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1.
为探究硫酸软骨素N-乙酰半乳糖胺基转移酶-1(chondroitin sulfate N-acetylgalactosaminyltransferase-1,CSGalNAcT-1)在克氏原螯虾免疫反应中的作用,利用RACE技术成功获得克氏原螯虾CSGalNAcT-1基因完整cDNA序列,并通过荧光定量PCR分析该基因在各组织中的表达模式以及在CpG寡脱氧核苷酸(ODN)、嗜水气单胞菌和白斑综合征病毒(WSSV)刺激后的表达情况。结果显示,克氏原螯虾CSGalNAcT-1基因在中肠的相对表达量最高,其次是近胃段肠,在心脏、胃、肌肉和围食道神经中的相对表达量较低。克氏原螯虾肝胰腺、血细胞中CSGalNAcT-1基因表达量在嗜水气单胞菌和WSSV刺激下呈现出下调及上调,而在注射免疫刺激剂CpG ODN后,CSGalNAcT-1基因表达水平在大部分组织中呈现上调。表明克氏原螯虾CSGalNAcT-1基因可能在抵御外界病原侵染以及增强自身免疫反应的过程中发挥重要作用。  相似文献   
2.
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   
3.
用添加CpG寡聚核苷酸(CpG ODN)和表面展示VP28的解脂耶罗维亚酵母(VP28-yl)的饵料投喂凡纳滨对虾,进行田间中试实验.投喂30 d后进行WSSV感染实验,评估其对凡纳滨对虾的免疫保护作用.投喂实验结束后,CpG ODN投喂组对虾的相对增重率达到(65.8±7.8)% (P<0.05),这暗示CpG ODN可能具有促生长作用.WSSV攻毒后,CpG ODN和VP28-yl投喂组对虾中WSSV拷贝数与对照组相比均显著降低(P<0.05),相对免疫保护率分别可达到 26.7%和 36.7%.在投喂结束和WSSV刺激后,CpG ODN组对虾中的呼吸爆发水平均显著升高(P<0.05).而在VP28-yl投喂组,WSSV引起的细胞凋亡则显著受到抑制(P<0.05).此外,WSSV刺激后,STAT基因在CpG ODN组和VP28-yl组对虾中的表达水平均显著上调(P<0.05),分别在第5天和第3天达到最大值,而对照组中则显著下调.研究结果表明,CpG ODN和VP28-yl增强了凡纳滨对虾抗病毒免疫力,对养殖对虾病毒性疫病的防控具有显著作用,可以作为免疫增强剂添加在饵料中,具有在养殖生产中推广使用的前景.  相似文献   
4.
Immunostimulatory sequences of oligodeoxynucleotides (ODNs), such as CpG ODNs, are potent stimulators of innate immunity. Here, we identified a strong immunostimulatory CpG ODN, which we named MsST, from the lac Z gene of Streptococcus (S.) thermophilus ATCC19258, and we evaluated its immune functions. In in vitro studies, MsST had a similar ability as the murine prototype CpG ODN 1555 to induce inflammatory cytokine production and cell proliferation. In mouse splenocytes, MsST increased the number of CD80+CD11c+and CD86+CD11c+ dendritic cells and CD4+CD25+ regulatory T cells. We also analyzed the effects of MsST on the expression of regulatory cytokines by real-time quantitative PCR. MsST was more potent at inducing interleukin-10 expression than the ODN control 1612, indicating that MsST can augment the regulatory T cell response via Toll-like receptor 9, which plays an important role in suppressing T helper type 2 responses. These results suggest that S. thermophilus , whose genes include a strong Immunostimulatory sequence-ODN, is a good candidate for a starter culture to develop new physiologically functional foods and feeds.  相似文献   
5.
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   
6.
为获得对猪具有免疫刺激活性的含CpG序列的重组质粒,设计合成了11条CpG序列,应用MTS比色法测定合成CpG ODNs刺激猪PBMC增殖的能力,结果有10条CpG ODN对猪PBMC具有刺激活性(SI>2);以对猪PBMC具有较高刺激活性的CpG06和CpG08为核心,分别合成含6次重复序列的重组质粒pCpG06和pCpG08,并检测其对猪PBMC的刺激活性,结果重组质粒pCpG06和pCpG08对猪PBMC的刺激指数分别可达4.97和5.32,并可刺激猪PBMC产生较高水平的IL-4。研究获得的重组质粒pCpG06和pCpG08对猪PBMC具有较好的刺激活性,可为猪用CpG佐剂的研究提供参考。  相似文献   
7.
