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1.
CpG ODN对猪繁殖障碍与呼吸道综合征疫苗免疫影响的研究   总被引:4,自引:1,他引:4  
以猪繁殖障碍与呼吸道综合征(蓝耳病.PRRS)病毒为模式病毒.以CpG ODN联合猪繁殖障碍与呼吸道综合征灭活疫苗免疫仔猪.通过ELISA法检测猪血清中抗猪繁殖障碍与呼吸道综合征病毒抗体水平,并用MTT法检测猪淋巴细胞白介素-2(IL-2)诱生活性以及淋巴细胞增殖指数(SI).试验结果表明:CpG ODN能显著提高仔猪的特异性抗体滴度、淋巴细胞IL-2诱生活性以及淋巴细胞增殖反直.证明CpG ODN能显著增强猪对常规灭活病毒疫苗的免疫应答能力。  相似文献   

2.
将CpGODN、non CpG ODN分别接种初生仔猪,同时分离仔猪的外周血单核细胞,以CpGODN、non CpG ODN对外周血单核细胞体外培养,分别检测仔猪Th1型细胞因子分泌情况。结果表明,与non CpG ODN和对照组相比,CpG ODN能在体内显著提高仔猪的淋巴细胞白介素-2诱生活性、淋巴细胞增殖、血清中IFN-γ和IL-12水平(P〈0.05),亦能在体外刺激仔猪的外周血单核细胞分泌IFN-γ和IL-12(P〈0.01)。这显示CpG—ODN能显著增强动物的免疫应答能力。  相似文献   

3.
如何提高和改善疫苗的免疫效果是目前疫苗研究的热点,近年来许多研究发现与DNA疫苗联合应用的细胞因子可通过不同的环节调节和增强DNA疫苗免疫效应,白细胞介素作为免疫佐剂能协同DNA疫苗能取得较好的免疫效果。细胞因子是动物体内特异高效调控免疫细胞和协调免疫应答重要的信号分子,它主要包括淋巴毒素(LT)、肿瘤坏死因子(TNF)、干扰素(INF)、白细胞介素(IL,如IL-2、IL-4、IL-5、IL-6、IL-7、IL-8、IL-12)和集落刺激因子(GM-CSF)等。其中IL-2和IL-4对病毒保护性抗原基因真核表达影响的研究较多。主要综述了IL-2和IL-4作为免疫佐剂对病毒保护性抗原基因真核表达的影响,以期为更深入地进行IL-2和IL-4对DNA疫苗免疫增强作用的深入研究提供参考。  相似文献   

4.
本试验通过分子克隆技术分别构建了单独表达猪繁殖与呼吸综合征病毒(PRRSV)GP5基因以及PRRSVGP5基因和猪IL-18基因共表达的重组核酸疫苗质粒(pEGFP—GP5和pEGFP—ILl8-GP5),并进行仔猪免疫原性研究,对构建的PRRSV核酸疫苗所诱导的体液免疫和细胞免疫水平进行检测,进一步研究了PRRSV核酸疫苗免疫效果以及猪IL-18基因对PRRSV核酸疫苗免疫调节作用的影响。同时,调查了商业上应用不同类型的PRRSV疫苗诱导的免疫效果,并与核酸疫苗免疫效果进行比较。结果表明,IL-18作为免疫佐剂在疫苗免疫猪后诱导的病毒特异性细胞免疫反应方面具有很好的调节作用,共表达IL18-GP5蛋白能够明显的改善DNA疫苗的免疫效力,增强抗PRRSV的免疫保护。因此,DNA疫苗做为一种新一代疫苗可用于对抗高致病性PRRSV感染。  相似文献   

