共查询到19条相似文献，搜索用时 250 毫秒
1.
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
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将40只BALB／c小鼠随机分为2组，A组以VR1020免疫作为对照，B组以TS2l抗原基因的真核表达型质粒VTS2l免疫。用ELISA检测免疫小鼠IgG总量和特异性抗体水平，MTT比色法检测小鼠脾淋巴细胞伴刀豆蛋白A(ConA)刺激的增殖反应及IL—2的谤生活性，常规法检测外周血免疫细胞数量的动态变化。结果显示，VTS2l免疫小鼠血清的IgG含量和特异性抗体效价显著高于对照组小鼠；免疫小鼠脾淋巴细胞ConA刺激增殖反应和IL—2诱生活性均比对照组小鼠显著增强；免疫小鼠的淋巴细胞、巨噬细胞等免疫细胞的数量也显著超过对照组。免疫小鼠的细胞和体液免疫反应显著增强，表明VTS2l具有很强的免疫激活作用，有进一步研制开发成为猪囊虫病DNA疫苗的潜力。 相似文献
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《中国预防兽医学报》2019,(1)
为提高滴鼻免疫条件下猪支原体肺炎活疫苗的免疫效果,本研究设计引物扩增猪肺炎支原体P97R1基因,融合至霍乱毒素B亚单位(CTB)基因下游,构建了重组质粒p ET-CTB-P97R1。重组菌经IPTG诱导表达、分离纯化得到r CTB-P97R1重组蛋白。利用体外神经节苷脂(GM1)结合试验检测r CTB-P97R1的佐剂活性。随后将r CTB-P97R1与活疫苗混合后滴鼻免疫小鼠,免疫后采血和收集小鼠支气管肺泡灌洗液(BALF),分别检测其中的特异性Ig G抗体和s Ig A抗体的水平评价免疫效果。结果显示,r CTB-P97R1可以特异性结合GM1,具有生物学活性。小鼠免疫后的血清Ig G抗体显著高于活疫苗对照组(p0.01),同时产生了高水平的P97R1特异性Ig G抗体(p0.01);小鼠BALF中的s Ig A抗体也显著高于活疫苗对照组(p0.05)。结果表明,r CTB-P97R1能够显著增强经滴鼻免疫的猪支原体肺炎活疫苗的免疫效力,同时可提升P97R1的免疫应答,有望作为经黏膜免疫的猪支原体肺炎活疫苗的新型佐剂。 相似文献
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为开发高效安全经济的免疫调节剂,本研究构建了共表达猪IL-4/6和猪抗菌肽融合基因VRP的重组毕赤酵母SG46P。通过猪淋巴细胞增殖试验和抑菌试验检测其发酵上清液的生物学活性和抑菌活性,并用该重组毕赤酵母对30只21日龄雌性ICR小鼠进行灌胃接种。接种后,每周采集小鼠尾部静脉血进行免疫功能分析试验;并在第28 d用大肠杆菌和金黄色葡萄球菌进行小鼠腹腔攻毒试验。结果显示:实验组SG46P的发酵上清液较对照组能显著刺激猪淋巴细胞的增殖(p0.05),且该发酵上清液具有明显的抑菌作用(p0.05)。接种重组毕赤酵母SG46P的小鼠的体质量有所增加(p0.05)且其外周血中白细胞数量、血红蛋白含量均显著高于对照组(p0.05);其血清中Ig G、Ig G1和Ig G2a抗体水平均较对照组显著增加(p0.05),其外周血中CD4+T和CD8+T淋巴细胞数量均显著高于对照组(p0.05);TLR4、TLR9、IL-2、IFN-γ、IL-4、IL-6、CD62L、IL-7、IL-23、CAMP和Crp4基因的转录水平均显著高于对照组(p0.05)。攻毒后,接种SG46P的小鼠的生存率显著高于对照组(p0.05)。结果表明:重组毕赤酵母SG46P不仅能够显著提高小鼠免疫基因的转录水平,有效提高动物的先天和获得性免疫水平,还可明显增强小鼠抗感染能力,这为研制高效安全经济的分子免疫调节剂和防治动物传染病开拓了新途径。 相似文献
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CpG ODN对传染性腔上囊病病毒活疫苗免疫效果的影响 总被引：2，自引：0，他引：2
将120只来航系SPF鸡随机分成对照组、注射CpG ODN组、IBDV活疫苗点眼免疫组及IBDV活疫苗 CpG ODN免疫组，每组30只，于7日龄时对试验鸡进行初次免疫，21日龄时进行加强免疫，28日龄时用IBDV野毒株攻毒。检测不同时间的IBDV特异性抗体效价、血液和脾淋巴细胞的Con A刺激指数(SI)。结果表明，CpG ODN处理后脾细胞的增殖转化能力强于血液淋巴细胞的增殖转化能力；CpG ODN的使用使血清中IBDV特异性抗体产生时间提前7d，抗体效价升高；与对照组相比，仅用CpG ODN处理就可使1／3的鸡攻毒后迅速产生高效价(约1：3000)的特异性抗体，降低了发病率和死亡率，证明CpG ODN能增强IBDV活疫苗的免疫效果。 相似文献
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SARS灭活疫苗的实验免疫初步研究 总被引：1，自引：0，他引：1
观察 SARS灭活疫苗在实验动物体内的免疫效果 ,初步探讨 SARS灭活疫苗的免疫机理。方法 :利用 Vero E6细胞培养 SARS病毒 ,加入甲醛将其灭活 ,以此为抗原 ,筛选合适佐剂 ,制定合理的免疫程序 ,分别免疫 BAL B/ c小鼠、C57BL/ 6J小鼠及 SD大鼠 ,免疫后每两周采外周血一次 ,用流式细胞仪测外周血 CD4 、CD8 ,计算淋巴细胞总数。同时用 EL ISA法及中和抗体法测定抗 SARS冠状病毒 Ig G抗体的水平。结果显示 :与对照组比较 ,SARS灭活疫苗进行免疫后的实验动物外周血淋巴细胞的百分比计数、CD4 / CD8 T淋巴细胞之间的比值随着时间的变化均有不同程度的增长 ;Ig G中和抗体水平达到 1∶ 2 560 ,EL ISA抗体水平达到 1∶40 960。表明 SARS灭活疫苗可以有效刺激实验动物体内 T淋巴细胞、B淋巴细胞的活化过程 ,能成功诱导细胞免疫和体液免疫 ;SARS灭活疫苗同时还具有较强的免疫原性 ,可刺激实验动物产生具有免疫保护作用的特异性 Ig G抗体 相似文献