We developed a severe anaphylactic model in mice using buckwheat antigen and B‐type CpG‐oligodeoxynucleotides (CpG‐ODNs) from Streptococcus thermophilus genome. In typical systemic anaphylaxis models, animals are challenged with large quantity of antigens via an intravenous (i.v.) route. Here, we showed a simple anaphylactic shock after challenge via intraperitoneal (i.p.) route. The i.p. method is simpler than i.v. administration and has a lower risk for failure. To generate this anaphylactic model, 5‐week‐old female BALB/c mice were first i.p. sensitized with buckwheat antigen mixed with B‐type CpG‐ODN. After 2 weeks, mice were challenged with antigen to induce anaphylactic shock, which was evaluated by scoring the severity symptoms and measuring serum levels of various proteins and splenic cell producing cytokines. Immunoglobulin (Ig)G2a production and interferon‐γ positive cells were markedly increased in mice immunized with antigen mixed with B‐type CpG‐ODN, whereas serum IgE levels were decreased by B‐type CpG‐ODN. We also examined the effects of various ODNs (A, B and C‐type CpG‐ODNs) and antigens (buckwheat, α‐casein, β‐lactoglobulin and ovalbumin) on anaphylactic severity, and found that the combination of buckwheat and B‐type CpG‐ODN induced the most intense anaphylactic shock. This model is expected to contribute to the study of the prevention of anaphylactic shock.  相似文献   
8.
CpG ODN对鸡新城疫LaSota活疫苗的免疫增强效应   总被引:4,自引:0,他引:4  
将3种不同的未甲基化CpGODN分别与新城疫LaSota活疫苗混合后,经滴鼻和点眼免疫鸡,通过检测鸡血清中HI抗体、外周血T淋巴细胞增殖活性、诱导巨噬细胞分泌NO含量,以及刺激外周血淋巴细胞表达IFN-γ、IL-6与IL-1β mRNA量,分析各CpGODN对新城疫LaSota活疫苗免疫效果的影响。结果表明,经2次免疫后,含GTCGTT核心基序的CpG ODN1组,鸡血清平均HI抗体效价最高达8.2log2、淋巴细胞刺激指数达9.836、NO分泌量达35.833μmol/L,分别比疫苗单独免疫组高出2个滴度(P〈0.05)、4.4(P〈0.01)、27.6μmol/L(P〈0.01);含GACGTT核心基序的CpG ODN2组增强作用不明显.与疫苗单独免疫组无差异;而CpGODN3的免疫刺激活性由于受其侧翼序列的影响,作用明显减弱甚至丧失。对细胞因子的影响,CpGODN3组IFN-γ mRNA表达量稍高于CpG ODN1组,而其余细胞因子均以CpG ODN1组表达水平最高(P〈0.05)。由此证明CpG ODN1能显著增强鸡对新城疫LaSota活疫苗的体液和细胞免疫反应,可以作为高效的免疫增强剂。  相似文献   
9.
为获得对水貂具有免疫刺激活性的CpG ODN序列,设计合成了45种含不同CpG基序的DNA序列,应用MTS比色法测定合成CpG ODNs刺激水貂PBMC增殖的能力,结果有11条CpG ODN对水貂PBMC有刺激活性(SI>2);应用SI值大于5的6种CpG ODN分别与水貂伪狂犬灭活疫苗联合免疫水貂,免疫后经水貂血清抗伪狂犬中和抗体效价测定和水貂PBMC非特异性增殖效应检测,结果有3个CpG ODN序列对水貂具有较好免疫增强作用,分别是CpG-21(ATCGATTTGTCGTTATCGAT)、CpG-23(ATCGATGAACGTTAACGTTTC)和CpG-24(AACGTTCATCGATATCGATGT)。本研究获得3条对水貂具有较好免疫刺激活性的CpG ODN序列,可为水貂新型CpG佐剂的研究提供参考。  相似文献   
10.
用添加CpG寡聚核苷酸(CpG ODN)和表面展示VP28的解脂耶罗维亚酵母(VP28-yl)的饵料投喂凡纳滨对虾,进行田间中试实验。投喂30 d后进行WSSV感染实验,评估其对凡纳滨对虾的免疫保护作用。投喂实验结束后,CpG ODN投喂组对虾的相对增重率达到(65.8±7.8)% (P<0.05),这暗示CpG ODN可能具有促生长作用。WSSV攻毒后,CpG ODN和VP28-yl投喂组对虾中WSSV拷贝数与对照组相比均显著降低(P<0.05),相对免疫保护率分别可达到26.7%和36.7%。在投喂结束和WSSV刺激后,CpG ODN组对虾中的呼吸爆发水平均显著升高(P<0.05)。而在VP28-yl投喂组,WSSV引起的细胞凋亡则显著受到抑制(P<0.05)。此外,WSSV刺激后,STAT基因在CpG ODN组和VP28-yl组对虾中的表达水平均显著上调(P<0.05),分别在第5天和第3天达到最大值,而对照组中则显著下调。研究结果表明,CpG ODN和VP28-yl增强了凡纳滨对虾抗病毒免疫力,对养殖对虾病毒性疫病的防控具有显著作用,可以作为免疫增强剂添加在饵料中,具有在养殖生产中推广使用的前景。  相似文献   
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