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《中国动物传染病学报》2010,(2)
本文综述了目前应用于DNA疫苗的几种免疫佐剂,诸如细胞因子、蛋白质分子、CpG寡核苷酸、化学分子、脂质体等,总结了DNA疫苗免疫佐剂的研究现状,探讨了该领域未来可能的发展方向。 相似文献
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免疫佐剂作用机理的研究进展 总被引：6，自引：1，他引：6
随着DNA重组疫苗、合成肽疫苗等新型疫苗不断涌现,免疫佐剂研究越来越受到人们的关注.尽管佐剂活性物质的研究报道很多,但由于佐剂加强免疫反应是一个非常复杂的过程,关于各种佐剂的作用机制至今尚未完全了解.本文对近年来免疫佐剂作用机理研究进展作-综述,为促进免疫佐剂作用机理的深入研究及免疫佐剂的合理应用提供参考,并初步探讨其作用机理研究的重要性及意义. 相似文献
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从20世纪90年代DNA疫苗出现至今,许多研究者构建了不同种类的DNA疫苗,它具有经济实用、工艺简易及安全高效等特点,因此DNA疫苗已迅速成为疫苗研究领域的热点.这些真核表达载体能够诱发机体产生特异性的体液和细胞介导的免疫应答.对于猪瘟DNA疫苗的研究,国内外研究者做了许多努力并取得了一定的成果.本文将从猪瘟DNA疫苗的构建、免疫佐剂、临床免疫途径以及临床使用的安全性等方面进行阐述. 相似文献
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鸡球虫DNA疫苗研究进展 总被引：1，自引：0，他引：1
鸡球虫病是严重危害养禽业的一种寄生虫病。一直以来,鸡球虫病主要依靠药物来进行防制。但是,随着球虫抗药性问题的日益严重、抗球虫新药开发困难及人们对食品安全的关注,药物在球虫病控制中的作用越来越受到限制,因而人们期望用更好的手段来控制鸡球虫病。DNA疫苗是近年发展起来的一种新型疫苗,因其安全、稳定、高效和易于制备而得到广泛研究。随着鸡球虫保护性抗原基因的不断发现,以及对细胞因子等免疫佐剂在抗球虫感染中的作用不断深入探索,鸡球虫DNA疫苗的研究引起人们的广泛关注和极大兴趣。本文就鸡球虫DNA疫苗作用机理、抗原基因、载体、免疫佐剂、免疫途径、免疫剂量、免疫程序及DNA疫苗研究进展方面展开一系列论述。 相似文献
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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
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为了探索鸡IL-18在禽多杀性巴氏杆菌H基因DNA疫苗中的免疫佐剂作用，分别用共表达鸡IL-18基因和禽多杀性巴氏杆菌C48-1H基因的DNA疫苗、鸡IL-18真核表达质粒pcDNA3／cIL-18与禽多杀性巴氏杆菌C48-1H基因的DNA疫苗混合物、禽多杀性巴氏杆菌C48-1H基因的DNA疫苗肌肉注射5周龄鸡，首免后每周采取外周血及外周抗凝血，应用ELISA和MTT法分别检测免疫鸡的体液免疫及细胞免疫水平。二免后第2周用10 LD50禽多杀性巴氏杆菌强毒菌株C48-1进行攻击。结果鸡IL-18能够明显增强禽多杀性巴氏杆菌H基因DNA疫苗的免疫原性，显著提高免疫鸡的体液免疫和细胞免疫水平，并且鸡IL-18与禽多杀性巴氏杆菌H基因共表达时的免疫佐剂作用最强，能强有力地抵抗强毒菌株C48-1的致死性攻击。结果表明，鸡IL-18可作为DNA疫苗的一种理想的免疫佐剂。 相似文献
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增强DNA疫苗免疫效果的研究进展 总被引：3，自引：1，他引：2
随着DNA重组技术的发展和应用,DNA疫苗、病毒和细菌活载体疫苗等基因工程疫苗的研制日益成为医学领域的一大研究热点。DNA疫苗在许多方面优于传统的灭活苗和减毒苗,但其免疫效果的稳定性和确实性方面尚存不足。影响DNA疫苗免疫效果的因素很多,国内外的许多研究者在这方面作了大量有意义的工作。目前,多数研究者主要通过目的基因的选择、促进外源基因在体内表达、改善疫苗导入方式以及辅以免疫刺激序列和免疫佐剂等方面来提高DNA疫苗的免疫效果。如果DNA疫苗在免疫效果方面能得到大幅度的提高,则它进入临床使用大有前途。 相似文献
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为分离鉴定日本血吸虫含EF手性结构域分子及分析其序列结构特征，首先找到含有EF—hand结构域的分子，按照序列一级结构进行分类，之后进行生物信息学分析。然后用PCR方法以虫卵和成虫cDNA文库为模板扩增分类后的分子，构建重组质粒，诱导蛋白表达并纯化，选取14个纯化蛋白用Western blot进行初步免疫原性鉴定。结果269个原始含EF-hand结构域分子按照序列一级结构分为70个；成功表达和纯化了49个含EF手性分子，进化树分析将其分为9类；Western blot显示，14个纯化蛋白均可被日本血吸虫感染小鼠阳性血清识别。本研究结果为下一步采用这类分子进行动物保护性效果评价以及保护性免疫机制的研究奠定了基础。 相似文献
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蜱的功能分子研究及其应用前景 总被引：29，自引：6，他引：23
蜱既是常见的吸血外寄生虫 ,又是人和动物许多重要疾病的媒介。为了持续地吸血 ,蜱唾液腺可分泌大量生物活性分子 ,并在抗凝血、抗炎症、免疫抑制等方面发挥重要作用。现已分离鉴定出的蜱抗凝血多肽 (TAP)等新型抗凝血和其他多种具有免疫调节的功能分子。此外 ,抗蜱基因工程疫苗候选抗原分子、酶及酶的抑制剂分子、抗药性相关分子、抗菌多肽和毒素等也在被深入研究。这些功能分子不仅在蜱及蜱传染病防制中显示着重要作用 ,同时也在医药生物制剂开发研制上表现出潜在的重要价值 ,显示着它们多方面广阔的应用前景 相似文献
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Evidence is presented to show that activation of endothelial and leucoyte adhesion molecules is a key step in transferring virus from infected leucocytes; and determines the restricted tissue tropism. A range of tissues from 2 experimentally infected mares in late pregnancy at 4 and 8 days after infection with EHV-1 were compared with those from normal pregnant and nonpregnant mares. Rabbit antisera to equine activated endothelial cell molecules were used to identify which tissues expressed these molecules in normal nongravid and gravid mares, and to investigate whether the range of tissues was altered in pregnant mares as a consequence of infection. The results indicated that the endothelium of the pregnant reproductive tract did express these molecules. In the 2 pregnant mares infected with EHV-1, the endothelial cells in the nasal mucosa also expressed these activated endothelial cell molecules. Therefore, the sites expressing these molecules closely correlated with those where virus infection of endothelial cells has been described and is consistent with experimental in vitro data, indicating that expression of these molecules is an essential stage in the transference of virus from leucocytes to endothelial cells. 相似文献