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表达猪圆环病毒2型cap蛋白的重组罗伊氏乳酸杆菌在小鼠诱导的免疫应答分析 总被引：1，自引：1，他引：0
旨在构建表达猪圆环病毒2型（PCV2）cap蛋白的重组罗伊氏乳酸杆菌（Lactobacillus reuteri,L.reuteri）,并评价其在小鼠体内诱导的免疫应答效果。利用PCR扩增实验室分离保存的PCV2b型毒株的cap蛋白基因,以猪源L.reuteri为宿主菌,构建表达cap蛋白的重组菌株pPG-T7 g10-PPT-cap/L.reuteri,通过口服免疫BALB/c小鼠。采用间接ELISA方法测定免疫后小鼠血清中抗原特异性IgG抗体水平,粪便、鼻腔洗液、生殖道洗液、肠黏液中抗原特异性sIgA抗体水平,小鼠血清中各细胞因子水平;MTT法检测小鼠脾淋巴细胞增殖水平;流式细胞技术检测小鼠脾淋巴细胞中CD4+T细胞、CD8+T细胞的水平;荧光定量PCR检测免疫后攻毒的小鼠体内器官的病毒载量。结果显示,口服免疫重组乳酸菌组小鼠血清IgG抗体水平显著高于对照组（P<0.01）;小鼠粪便、鼻腔洗液、生殖道洗液、肠黏液中sIgA抗体水平显著高于对照组（P<0.01）;小鼠血清中细胞因子水平和对照组相比,IFN-γ、IL-2、IL-4、IL-12水平升高,IL-10水平降低,IFN-α无显著变化;体外孵育PCV2和小鼠脾淋巴细胞结果表明,重组乳酸菌组小鼠脾淋巴细胞增殖刺激指数显著高于对照组（P<0.01）;流式细胞技术检测结果显示,口服免疫重组乳酸菌组小鼠脾细胞中CD4+T细胞、CD8+T细胞含量高于对照组;荧光定量PCR结果显示,相比于对照组,口服免疫重组乳酸菌组小鼠体内的病毒载量明显低于对照组。综上所述,本研究成功构建了表达PCV2 cap蛋白的重组罗伊氏乳酸杆菌,经口服途径免疫动物,构建的重组乳酸杆菌能够刺激小鼠产生体液免疫和细胞免疫应答,且具有一定的免疫保护效果。 相似文献
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HBV转基因小鼠T细胞免疫状态 总被引：2，自引：0，他引：2
以正常非转基因 ICR小鼠为对照 ,用 3H-Td R掺入法和 ELISA分别测定了 HBV转基因 ICR小鼠脾淋巴细胞针对 HBe Ag的特异性增殖功能及其培养液上清中 Th1、Th2相关细胞因子 m IFN-γ、m IL-2、m IL-6、m IL-1 0含量。结果 ,HBV转基因小鼠脾淋巴细胞对 HBe Ag刺激引起的增殖反应、HBe Ag与 Con A共同刺激引起的增殖反应及其培养液上清中 4种细胞因子含量均明显低于对照组。以上结果表明 ,HBV转基因小鼠 Th1、Th2淋巴细胞所介导的特异性免疫功能均受到抑制 相似文献
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细胞凋亡抑制基因免疫对小鼠生长及生长激素的影响 总被引：1，自引：0，他引：1
本实验用细胞凋亡抑制基因 (bcl- 2 )JLV重组质粒 (JLV -bcl)免疫小鼠 ,测定免疫后小鼠的生长及血浆GH浓度。结果表明 ,经bcl- 2免疫后实验组小鼠体重均低于对照组 ,在 7～ 8周龄时实验A组小鼠体重与对照组存在显著差异 (P <0 .0 5)。用RIA双抗法检测血浆中生长激素 (GH)浓度 ,发现在首免后 3周实验A组小鼠的GH浓度显著低于对照组 (P <0 .0 5)。这些结果均提示bcl- 2基因与小鼠生长有关 ,如果用基因治疗的方法增加bcl- 2基因表达量可以增加小鼠的生长速度 相似文献
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为了研究猪IL-2基因的真核质粒和融合基因形式对pcDNA—E39．VP2真核共表达质粒的免疫增强作用,分别将IL-2基因插入pcDNA3．1（＋）和pcDNA—E39．VP2（已构建）中构建重组质粒pcDNA-IL2和pcDNA-IL2-E39．VP2。将真核质粒pcDNA-IL2-E39．VP2免疫小鼠,另外,将pcDNA—IL2分别先于和后于pcDNA-E39．VP23d免疫小鼠,同时设pcDNA-E39．VP2对照组。对免疫小鼠进行PPV（JEV）抗体水平、脾脏淋巴细胞增殖功能和外周血T淋巴细胞亚群变化的检测。结果显示,1周后,pcDNA-IL2-E39．VP2免疫组和pcDNA—IL2后于pcDNA—E39．VP注射的免疫组JEV／PPV抗体水平和脾脏淋巴细胞增殖活性显著（P〈0．05）或极显著（P〈0．01）高于对照组,pcDNA-IL2先于pcDNA—E39．VP注射的免疫组仅在第5周显著（P〈0．05）高于对照组;各组（除对照组外）CD8^＋值差异不显著（P〉0．05）,CD4^＋、CD4／cD8值都高于对照组,其中pcDNA-IL2-E39．VP2免疫组与对照组差异极显著（P〈0．01）。结果表明,IL-2能增强pcDNA-E39．VP诱导小鼠的免疫能力,且以融合基因形式构建的真核质粒免疫增强效果最明显。 相似文献
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试验旨在构建表达猪附红细胞体ENO基因的质粒DNA,并测定其免疫效果。将猪附红细胞体ENO基因克隆到PVAX1真核表达载体上,然后转染到Vero细胞中进行表达并测定其免疫效果。将18只BALB/c小鼠（雌雄各半）随机分为3组（PVAX1-ENO质粒DNA免疫组、PVAX1空载体对照组及PBS对照组）,每组6只,每组小鼠对应免疫接种PVAX1-ENO质粒DNA、PVAX1空质粒和PBS。分离血清并通过ELISA方法测定血清中ENO蛋白抗体效价、IgG1和IgG2a抗体水平及IFN-γ和IL-4细胞因子水平。三免2周后每组选取3只小鼠检测CD4+和CD8+含量。结果显示,本试验成功构建了PVAX1-ENO质粒DNA,经PCR和酶切鉴定正确,并能在Vero细胞中成功表达。PVAX1-ENO质粒DNA免疫组ENO蛋白抗体水平、IgG1和IgG2a抗体水平、IFN-γ和IL-4细胞因子水平及CD4+和CD8+含量均显著高于PVAX1空载体对照组及PBS对照组（P<0.05）。结果表明,PVAX1-ENO质粒DNA可显著提高BALB/c小鼠的体液免疫水平,并在一定程度上刺激BALB/c小鼠细胞免疫。 相似文献
