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1.
Fusion expression of major antigenic segment of JEV E protein-hsp70 and the identification of domain acting as adjuvant in hsp70 总被引：3，自引：0，他引：3
Ge FF Qiu YF Gao XF Yang YW Chen PY 《Veterinary immunology and immunopathology》2006,113(3-4):288-296
Hsp70 potentiates specific immune responses to some antigenic peptides fused to it. A recombinant hsp70 protein expression vector in methylotrophic yeast, Pichia pastoris, was developed that fused the major antigenic segment of Japanese encephalitis virus (JEV) E protein to the amino terminus of Mycobacterium tuberculosis hsp70. The C-terminal peptide binding domain of hsp70 stimulated Th1-polarizing cytokines, CC chemokines and an adjuvant effect. However, the N-terminal ATPase domain (hsp70 1-358) failed to stimulate any of these cytokines or chemokines. Based on these data, a vector was constructed that permits the fusion of major antigenic segment of E protein to the amino terminus of peptide binding domain of hsp70. Antibody titers, lymphocytes proliferation, the level of mIL-2 or mIFN-gamma and neutralizing antibodies in immunized mice showed that antigenicity of E-binding domain fusion protein was almost as effective as E-hsp70 fusion protein and more effective than carrier protein hsp70 alone. In eliciting a humoral and cellular immune response, both fusion proteins were more powerful than the major antigenic segment of E protein alone, but less effective than the segment administered with Freund's adjuvant. 相似文献
2.
Immunization of cattle with synthetic peptides derived from the Boophilus microplus gut protein (Bm86) 总被引：6，自引：0，他引：6
Patarroyo JH Portela RW De Castro RO Pimentel JC Guzman F Patarroyo ME Vargas MI Prates AA Mendes MA 《Veterinary immunology and immunopathology》2002,88(3-4):163-172
Three synthetic peptides (SBm4912, SBm7462 and SBm19733), derived from the Bm86 glycoprotein from Boophilus microplus gut, were constructed and used to immunize cattle from a tick-free area. The immunized animals received three subcutaneous doses of the peptides, with saponin as adjuvant, at 30-day intervals. The immune response was evaluated by IgG elicited against the peptides by the detection of anti-Bm86 specific antibodies in situ and by Western blotting analysis. After tick challenge, reduction in the number, weight and oviposition capacity of engorged females was observed in the tick population that had fed on immunized animals. The results pointed a high efficacy (81.05%) for the SBm7462 synthetic peptide in relation to the others (p<0.01), demonstrating the efficiency of the immune response elicited by synthetic peptides to control the cattle tick B. microplus. 相似文献
3.
用酸对鼠伤寒沙门氏菌进行处理,制成裸细菌,作为空肠弯曲菌共同抗原(CA)的载体.用这种裸细菌和CA的复合物免疫小鼠,以间接ELISA和琼脂糖凝胶扩散沉淀反应检测免疫反应的变化及血清效价.结果表明,用这种方法免疫的小鼠,其血清抗体的ELISA滴度与对照组(福氏完全佐剂+CA组,CA组)相差不显著,但能与可溶性CA形成沉淀线,并含有IgM和IgG两类抗体;而对照组则不与CA出现沉淀反应,且只含IgM一类抗体.这表明,酸处理的沙门氏菌作为异种细菌抗原的载体,在改变机体对某种抗原的免疫类型上具有一定的作用. 相似文献
4.
Yang Y Chen J Li H Wang Y Xie Z Wu M Zhang H Zhao Z Chen Q Fu M Wu K Chi C Wang H Gao R 《Comparative immunology, microbiology and infectious diseases》2007,30(1):19-32
Interleukin-2 (IL-2) is vital to elicit and amplify the cellular and humoral immune responses to foreign antigens, which is extensively utilized in the control of infectious disease and treatment of various cancers. Porcine and murine IL-2 genes were, respectively, subcloned into VR1020, designated as VPIL-2 and VMIL-2, and then encapsulated in chitosan nanoparticles (CNP) prepared by ionic linkage. The BALB/c mice were intramuscularly co-administrated with chitosan-IL-2 nanoparticles (CNP-IL2) and paratyphoid vaccine to test the adjuvant effect of CNP-IL2. On day 35, the immunized mice were orally challenged with virulent Salmonella. The content of IgG, IgA, IgM, IL-2, IL-4, IL-6 and specific antibody titer as well as the number of immunocompetent cells were systematically analyzed in the vaccinated mice. The results revealed that the levels of immunoglobulins, cytokines, the specific antibodies, together with the numbers of lymphocytes significantly increased in vaccinated mice inoculated with CNP-VPIL2 in contrast with those with naked IL-2 plasmids and blank plasmids. The CNP-VPIL2 immunized mice exhibited higher humoral and cellular immune responses, less severe clinical signs and lesions of disease caused by the bacteria than the other groups after challenge. These findings suggest that CNP-VPIL2 has a significant enhancement effect on immune responses of mice, which results in better immunoprotection against Salmonella infection, indicating that CNP-VPIL2 could be employed as an effective immunoadjuvant to elevate immunity of animals to conventional vaccines. 相似文献
5.
