuvant (IFA). Beginning one month after vaccination the sheep were homologously challenged on irrigated pasture, with naturally transmitted foot rot for a period of 26 weeks. Statistical analyses of the number of feet per sheep with severe foot rot demonstrated that there was a significant effect of vaccinal dose but neither an adjuvant effect nor an interaction between dose and adjuvant. Similar conclusions were reached when the titres of antipilus agglutinins in the serum were analysed. By both criteria the responses to doses of 154 and 38 micrograms of pili were significantly better than to 10 micrograms, but not significantly different from each other. The IFA vaccines caused less reaction at the sites of injection than the CFA vaccines and within the former the vaccines containing 10 and 38 micrograms pilus produced less reaction than those containing 154 micrograms. Hence a vaccine containing 38 micrograms of purified pili in IFA is nearly optimal for homologous protection against severe foot rot and is acceptable in terms of the reaction at the injection site.  相似文献   

20.
白细胞介素-2对犬细小病毒疫苗免疫效果的影响   总被引:3,自引:2,他引:1  
为研究白细胞介素-2(IL-2)对犬细小病毒疫苗免疫效果的影响,选择40只幼犬,随机分成试验组和对照组,试验组按0.5 ml/头份加入犬用IL-2与犬细小病毒疫苗一起皮下注射,对照组单独使用犬细小病毒疫苗,注射后分别在接种前、接种后7 d和14 d使用金标试纸条检测其抗体效价,并进行组间抗体效价差异的t检验。试验结果表明,接种疫苗前试验组与对照组犬细小病毒平均抗体效价之间差异不显著;接种疫苗后7 d,试验组与对照组犬细小病毒平均抗体效价差异极显著(P<0.01);接种疫苗后14 d,试验组与对照组犬细小病毒平均抗体效价差异极显著(P<0.01)。试验表明:IL-2可明显提高幼犬对犬细小病毒疫苗的抗体应答能力,是一种良好的犬细小病毒疫苗候选佐剂。  相似文献   

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