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为观察姬松茸水溶性多糖对镉中毒小白鼠血液系统的治疗作用,将30只SPF级雄性健康小白鼠随机分为对照组、试验1组、试验2组共三组,两个试验组腹腔注射2.5 mg/（kg·d）氯化镉溶液染毒,对照组腹腔注射与其等容量的生理盐水,每周连续4 d,造模5周,之后对试验2组用300 mg/（kg·d）姬松茸多糖连续治疗15 d。结果表明,与试验1组相比,试验2组的血液指标RBC、HGB、MCV、MCH、MCHC均显著提高,而WBC、Mid、Grn则显著下降;与对照组相比,试验2组的血液指标HGB、MCV均显著降低,而Lym则显著升高。证实,姬松茸水溶性粗多糖对镉造成的血液系统的损伤具有一定程度的治疗作用。 相似文献
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Zhao ZZ Zhang HB Chen Q Su D Xie Z Wang YY Yang Y Wang ZZ Li JL Wu KY Wang HN Meng MJ Gao R 《Comparative immunology, microbiology and infectious diseases》2009,32(3):191-205
A novel oligodeoxynuleotides containing 11 CpG motifs was synthesized and inserted into the VR1020 plasmid containing pig interleukin-6 (IL-6) gene (VPIL6) to construct recombinant plasmid, VPIL6C. The chitosan nanoparticles (CNP) were prepared by ionic cross linkage to entrap the VPIL6C (VPIL6C-CNP), VPIL6 (VPIL6-CNP) and CpG (CpG-CNP). 42-Day old female mice were divided into four groups and intramuscularly injected respectively with 6 pmol VPIL6C-CNP, VPIL6-CNP, CpG-CNP and VR1020-CNP along with the bivalent vaccines against the Pasteurellosis and hog cholera. The blood was weekly collected from mice after vaccination to detect the changes of immunoglobulins, specific antibodies, IL-2, IL-4, IL-6 and immune cells. 28 days after vaccination, the mice were orally challenged with virulent Pasteurella multocida. The results showed that in comparison with those of the control VR1020 group, the content of immunoglobulins, specific antibodies and interleukins significantly increased in the sera from the treated two groups (P<0.05). Meanwhile, the number of lymphocytes and monocytes also remarkably elevated in the treated groups (P<0.05). The immune responses of VPIL6C mice were notably stronger than those of VPIL6 and CpG group. The challenge results proved that the overall immunity was further promoted in the treated mice which resisted the challenge infection; while the control mice manifested evident symptoms and lesions, and died of infection. These suggested that VPIL6C-CNP could better promote the immunity and resistance of mice against Pasteurellosis than VPIL6-CNP and CpG-CNP, and facilitate the development of effective adjuvant to enhance the immunity of animal against infection. 相似文献
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DNA immunization against very virulent infectious bursal disease virus with VP2-4-3 gene and chicken IL-6 gene 总被引：2，自引：0，他引：2
Sun JH Yan YX Jiang J Lu P 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2005,52(1):1-7