小鼠对猪囊虫抗原基因TS76的免疫应答 总被引：3，自引：0，他引：3
将猪囊虫抗原基因 TS76的真核表达型质粒 VTS76单独或与 p UC18联合肌肉免疫注射于 BAL B/ c小鼠 ,以MTT比色法检测小鼠脾淋巴细胞 Con A刺激的增殖反应及 IL - 2的诱生活性 ,EL ISA方法检测免疫小鼠 Ig G总量和特异性抗体水平 ,常规法检测外周血免疫细胞数量的动态变化。结果发现 ,VTS76免疫小鼠各项细胞和体液免疫应答反应指数均比空白对照组小鼠显著提高 ;联合免疫组小鼠的细胞免疫应答和体液免疫应答反应指数均比 VTS76小鼠提高。由此可见 ,以 VTS76免疫小鼠可诱导其特异性细胞和体液免疫反应 ,而 p UC18又明显提高了 VTS76免疫小鼠的免疫应答水平 ,具有显著的免疫增强作用 相似文献
6.
Cholera toxin (CT) is a well-known mucosal adjuvant in mammals, but it does not give conclusive results in chickens. Cells from the chicken immune system may be insensitive to CT activity. Our results showed that intravenously administered CT had strong immunomodulatory effects on chicken antigen-specific T- and B-cell immune responses. Seven and eight days post-inoculation (p.i.), chickens immunized with KLH and CT exhibited a faster and higher specific proliferative response in the spleen after in vitro restimulation than chickens immunized with KLH alone. At the same time, the specific antibody response in serum was significantly higher, with a strong IgG enhancement and a peak of IgA in chickens immunized with KLH and CT. The anti-KLH splenic antibody response in vitro involved a significant increase in specific IgG and IgA isotypes when CT was used as adjuvant. In conclusion, as in mammals, systemic CT demonstrated strong adjuvant properties in chickens enhancing T-cell priming in vivo and, thus, leading to increased specific antibody production, including IgA. 相似文献
7.
Ruiz LM Orduz S López ED Guzmán F Patarroyo ME Armengol G 《Veterinary parasitology》2007,144(1-2):138-145
Plasmid pBMC2 encoding antigen Bm86 from a Colombian strain of cattle tick Boophilus microplus, was used for DNA-mediated immunization of BALB/c mice, employing doses of 10 and 50microg, delivered by intradermic and intramuscular routes. Anti-Bm86 antibody levels were significantly higher compared to control mice treated with PBS. In the evaluation of immunoglobulin isotypes, significant levels of IgG2a and IgG2b were observed in mice immunized with 50microg of pBMC2. Measurement of interleukine (IL) levels (IL-4, IL-5, IL-12(p40)) and interferon-gamma (IFN-gamma) in the sera of mice immunized with pBMC2 indicated high levels of IL-4 and IL-5, although there were also significant levels of IFN-gamma. Mice immunized with pBMC2 showed antigen-specific stimulation of splenocytes according to the incorporation of bromodeoxyuridine and IFN-gamma secretion. In all trials, mice injected intramuscularly with 50microg of pBMC2 presented the highest immune response. Moreover, cattle immunized with this DNA vaccine showed antibody production significantly different to the negative control. In conclusion, these results suggest the potential of DNA immunization with pBMC2 to induce humoral and cellular immune responses against B. microplus. 相似文献
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9.