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Persistence of chicken herpesvirus and retroviral chimeric molecules upon in vivo passage 总被引：2，自引：0，他引：2
Mareks disease virus (MDV), a herpesvirus, and avian leucosis virus subgroup J (ALV-J), a retrovirus, were used for experimental coinfection of chickens. Chimeric molecules having sequences of both viruses were detected by the hotspot-combined polymerase chain reaction (HS-cPCR) system. The detection of chimeric molecules provided evidence for avian retroviral inserts in the herpesvirus genome. The persistence of chimeric molecules on in vivo passage served to indicate the infectivity of the recombinant virus. The evaluation of formation and persistence of the chimeric molecules was performed in two trials involving three in vivo passages. The chimeric molecules were identified according to the primer sets, their product length, and pattern. The persistence of chimeric molecules on in vivo passages served as an indication of their ability to replicate in and infect chickens. In the first experimental passage, MDV and ALV-J prototype strains, MD11 and HC-1, were intraperitoneally (i.p.) injected into 1-day-old chicks. The second trial included two passages. Passage II chicks were injected i.p. and passage III chickens were in contact with the chickens of passage II. For passage II, enriched white blood cells from blood samples of chickens from the first trial that had chimeric molecules were injected i.p. into 1-day-old chicks. For passage III, uninfected chicks were included together with the infected chicks. Synthesis evidence for the various species of chimeric molecules was assessed in the tissues of birds of the second trial. DNA was extracted from blood and feathers and analyzed by the hotspot-combined PCR and by pulsed field gel electrophoresis. To overcome the limits of detection, three amplification assays followed by hybridization of the products to specific viral probes were conducted. A variety of chimeric molecules were detected in low concentrations. Five species of chimeric molecules were characterized in blood, tumors, and feathers. Chimeric molecules were detected in 18 of 36 dually infected birds from the first trial and in 14 of 21 dually infected birds from the second trial. The findings show that, in four out of seven groups of the second trial, the chimeric molecule species persisted on passage. 相似文献
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N K Puri K J Gogolin-Ewens M R Brandon 《Veterinary immunology and immunopathology》1987,15(1-2):59-86
The availability of a panel of monoclonal antibodies to sheep MHC class I and class II gene products has allowed for the first time an assessment of the relative complexity of the sheep MHC. By using four monoclonal antibodies to MHC class I, and seven monoclonal antibodies to MHC class II molecules together with one-dimensional SDS-PAGE, sequential immunoprecipitation and 2-dimensional gel analysis, three class I gene products and four distinct subsets of class II molecules have been identified. Sheep class I molecules showed heterogeneity on 2-dimensional gels and as in mouse and man, represented the products of at least three different non-allelic class I genes. Interestingly, the sheep beta 2 microglobulin molecule also displayed heterogeneity, consistent with either two primary gene products or allelic variation. Four sheep class II monoclonal antibodies identified distinct, non-overlapping subsets of sheep class II molecules