5.
CpG DNA重组质粒对猪O型口蹄疫病毒抗原的免疫佐剂效应   总被引:1,自引:0,他引:1  
作者应用含CpG基序序列的重组质粒(即CpG DNA)作为免疫佐剂,评价其对猪O型口蹄疫病毒抗原疫苗的免疫增强效果.结果表明:CpG DNA重组质粒与猪口蹄疫灭活病毒抗原疫苗配伍免疫小鼠,CpG DNA重组质粒对小鼠具有较强的免疫佐剂效应,能促进口蹄疫病毒抗原诱导产生较高水平的特异性抗体,其抗体滴度是空载体疫苗对照的2倍.CpG DNA重组质粒与商品化猪口蹄疫灭活疫苗配伍免疫试验猪,其增强抗原诱导的特异性抗体滴度最高可达标准疫苗的4倍以上,也显著高于空载体疫苗对照;与病毒纯化抗原配伍免疫动物,攻毒后其免疫保护效力的PD50高达13.00,远高于标准疫苗对照(PD50> 4.69).在CpG DNA重组质粒剂量选择试验中,含低剂量的CpG DNA疫苗(50、200μg·头份-1)都比高剂量组(500 μg·头份-1)诱导的抗体滴度高,其中50和200μg·头份-1的CpG DNA剂量,在接种后14~32 d诱导的抗体滴度高达1:1 500,是标准疫苗的4~5倍,而500μg·头份-1剂量诱导的抗体仅是标准疫苗的2倍,说明CpG DNA重组质粒在低剂量时即可发挥强烈的佐剂效应.研究表明CpG DNA对猪O型口蹄疫病毒抗原疫苗有较强的免疫佐剂效应,且使用剂量低,应用前景广阔.  相似文献   

6.
为了探讨CpG寡核苷酸(CpG oligonucleotide,CpG ODN)对鸡新城疫疫苗免疫效力的影响,将CpG2007与鸡淋巴细胞共孵育,测定淋巴细胞增殖率,结果发现CpG2007对鸡淋巴细胞具有显著的刺激活性。将CpG2007与不同浓度的新城疫抗原混合,制备灭活疫苗,免疫健康雏鸡。分别于免疫后不同时间采血,测定抗体效价和细胞因子表达量,并进行攻毒保护试验。结果发现,添加CpG ODN佐剂的试验组均比对应相同抗原剂量的免疫对照组的抗体水平高,产生抗体速度快;抗原剂量降低10倍的佐剂试验组与高抗原剂量免疫对照组抗体水平和攻毒保护率均相当,表明CpG ODN能显著增强新城疫疫苗的免疫效力,能促进机体产生更强烈的免疫应答,是有效的疫苗佐剂候选物质。  相似文献   

7.
人工合成的含非甲基化CpG基序的寡聚脱氧核苷酸(CpG ODN)能够与树突状细胞、B细胞、单核细胞及巨噬细胞等免疫细胞内质网膜上的Toll样受体9(TLR-9)结合,通过产生Th1型免疫反应和分泌促炎因子来增强免疫应答的强度,在机体抵抗病原入侵和抗肿瘤等方面起到免疫增强剂的作用。近年来,CpG ODN作为一种新型免疫佐剂受到了人们越来越多的关注。论文主要介绍了CpG ODN的种类及其结构和功能特征、作用机制、对畜禽的免疫增强作用、安全性等方面的研究进展,为CpG ODN作为疫苗佐剂的进一步研究和在兽医临床上的应用提供参考。  相似文献   

8.
将提取的牛脾转移因子分别与猪瘟、猪繁殖与呼吸综合征疫苗混合后免疫30-32日龄仔猪,结果表明,提取的转移因子能显著增强猪瘟、猪繁殖与呼吸综合征疫苗的免疫效果,与对照组相比。抗体水平差异显著(P〈0.05)。  相似文献   

9.
白细胞介素-2(IL-2)是免疫学研究的重细胞因子之一,在动物疾病防治中发挥重作用,显示了良好的应用前景。就IL-2作为免疫佐剂增强疫苗免疫效果和提高动物抗感染性疾病能力等方面进行了综述,并对应用中存在的问题进行了探讨。  相似文献   

10.
为探究不同类型CpG基序对细粒棘球蚴(Echinococus granulosus)Eg95抗原的免疫增强作用,将pET-32a-3Eg95重组大肠杆菌,经IPTG诱导表达,亲和层析纯化重组蛋白pET-32a-3Eg95。将重组蛋白分别与常规佐剂Quli-A、pUC18-CpG和CpG ODN混合制备基因工程亚单位疫苗样品,免疫小鼠。通过间接ELISA方法检测抗体水平,通过实时荧光定量PCR方法测定体外刺激试验后细胞因子的相对表达量,检测不同佐剂在体液免疫和细胞免疫方面对Eg95抗原免疫效果的增强作用。结果显示,诱导表达重组蛋白的大小约为56 ku,与理论值相符,用羊细粒棘球蚴阳性血清检测有特异性条带;pUC18-CpG组和CpG ODN组小鼠免疫后14、28、42 d的抗体平均水平(D450 nm)分别为3.10、3.03、3.22和2.98、3.12、3.27,与Quli-A组抗体平均水平相比均差异显著(P<0.05);CpG ODN组血清中抗体平均水平略高于pUC18-CpG组,但差异不显著(P>0.05)。重组蛋白刺激后pUC18-CpG组与CpG ODN组IFN-β、IFN-γ和TNF-α的相对表达量分别为6.88、2.35、6.28和5.03、2.85、7.07,与Quli-A组相比差异均显著(P<0.05)。因此相对于常规佐剂Quli-A,pUC18-CpG和CpG ODN在体液免疫和细胞免疫方面对Eg95抗原都有显著的免疫增强作用,CpG ODN的免疫增强作用略优于pUC18-CpG,二者均可作为免疫佐剂,增强现有包虫病基因工程疫苗抗原的免疫效果。  相似文献   