为了探讨人参茎叶总皂苷(ginseng stem-leaf saponins,GSLS)联合亚硒酸钠(Na2SeO3,简称Se)对伪狂犬病病毒(PRV)灭活疫苗免疫的增强作用,本试验给小鼠口服GSLS后,接种添加了Se的PRV灭活疫苗,并检测免疫后小鼠血清中PRV gB抗体及其亚类(IgG1和IgG2a)水平、淋巴细胞增殖水平、淋巴细胞分泌细胞因子(IFN-γ、IL-2、IL-4和IL-6)水平以及野毒(fPRV)攻毒后小鼠的存活率。结果显示,与仅仅免疫PRV灭活疫苗的小鼠相比,口服GSLS联合疫苗中添加Se(GSLS-Se)能够显著提高小鼠血清中PRV gB抗体及其亚类(IgG1和IgG2a)水平,淋巴细胞增殖水平和细胞因子(IFN-γ、IL-2和IL-6)水平。此外,GSLS-Se还提高了免疫小鼠对野毒感染的抵抗力。结果表明,GSLS-Se对PRV灭活疫苗具有免疫增强作用,值得在本动物上进一步研究。 相似文献
10.
There is a need to produce a vaccine against Rhodococcus equi pneumonia in foals in which immunity against infection is largely based on a type 1, cell-mediated, immune response. The VapA protein of the virulence plasmid of R. equi is highly immunogenic. To assess the potential of vapA-DNA to produce immunity, C57BL/6 and BALB/c mice were immunized with a DNA vaccine constructed from vapA incorporated into pcDNA3.1. The plasmid construct expressed VapA in a COS-7 cell line. Intramuscular immunization of mice resulted in enhanced clearance of R. equi from the liver of intravenously challenged mice compared to non-immunized controls. This effect was more marked when pORF-IL-12, a plasmid expressing murine IL12, was included with the vaccine. Antibody developed to VapA, with an IgG2a response being more marked in mice immunized with pcDNA-vapA than in non-immunized or in mice immunized with the mixed vapA and IL-12 plasmid constructs. In conclusion, this study has shown for the first time that DNA immunization with vapA enhances the immune responses of mice against R. equi infection, that the IgG subisotype response is consistent with a type 1-based immune response, and that this can be enhanced by injection of the IL-12 gene. 相似文献
11.
【目的】构建非洲猪瘟病毒CAS19-01/2019株(GenBank登录号:MN172368.1)结构蛋白P72的合成肽疫苗,通过免疫小鼠评估合成肽疫苗的免疫效力。【方法】利用ProtParam、SOPMA等软件分析P72蛋白的理化性质与结构信息,通过ABCpred、SVMtrip、IEDB预测P72蛋白的T细胞与B细胞抗原表位,筛选出显著的表位多肽区域,合成多肽辅以弗氏佐剂腹腔注射免疫小鼠,检测免疫组小鼠产生的特异性抗体、T淋巴细胞亚群、脾脏淋巴细胞增殖、细胞因子白介素4(IL-4)、IL-2、γ干扰素(IFN-γ)、免疫球蛋白(IgG),从体液免疫与细胞免疫角度评估合成肽的免疫效力。【结果】综合分析得出P72蛋白是稳定性亲水蛋白,二级结构中α-螺旋、β-转角、延伸链、无规则卷曲分别占19.35%、5.42%、25.08%和50.15%。筛选出了P72蛋白的8个优势抗原表位,626－634、520－528、298－306、203－211位氨基酸处为T细胞抗原表位,587－606、232－251、110－129、39－58位氨基酸处为B细胞抗原表位。整合优势表位合成2个多肽P72-1与P72-2,首次免疫小鼠14 d时可检测到P72-1与P72-2的特异性抗体,首免后28 d达到最高值,其最高抗体效价分别为1∶25 600与1∶12 800;免疫后小鼠T淋巴细胞亚群CD4＋/CD8＋显著上升(P＜0.05);脾脏淋巴细胞增殖试验结果显示,免疫组淋巴细胞数量均极显著升高(P＜0.01);细胞因子IL-4、IL-2、IFN-γ含量均极显著增加(P＜0.01)。【结论】本研究成功研制2种合成肽疫苗,在免疫效力上P72-2高于P72-1,二者都能产生高水平的特异性抗体,刺激脾脏淋巴细胞增殖,诱导产生细胞因子IL-4、IL-2、IFN-γ,本研究为非洲猪瘟合成肽疫苗研制奠定技术基础。 相似文献
12.