of Mr 32-36 K (alpha chain) and 25-28 K (beta chain). These class II molecules were co-expressed on sheep B lymphocytes and represented the primary products of different sheep MHC class II genes. The class II molecules within three of these subsets displayed allelic polymorphism essentially restricted to their beta polypeptides, while the fourth subset of class II molecules showed allelic variation in both their alpha and beta polypeptides. The results of this study represent the first evidence for gene duplication and heterogeneity within the sheep MHC. The identification of three primary class I gene products and four distinct subsets of class II molecules suggests three class I loci and up to four distinct class II subregions within the sheep MHC. Potentially large numbers of allelic variants of these different gene products may be expressed in normal sheep. 相似文献
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The presentation of antigen to specific T-cell populations is a crucial event during the elicitation phase of contact hypersensitivity (CHS). Significant changes in CD4(+) T-cell and gammadelta T-cell populations occur in the skin of sheep 48h after re-exposure to dinitrochlorobenzene but the expression of antigen presentation molecules such as MHC-II and CD1 at this stage of the hypersensitivity response has not been investigated. In the present study, a panel of monoclonal antibodies recognising CD1 and MHC-II subtypes was used in combination with computer assisted morphometric analysis to estimate the distribution of antigen presentation molecules in the superficial and deep dermis of the ears of lambs during the elicitation phase of CHS. The MHC-II molecules showed predominantly a perivascular and peri-appendageal distribution in the dermis and there were scattered MHC-II(+) cells in the basal and suprabasal layers of the epidermis. The CD1w2(+) (CD1b-like) molecules were present on distinct cells that were scattered evenly through the dermis, whereas CD1w3(+) (CD1c-like) molecules were almost exclusively detected on or in close association with the vascular endothelium. There was a significant increase in the presence of MHC-DQ(+) cells in the superficial dermis of dinitrochlorobenzene-treated animals compared with both an untreated control group and a vehicle-treated control group. However, MHC-DQ/DR(+) and CD1w3(+) cells only showed a significant increase compared with the vehicle-treated control group. The present study shows that the distribution of molecules involved in antigen presentation to CD4(+) T-cells and gammadelta T-cells changes during the elicitation phase of CHS in sheep, and suggests a role for MHC-DQ molecules on antigen presenting cells. However, the changes in distribution and expression of MHC-II and CD1 subtypes argue against a prominent role for a CD1-dependent pathway for T-cell recognition in the clinical cutaneous hypersensitivity response in sheep. Based on the expression of MHC-II molecules and CD1c molecules, we also suggest a potential role for endothelial cells in antigen presentation during the clinical dermatitis reaction. 相似文献
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OBJECTIVE: To describe the microanatomic features of pancreatic islets and the immunohistochemical distribution of glucose transporter (GLUT) molecules in the pancreas and other tissues of New World camelids. ANIMALS: 7 healthy adult New World camelids, 2 neonatal camelids with developmental skeletal abnormalities, and 2 BALB/c mice. PROCEDURE: Samples of pancreas, liver, skeletal muscle, mammary gland, brain, and adipose tissue were collected postmortem from camelids and mice. Pancreatic tissue sections from camelids were assessed