The present study was to investigate the feasibility and efficiency of the DNA vaccine to protect chickens against very virulent infectious bursal disease virus (vvIBDV) infection. A plasmid DNA carrying VP2-4-3 genes of vvIBDV SH95 and a plasmid DNA carrying chicken interleukin-6 (ChIL-6) genes were constructed and designated as pALTER-MAX-VP2-4-3 and pALTER-MAX-ChIL-6 respectively. Several DNA vaccination experiments were performed: 1-week-old chickens were intramuscularly injected with only plasmid pcDNA3-VP2, pALTER-MAX-VP2-4-3 or mixture with pALTER-MAX-ChIL-6. The chickens at 4 weeks old were orally inoculated with vvIBDV SH95. The results showed that immunization with the mixture of pALTER-MAX-VP2-4-3 and pALTER-MAX-ChIL-6 three times conferred protection for 90% of chickens. Enzyme-linked immunosorbent assay (ELISA) antibody titres in chickens immunized together with pALTER-MAX-ChIL-6 were higher than those immunized simply with plasmid pcDNA3-VP2 or pALTER-MAX-VP2-4-3. IBDV was not detected in the bursa of the protected chickens at 8 days after challenge by RT-PCR. The results indicate that protection against vvIBDV can be achieved by using the VP2-4-3 gene of vvIBDV as a DNA vaccine. Furthermore, the simultaneous injection of ChIL-6 plasmid significantly increased the protection after challenge with the very virulent strain. 相似文献
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Regulating effects of porcine interleukin-6 gene and CpG motifs on immune responses to porcine trivalent vaccines in mice 总被引：5，自引：0，他引：5
Wu M Gao R Meng M Li J Tan M Shen Y Wang L Yin X Wu X Xie H Liu S 《Research in veterinary science》2004,77(1):49-57
In order to develop novel immunoadjuvants to boost immune response of conventional vaccines, experiments were conducted to investigate the regulating effects of porcine interleukin-6 gene and CpG motifs as the molecular adjuvants on immune responses of mice that were co-inoculated with trivalent vaccines against Swine fever, the Pasteurellosis and Erysipelas suis. Synthetic oligodeoxynuleotides containing CpG motifs were ligated into pUC18, forming recombinant pUC18-CpG plasmid. Eukaryotic plasmid expressing porcine interleukin-6 (VPIL-6) were also constructed as molecular adjuvants in an attempt to enhance levels of immune responses of mice co-administered with the trivalent vaccines in this paper. The cellular and humoral immune responses of mice were systematically analysed, and the experimental results were observed that the number of white blood cells, monocytes, granuloytes and lymphocytes significantly increased, respectively, in the