11.
Oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) prevent development of T-helper type 2 (Th2) immune response and reverse established allergic responses in mouse models. However, little work on immune responses in piglets has been conducted in vivo. In this report, the ability of a porcine-specific CpG ODN to act as an immunostimulant and enhance immune responses of piglets to swine Pasteurella multocida living vaccine (SPML vaccine) was determined. The titre of IgG and IgG1/IgG2 isotype to SPML vaccine in serum, the proliferation of lymphocytes, SPML-specific interferon-gamma (IFN-gamma) and IL-6, TNF-alpha, IL-4 production of PBMCs in vitro and IFN-gamma, IL-6, TNF-alpha, IL-4, IL-10 in piglets serum were examined to identify the immune responses of the piglets. Immune responses of the piglets vaccinated with SPML and CpG ODN were significantly stronger than responses of piglets vaccinated with SPML alone. All these data summarized that immunostimulatory CpG ODN could modulate the immune response towards a Th1-like response when co-administered to piglets during SPML vaccination, which suggested that the therapeutic uses envisioned for these ODNs (as vaccine adjuvants and immunoprotective agents) may be applicable to husbandry animals.  相似文献   

12.
This study investigated the immunoadjuvant effects of three types of bacterial genomic DNA and CpG oligonucleotides (CpG ODN) on the avian influenza virus (AIV) subtype H5N1 inactivated oil emulsion vaccine under two immunization strategies. The genomic DNA extracted from Escherichia coli O2, Staphylococcus aureus, Streptococcus faecalis FQ68, and synthetic CpG ODN were used as adjuvants, and their effects on the AIV oil emulsion vaccine were examined in chickens. The results indicated that when administered separately from the vaccine, adjuvants induced lower haemagglutination inhibition (HI) titres and serum IgG titres but resulted in higher concentrations of IFN-γ and IL-10. In contrast, when combined with the oil emulsion vaccine prior to inoculation, CpG ODN induced higher HI, IgG titres and IFN-γ concentration but resulted in lower IL-10 concentration. These data suggest that, depending on the immunization approaches, adjuvants may exert distinct immune effects in chickens receiving AIV H5N1 oil emulsion vaccine: the prior incorporation of CpG ODN into the vaccine may augment both the humoral and Th1 type immune responses, while separate inoculation of adjuvants has not shown better adjuvanticity.  相似文献   

13.
胞嘧啶-鸟嘌呤寡脱氧核苷酸(cytosine phosphate guanidine oligodeoxynucleotide,CpG ODN)是指含有非甲基化的胞嘧啶和鸟嘌呤二核苷酸为核心序列的核苷酸序列,近年来,CpG ODN作为一种新型免疫佐剂的研究越来越多,可诱发机体产生多种免疫学效应,提高系统免疫和黏膜免疫水平,具有安全性高,耐受性强等特点。  相似文献   

14.
Immunostimulatory CpG oligodeoxynucleotides (ODN) have been tested as immunoadjuvants for various vaccines in mice and human. Findings from previous reports suggest that CpG ODN can be used to enhance magnitude and balance of an immune response while reducing undesirable side effects of commercial vaccine, when delivered by parenteral route. Recently, it has been showed that CpG ODN is a promising mucosal adjuvant in mice, but data on mucosal immune responses induced by CpG ODN in other animals, especially in chickens, are scarce. Herein, we evaluated intranasal (IN) delivery of CpG ODN with newcastle disease (ND) vaccine (NDV) to determine its potential as a mucosal adjuvant to a commercial vaccine. CpG ODN augmented systemic (IgG in serum, T cell proliferation) and mucosal (IgA in intestinal washings and feces) immune responses against antigen. CpG ODN stimulated effectively both systemic and mucosal immune responses when delivered intranasally. Results from this study indicate that stimulatory CpG ODN is a potential effective mucosal adjuvant for the NDV in SPF chickens and may be applicable to husbandry animals.  相似文献   