流行性乙型脑炎病毒(JEV)是一种严重危害人畜健康的虫媒病毒。表面囊膜蛋白(E蛋白)是该病毒的主要结构蛋白。本研究利用原核表达的乙型脑炎病毒SA14-14-2株结构蛋白E蛋白作为免疫原,免疫6周龄BALB/c小鼠,采用淋巴细胞杂交瘤技术进行融合,经有限稀释法获得1株特异性针对JEV E蛋白的杂交瘤细胞,命名为5D4,经测定5D4单抗亚类属于IgG2a,轻链为κ链。经Western blot证实所得杂交瘤细胞分泌的抗体可特异地与JEV E蛋白结合。结果表明,所制备的抗JEV E蛋白的单抗对病原检测具有很高的特异性,为JEV准确快速的病原诊断和JEV感染相关的基础性研究提供了物质基础。 相似文献
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Barbey C Cauchard S Cauchard J Laugier C Hartke A Petry S 《Research in veterinary science》2012,93(1):172-176
Rhodococcus equi remains a significant pathogen, causing severe pneumonia in foals. The development of vaccines and serologic diagnosis could be greatly facilitated by studying the humoral immune response to this equine pathogen. In this study, a crude extract of R. equi ATCC 33701-secreted proteins combined with the Montanide® ISA70 adjuvant was found to be highly immunogenic in mice with the highest titer of 99,000 on day 42 after the first subcutaneous immunization. This immune response was dependent on the quantity of proteins injected and the presence of adjuvant. By dot-blotting, eight recombinant secreted proteins were identified to react strongly with sera from immunized mice. Of these eight proteins, four were detected as immunogenic only when administered in conjunction with adjuvant. This screening strategy led to the identification of promising new candidates for vaccine development. 相似文献
15.
Estrada A Katselis GS Laarveld B Barl B 《Comparative immunology, microbiology and infectious diseases》2000,23(1):27-43
We have identified saponins in the root of Polygala senega L., a plant indigenous to the Canadian prairies, which display immunopotentiation activity to protein and viral antigens. By two-step extraction and hemolytic activity-guided fractionation by silica flush chromatography six saponin fractions were generated and their HPLC profiles determined. Two dominant fractions, designated as PS-1 and PS-2, were tested for adjuvant activity in mice immunized with ovalbumin, and hens immunized with rotavirus. The resulting adjuvant activity was compared with that of Quil A saponin. The P. senega saponins increased specific antibody levels to the antigens, in both mice and hens. In mice, there was a preferential increase of the IgG2a subclass, and upon in vitro secondary antigen stimulation, high IL-2 and IFN-gamma levels were observed in spleen cell cultures from P. senega saponins-immunized animals. The saponins were tested for their toxicity by lethality in mice and were found to be less toxic at the same dose than their counterpart Quil A. The results of this study indicated the potential of P. senega saponins as vaccine adjuvants to increase specific immune responses. 相似文献
16.
本研究旨在筛选最佳的复合佐剂和口服疫苗诱饵组方,并将筛选出的复合佐剂口服免疫试验小鼠,检测免疫小鼠血清IgG和肠道IgA抗体效价。采用牛肉粉、鸡肉粉、鱼肉粉、奶粉、血粉为主要原料,以细菌脂多糖、氢氧化锌、枸杞多糖、甘露低聚糖为疫苗免疫佐剂,通过正交试验方法将复合免疫佐剂和口服疫苗诱饵与狂犬病SRV9病毒混匀后口服免疫试验小鼠,并对免疫动物血清IgG抗体效价进行检测。优化的复合佐剂组方是细菌脂多糖100 μg、氢氧化锌0.8 mg、枸杞多糖50 mg、甘露低聚糖10 mg,免疫小鼠血清IgG抗体含量为3.24 mg/L,肠道IgA抗体含量为1.56 ng/mL;在疫苗诱饵的投喂试验中,狼、狐狸、犬、猫对鱼肉味诱饵的采食率最高,其次是血味和奶味诱饵;诱饵与狂犬病SRV9病毒混匀后口服免疫小鼠28 d后,血清IgG平均值为1.20 U/L。本试验筛选出口服疫苗复合免疫佐剂,经筛选制备的鱼肉味诱饵具备高采食性,适合野生食肉类动物口服疫苗的制备。可增加狂犬病SRV9病毒的免疫效应,为进一步研发重组减毒狂犬病口服疫苗提供了必要的技术支撑。 相似文献
17.