microscopically. Sections of all tissues from camelids and mice (positive control specimens) were examined after staining with antibodies against GLUT-1, -2, -3, and -4 molecules. RESULTS: In camelids, pancreatic islets were prominent and lacked connective tissue capsules. Numerous individual endocrine-type cells were visible distant from the islets. Findings in neonatal and adult tissues were similar; however, the former appeared to have more non-islet-associated endocrine cells. Via immunostaining, GLUT-2 molecules were detected on pancreatic endocrine cells and hepatocytes in camelids, GLUT-1 molecules were detected on the capillary endothelium of the CNS, GLUT-3 molecules were detected throughout the gray matter, and GLUT-4 molecules were not detected in any camelid tissues. Staining characteristics of neonatal and adult tissues were similar. CONCLUSIONS AND CLINICAL RELEVANCE: In New World camelids, microanatomic features of pancreatic islets are similar to those of other mammals. Data suggest that the poor glucose clearance and poor insulin response to hyperglycemia in adult camelids cannot be attributed to a lack of islet cells or lack of GLUT molecules on the outer membrane of those cells. 相似文献
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The anti-CD1 monoclonal antibodies submitted to the 1st International Workshop on Leucocyte Differentiation Antigens of Cattle, Sheep and Goats were tested for their reactivity on sheep skin, thymus and lymph node and for their reactivity with sheep efferent and afferent lymph and peripheral blood. With the exception of 20-27 they all stained that same cell populations. The antibodies precipitated molecules with a heavy chain of 46,000 apparent molecular weight and a light chain of 14,000 apparent molecular weight. VPM5 and CC14 antigens were purified by affinity chromatography. All the antibodies cross-reacted with these molecules. The results show that 20-27 recognises the same molecules as the other antibodies and suggest that 20-27 is a pan CD1 monoclonal antibody and the other monoclonal antibodies are homologues of the human CD1b molecules. 相似文献
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Buchanan MF Carter WC Cowgill LM Hurley DJ Lewis SJ MacLeod JN Melton TR Moore JN Pessah I Roberson M Robertson TP Smith ML Vandenplas ML 《Journal of veterinary medical education》2005,32(1):72-78
Traditional methods of teaching intracellular biological processes and pathways use figures or flowcharts with the names of molecules linked with arrows. Many veterinary students, presented with such material, simply memorize the names or chemical structures of the molecules and are then likely to forget the material once the examination is completed. To address this problem, the authors designed, created, and field-tested new teaching media that incorporate realistic three-dimensional (3D) animations depicting the dynamic changes that occur in intracellular molecules during cellular activation. Testing found that veterinary students taught using traditional teaching media (e.g., lectures, handouts, textbooks) are proficient in memorizing the names and order of intracellular molecules but unable to appreciate the interactions between these elements or their spatial relationships within cells. In contrast, more than 90% of veterinary students taught using 3D animations not only recall the facts about the intracellular elements but also develop accurate mental images of the interactions among these molecules and their spatial relationships. These findings strongly suggest that the comprehension of complex biological processes by veterinary students can be enhanced by the use of dynamic 3D depictions of these processes in the classroom. 相似文献