CpG ODN序列结构特征对免疫活性影响的研究进展

CpG ODN(CpG oligonucleotide,CpG 寡脱氧核苷酸)是人工合成的含有非甲基化的胞嘧啶鸟嘌呤二核苷酸(CpG)的寡脱氧核苷酸(ODN),可模拟细菌DNA刺激多种哺乳动物包括人的免疫细胞.     

免疫调节性寡聚脱氧核苷酸筛选方法的建立、优化和应用

目的:建立一套合理而便捷的实验体系,为开发新型免疫调节性寡聚脱氧核苷酸(ODN)提供筛选方法。方法:利用含有CpG基序的免疫刺激性寡聚脱氧核苷酸(CpG ODN)作为刺激物,将人外周血单个核细胞(PBMC)作为研究对象,分别以细胞的增殖程度和刺激上清的抗病毒活性作为检测指标,优化各项实验条件,综合评定寡聚脱氧核苷酸的免疫调节活性。结果:成功建立了由阳性免疫调节性ODNA151抑制CpG ODN诱导的人PBMC增殖及抗病毒活性的筛选方法。结论:免疫调节性ODN筛选方法的成功建立,为下一步研究开发新型免疫调节性ODN奠定了基础。     

DNA免疫诱导小鼠产生抗人心肌肌钙蛋白T抗血清的研究

目的 通过基因免疫诱导BALB／c小鼠产生抗人心肌肌钙蛋白T(cTnT)的抗体应答，并比较不同免疫佐剂、预处理方法对免疫应答的影响。方法 构建cTnT的真核表达质粒pCI-cTnT，大量提取、PEG纯化质粒，通过肌肉注射免疫BALB／c小鼠并加强免疫，每隔2周取血，ELISA法分析抗cTnT的体液应答，免疫印迹检测抗血清的特异性。并比较布吡卡因预处理、CpG ODN核酸佐剂、弗氏不完全佐剂对抗体应答效果的影响。结果 接种pCI-cTnT质粒的小鼠血清中检测出抗cTnT特异性抗体，加强免疫使抗体滴度提高；使用CpGODN后免疫应答增强，弗氏不完全佐剂可明显增强免疫效果，而吡卡因预处理使应答水平下降；初次免疫后14周仍可测得较高的抗体应答。结论 采用含cTnT基因的质粒DNA免疫，成功地诱导小鼠产生抗cTnT的抗体；传统的弗氏不完全佐剂可作为核酸免疫的佐剂使用。     

Anti-proliferative effects of phosphodiester oligodeoxynucleotides

The immunostimulatory effects of cytosine-phosphate-guanosine (CpG)-containing oligodeoxynucleotides (ODNs) have been extensively documented. In this paper, we describe the inhibitory effects of ODNs that contain natural phosphodiester backbones (O-ODNs) on the immunostimulation caused by CpG-containing phosphorothioated ODNs (CpG-S). CpG-S stimulation of mouse splenocyte proliferation was reduced by the addition of O-ODNs that contained or lacked the CpG-motif (CpG-containing phosphodiester oligodeoxynucleotide, CpG-O or GpC-O). The total number of cultured splenocytes was up-regulated by CpG-S, whereas repetitive addition of O-ODNs to the cell cultures inhibited this effect. The frequency of T2-like B cells was found to be increased by CpG-S. The culture supernatants of CpG-S-treated splenocytes contained elevated levels of IL-10 and IL-6. However, IL-10 and IL-6 production was down-regulated significantly by the combination of CpG-S and either CpG-O or GpC-O. The O-ODN mediated inhibition of proliferation was less pronounced in IL-10-/- mice. Thus, the O-ODNs, irrespective of CpG content, exerted inhibitory activities on the proliferation of B cells. These anti-proliferative effects appear to be mediated both by the down-regulation of IL-10 production and increased apoptosis.     

含CpG寡聚脱氧核苷酸对肿瘤抗原肽P815AB的免疫佐剂效应研究

目的 以肿瘤抗原Ｐ８１５ＡＢ抗原肽为模式抗原,研究ＣｐＧ寡聚脱氧核苷酸（ＣｐＧ ＯＤＮ）对该肿瘤抗原表位肽的免疫佐剂效应。方法 用［＾３Ｈ］－ＴｄＲ掺入法研究ＣｐＧ ＯＤＮ体外刺激小鼠脾细胞增殖效应,用ＥＬＩＳＰＯＴ和［＾３Ｈ］－ＴｄＲ释放法检测不同构成形式的疫苗在体诱发特异性ＣＴＬ应答的效应。结果 ＯＤＮ１８２６在体外可刺激小鼠脾细胞显著增殖,经４次免疫,在“肽＋ＯＤＮ１８２６＋ＩＦＡ”组中,４     