There has been an increasing interest in finding new formulations that qualify as vaccine adjuvants, which must be safe, stable, and have the capacity to stimulate a strong immune response. In this study, a basic formulation of a water-in-oil-in-water (W/O/W) adjuvant CV13 was developed, and ginseng stem-leaf saponins (GSLS) were added as an immune booster into oil phase. The physicochemical properties of the adjuvant were tested. Furthermore, the immune activity and the adjuvant effects, as indicated by the foot-and-mouth disease virus (FMDV) antigen were evaluated. The results showed that CV13 was similar in appearance to ISA 206 and could package FMDV antigen into a stable W/O/W emulsion. The FMD vaccine prepared with CV13 alone or CV13 containing GSLS achieved pharmaceutical characteristics comparable to a vaccine prepared with ISA 206, moreover the structural stability of the CV 13 vaccine was found to be better. Mice that were immunized with the FMD vaccine prepared with CV13 containing GSLS presented a significantly higher LPBE antibody titer and splenocyte proliferation rate than those immunized with a vaccine prepared with CV13 alone (p < 0.05). In addition, there was no significant difference between the groups that were immunized with FMD vaccine prepared with CV13 containing GSLS and ISA206 in terms of cellular and humoral immune response. In this paper, CV13 containing GSLS shows excellent immunologic adjuvant effect in mice model, and this new adjuvant may provide a potential choice for FMD vaccine production in the future. 相似文献
18.
J L Abán V Ramajo J L Arellano A Oleaga G V Hillyer A Muro 《Veterinary parasitology》1999,83(2):107-121
Three strains of mice (NMRI, C57/BL, BALB/c) were each immunized with a 12 kDa purified, native Fasciola hepatica fatty acid binding protein (Fh12) and challenged percutaneously with Schistosoma bovis cercariae. C57/BL mice immunized with Fh12 had significant reductions in S. bovis worm burden recoveries (96 and 87% reductions over controls in two separate experiments). When using NMRI or BALB/c mice, Fh12 alone or in Freund's adjuvant failed to induce significant protection against S. bovis. In C57/BL mice vaccinated against Fh 12, antibodies to the IgG2a isotype, but not to the IgG1 isotype, increased by 2 weeks after the second immunization and remained high through 8 weeks of S. bovis infection. Antibodies to S. bovis increased after 4 weeks of infection. Regarding cytokine production by spleen mononuclear cells, C57/BL mice vaccinated with Fh12 in adjuvant, and having the highest protective response against challenge infection with S. bovis, had an increase of IFN-gamma production with Concanavalin A but no increase of IL-4 in similarly stimulated cells. These results suggest that the protection obtained in this group of mice is mediated by a Th1 immune response. 相似文献
19.
Chen D Colditz IG Glenn GM Tsonis CG 《Veterinary immunology and immunopathology》2000,77(3-4):191-199
Cholera (and related) toxins (CT) when applied topically on unbroken skin induce systemic immune responses in mice, a procedure called transcutaneous immunization (TCI). The current study examined the capacity for TCI to induce systemic immune responses in sheep. Three groups (n=5 per group) were immunized at day 0 (priming) and day 28 (boosting) with 250 microg of CT in water by TCI, with 25 microg of CT in alum by intramuscular injection, or not immunized. Serum samples were taken at days 0, 28, 42, 56 and 70 after immunization for measurement of CT-specific IgG as well as CT-specific IgG1, IgG2, IgA and IgM antibodies by ELISA. After immunization, IgG, IgG1 and IgG2 antibody in immunized groups were significantly higher than in the control group, and boosting further increased these titres. IgG, IgG1 and IgG2 in the injection group were significantly higher than in the TCI group. There was a preponderance of IgG1 antibody, relative to IgG2, in both immunized groups. CT-specific IgA and IgM were detected in both immunized groups. Lymphocyte proliferation to CT was measured at day 90. A CT-specific lymphocyte proliferative response (stimulation index>2) was detected in all sheep from the injection group, in two sheep from the TCI group and in none of the controls. Results demonstrated that TCI induces primary and secondary antibody responses and specific proliferative responses to CT in sheep. 相似文献