15.
Immunostimulatory CpG oligodeoxynucleotides (ODN) have been tested as immunoadjuvants for various vaccines in mice and human. Findings from previous reports suggest that CpG ODN can be used to enhance magnitude and balance of an immune response while reducing undesirable side effects of commercial vaccine, when delivered by parenteral route. Recently, it has been showed that CpG ODN is a promising mucosal adjuvant in mice, but data on mucosal immune responses induced by CpG ODN in other animals, especially in chickens, are scarce. Herein, we evaluated intranasal (IN) delivery of CpG ODN with newcastle disease (ND) vaccine (NDV) to determine its potential as a mucosal adjuvant to a commercial vaccine. CpG ODN augmented systemic (IgG in serum, T cell proliferation) and mucosal (IgA in intestinal washings and feces) immune responses against antigen. CpG ODN stimulated effectively both systemic and mucosal immune responses when delivered intranasally. Results from this study indicate that stimulatory CpG ODN is a potential effective mucosal adjuvant for the NDV in SPF chickens and may be applicable to husbandry animals.  相似文献   

16.
将猪瘟弱毒脾淋疫苗、猪O型口蹄疫灭活疫苗(Ⅱ)和高致病性猪蓝耳病灭活疫苗采用两两组合的方式同时分部位免疫35日龄左右的健康仔猪,观察并记录其免疫反应。在疫苗免疫后0、4、14、21d和28d无菌取猪血清,用ELISA检测免疫后三种疫苗抗体水平的动态变化。试验结果表明:猪瘟和猪口蹄疫疫苗同时免疫,效果较好;猪口蹄疫和高致病性猪蓝耳病疫苗同时免疫,效果较差;猪瘟疫苗和高致病性猪蓝耳病疫苗同时免疫,效果不明显,还需进一步验证。  相似文献   

17.
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.
Culture filtrate protein (CFP) vaccines have been shown to be effective in small animal models for protecting against tuberculosis while immunisation with these types of vaccines in cattle has been less successful. A study was conducted in cattle to evaluate the ability of selected adjuvants and immunomodulators to stimulate protective immune responses to tuberculosis in animals vaccinated with Mycobacterium bovis CFP. Seven groups of cattle (n=5) were vaccinated with M. bovis CFP formulated with either Emulsigen or Polygen adjuvant alone or in combination with a specific oligodeoxynucleotides (ODN), polyinosinic acid: polycytidylic acid (poly I:C) or poly I:C and recombinant granulocyte-macrophage colony stimulating factor. Two additional groups were vaccinated subcutaneously with BCG or non-vaccinated. In contrast to the strong interferon-gamma (IFN-gamma) responses induced by BCG, the CFP vaccines induced strong antibody responses but weak IFN-gamma responses. The addition of CpG ODN to CFP significantly enhanced cell-mediated responses and elevated antibody responses to mycobacterial antigens. Of the CFP vaccinated groups, the strongest IFN-gamma responses to CFP vaccines were measured in animals vaccinated with CFP/Emulsigen+CpG or CFP/Polygen+CpG. The animals in these two groups, together with those in the BCG and non-vaccinated groups were challenged intratracheally with virulent M. bovis at 13 weeks after the first vaccination and protection was assessed, by examination for presence of tuberculous lesions in the lungs and lymph nodes, 13 weeks later at postmortem. While BCG gave the best overall protection against tuberculosis, significant protection was also seen in animals vaccinated with CFP/Emulsigen+CpG. These results establish an important role for CpG ODN in stimulating protective Th1 responses to tuberculosis in cattle and indicate that a sub-unit protein vaccine can protect these animals against tuberculosis.  相似文献   

19.
Porcine interleukin-6 gene and CpG sequences were used as immunoadjuvants to enhance the immune responses of newborn piglets to Pseudorabies attenuated vaccine (PAV). The titer of specific antibodies to PAV, the proliferation of lymphocytes and induced IL-2 activities were all examined to identify the immune response of the piglets. The results showed that the immune responses with CpG ODN and porcine interleukin-6 gene were significantly stronger than routine immunities. The data suggests that porcine IL-6 and CpG motifs could be employed as effective immunoadjuvants to raise the humoral and cellular responses of newborn piglets to Pseudorabies attenuated vaccine.  相似文献   

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