不同剂量CpG ODN联合STAg鼻内免疫BALB／c小鼠诱导的免疫应答

目的 观察弓形虫可溶性速殖子抗原（STAg）与不同剂量的CpG ODN联合滴鼻免疫小鼠的免疫效果，探索CpG ODN佐剂最佳剂量。方法 5-6周龄BALB／c小鼠50只随机分为5组，每组10只，实验组以20μg STAg联合不同剂量CpG ODN佐剂滴鼻免疫小鼠，佐剂剂量分别为0、1、5、10μg，对照组以PBS滴鼻，间隔2wk免疫2次。末次免疫后30d断颈处死全部小鼠，分离脾、Peyer＇s patch（PP结）、肠系膜淋巴结（MLN）和IEL淋巴细胞，观察淋巴细胞增殖情况。用ELISA的方法检测血清IgG和粪便IgA含量。结果 与PBS组和0剂量佐剂组相比，10μg CpG ODN联合STAg免疫后，小鼠肠粘膜淋巴组织和脾淋巴细胞增殖明显（P〈0．05）。10μg组血清IgG远高于对照组（P〈0．05），粪便IgA高于对照组（P〉0．05）。结论 CpG ODN可作为STAg的粘膜佐剂，10μg CpG ODN与STAg联合具有最佳的免疫效果。     

�Ѻ�����MT01�����������׷�����о���չ

??MT01 is a single-stranded oligodeoxynucleotide??ODN??which is designed based on human mitochondrial DNA sequences. It has immune activity of selective negative regulator. Several studies have shown that MT01 has a significant effect in suppressing excessive immune reaction caused by pathogens infection??and it also inhibits the absorption of alveolar bone and promotes osteoblast cell activation. Inflammation and bone resorption are the nature of periodontal diseases. Here we review the latest research progress of MT01 and its related research in periodontitis.     

Delivery of an inactivated avian influenza virus vaccine adjuvanted with poly(D,L-lactic-co-glycolic acid) encapsulated CpG ODN induces protective immune responses in chickens

In poultry, systemic administration of commercial vaccines consisting of inactivated avian influenza virus (AIV) requires the simultaneous delivery of an adjuvant (water-in-oil emulsion). These vaccines are often limited in their ability to induce quantitatively better local (mucosal) antibody responses capable of curtailing virus shedding. Therefore, more efficacious adjuvants with the ability to provide enhanced immunogenicity and protective anti-AIV immunity in chickens are needed. While the Toll-like receptor (TLR) 21 agonist, CpG oligodeoxynucleotides (ODNs) has been recognized as a potential vaccine adjuvant in chickens, poly(D,L-lactic-co-glycolic acid) (PLGA) nanoparticles, successfully tested as vaccine delivery systems in other species, have not been extensively explored. The present study, therefore, assessed both systemic and mucosal antibody-mediated responses following intramuscular vaccination (administered at 7 and 21 days post-hatch) of chickens with PLGA encapsulated H9N2 AIV plus encapsulated CpG ODN 2007 (CpG 2007), and nonencapsulated AIV plus PLGA encapsulated CpG 2007 vaccine formulations. Virus challenge was performed at 2 weeks post-secondary vaccination using the oculo-nasal route. Our results showed that chickens vaccinated with the nonencapsulated AIV vaccine plus PLGA encapsulated CpG 2007 developed significantly higher systemic IgY and local (mucosal) IgY antibodies as well as haemagglutination inhibition antibody titres compared to PLGA encapsulated AIV plus encapsulated CpG 2007 vaccinated chickens. Furthermore, chickens that received CpG 2007 as an adjuvant in the vaccine formulation had antibodies exhibiting higher avidity indicating that the TLR21-mediated pathway may enhance antibody affinity maturation qualitatively. Collectively, our data indicate that vaccination of chickens with nonencapsulated AIV plus PLGA encapsulated CpG 2007 results in qualitatively and quantitatively augmented antibody responses leading to a reduction in virus shedding compared to the encapsulated AIV plus PLGA encapsulated CpG 2007 formulation.     

Randomized,double-blind,placebo-controlled,safety and immunogenicity study of 4 formulations of Anthrax Vaccine Adsorbed plus CPG 7909 (AV7909) in healthy adult volunteers

A new anthrax vaccine that could accelerate the immune response and possibly reduce the number of injections needed for protection would be desirable in a post-exposure setting.