mice immunized with VPIL-6, compared with those of the control; the IgG content and titre of specific antibodies to the trivalent vaccine mounted remarkably in the sera from the VPIL-6 vaccinated mice; the proliferation of lymphocytes and induced IL-2 activities were significantly increased in the vaccinated groups. The above-mentioned immune responses of mice co-inoculated with pUC18-CpG plasmid were significantly stronger than those of co-inoculated with pUC18 plasmid, suggesting that the immunostimulatory effect of oligodeoxynuleotides CpG is closely connected with the number of CpG motifs. These results suggest that the porcine IL-6 gene and CpG motifs could be employed as effective immunoadjuvants to elevate immunity to conventional vaccines. 相似文献
18.
为了研究猪IL-2基因的真核质粒和融合基因形式对pcDNA-E39.VP2真核共表达质粒的免疫增强作用,分别将IL-2基因插入pcDNA3.1(+)和pcDNA-E39.VP2(已构建)中构建重组质粒pcDNA-IL2和pcDNA-IL2-E39.VP2。将真核质粒pcDNA-IL2-E39.VP2免疫小鼠,另外,将pcDNA-IL2分别先于和后于pcDNA-E39.VP2 3d免疫小鼠,同时设pcDNA-E39.VP2对照组。对免疫小鼠进行PPV(JEV)抗体水平、脾脏淋巴细胞增殖功能和外周血T淋巴细胞亚群变化的检测。结果显示,1周后,pcDNA-IL2-E39.VP2免疫组和pcDNA-IL2后于pcDNA-E39.VP注射的免疫组JEV/PPV抗体水平和脾脏淋巴细胞增殖活性显著(P<0.05)或极显著(P<0.01)高于对照组,pcDNA-IL2先于pcDNA-E39.VP注射的免疫组仅在第5周显著(P<0.05)高于对照组;各组(除对照组外)CD8+值差异不显著(P>0.05),CD4+、CD4/CD8值都高于对照组,其中pcDNA-IL2-E39.VP2免疫组与对照组差异极显著(P<0.01)。结果... 相似文献
19.
The present study was to investigate the feasibility and efficiency of the DNA vaccine to protect chickens against very virulent infectious bursal disease virus (vvIBDV) infection. A plasmid DNA carrying VP2‐4‐3 genes of vvIBDV SH95 and a plasmid DNA carrying chicken interleukin‐6 (ChIL‐6) genes were constructed and designated as pALTER‐MAX‐VP2‐4‐3 and pALTER‐MAX‐ChIL‐6 respectively. Several DNA vaccination experiments were performed: 1‐week‐old chickens were intramuscularly injected with only plasmid pcDNA3‐VP2, pALTER‐MAX‐VP2‐4‐3 or mixture with pALTER‐MAX‐ChIL‐6. The chickens at 4 weeks old were orally inoculated with vvIBDV SH95. The results showed that immunization with the mixture of pALTER‐MAX‐VP2‐4‐3 and pALTER‐MAX‐ChIL‐6 three times conferred protection for 90% of chickens. Enzyme‐linked immunosorbent assay (ELISA) antibody titres in chickens immunized together with pALTER‐MAX‐ChIL‐6 were higher than those immunized simply with plasmid pcDNA3‐VP2 or pALTER‐MAX‐VP2‐4‐3. IBDV was not detected in the bursa of the protected chickens at 8 days after challenge by RT‐PCR. The results indicate that protection against vvIBDV can be achieved by using the VP2‐4‐3 gene of vvIBDV as a DNA vaccine. Furthermore, the simultaneous injection of ChIL‐6 plasmid significantly increased the protection after challenge with the very virulent strain. 相似文献