题目名称：（英文） （中文） 体结合微粒的作用
题目名称：（英文）
的定植性和免疫原性
题目名称：（中文) 基因真核表达质粒的
（英文）
学生姓名： 学 号：
院系专业：教学班级：
硕士导师： 职 称： 博 导
研究方向：
2014年 1 月 5 日
The role of antigen specificity in the binding of murine monoclonal anti-DNA antibodies to microparticles from apoptotic cells
Name： Major：Preventive Veterinary Medicine
Tutor：
Abstract：Antibodies to DNA (anti-DNA) are the serological hallmark of systemic lupus erythematosus and markers of underlying immune system disturbances. These antibodies bind to both single-stranded and double-stranded DNA, mediating pathogenesis by forming immune complexes. As shown recently, DNA in blood exists in both free and particulate forms, with DNA representing an important component of microparticles. Microparticles are membrane-bound vesicles containing nuclear molecules, released by membrane blebbing during cell death and activation. A panel of monoclonal NZB/NZW F1 anti-DNA antibodies was tested for binding to microparticles generated from apoptotic THP-1 and Jurkat cells. These studies showed that only certain anti-DNA antibodies in the panel, specific for double-stranded DNA, bound to microparticles. Binding to particles was reduced by soluble DNA or DNase treatment. Together, these results indicate that particle binding is a feature of only certain anti-DNA antibodies, reflecting immunochemical properties of the antibodies and the nature of the exposed DNA antigens.
Keywords: Children Mercury Sleep Cytokines IL-6 TNF-α
从凋亡细胞抗原特异性与鼠单克隆抗DNA抗体结合微粒的作用
姓 名：
摘 要：抗体的DNA（抗DNA）是系统性红斑狼疮和底层免疫系统紊乱标志物的血清学标志。这些抗体结合至两个单链和双链DNA，通过形成免疫介导的发病机制复合物。如最近表明，在血液中的DNA以游离和颗粒形式存在，是DNA代表微粒的一个重要组成部分。微粒含有核分子膜结合的囊泡，通过细胞死亡释放的膜起泡激活。单克隆NZB/ NZW的F1抗DNA抗体的面板被结合到从凋亡的THP-1和Jurkat细胞中产生的微粒进行测试。这些研究显示，在面板只有某些抗DNA抗体，特异于双链DNA，结合到微粒。结合到粒子减少了可溶性DNA或DNA酶处理。总之，这些结果表明，颗粒的结合是只有某些抗DNA抗体的特征，反映了抗体的免疫化学性质和DNA的暴露抗原的性质。
关键词：微泡;抗DNA;狼疮;自身免疫;细胞凋亡
Oral administration of live Shigella vaccine candidates in rhesus
monkeys immunogenicity between different serotypes
Name：
Abstract: Live oral monovalent Shigella flexneri 2a vaccine candidates as well as bivalent formulations with Shigella sonnei were evaluated in a rhesus monkey model for colonization and immunogenicity. Freshly harvested suspensions of S. flexneri 2a vaccine candidates WRSf2G12 and WRSf2G15 as well as S. sonnei accine candidate WRSs3 were nasogastrically administered to groups of rhesus monkeys, Macaca mulatta, either in a monovalent form or when combined with each other. The animals were monitored daily for
physical well-being, stools were subjected to quantitative colony immunoblot assays for bacterial excretion and blood and stools were evaluated for humoral and mucosal immune responses .No clinical symptoms were noted in any group of animals and the vaccine candidates were excreted ro bustly for 48–72 h without significant changes in either the magnitude or duration of excretion when given as a monovalent or as bivalent mixtures.Similarly, immunological interferences were not apparent in the magnitude of humoral and mucosal immune responses observed toward Shigella-specific antigens when monkeys were fed monovalent or bivalent formulations. These results predict that a multivalent live oral vaccine of more than one serotype can have a favorable outcome for protection against shigellosis.
Key words: Shigella Live bivalent vaccine candidates Rhesus monkeys Colonization Immunogenicity
活志贺氏菌候选口服疫苗在恒河猴的不同血清型之间的免疫原性
姓 名
摘 要：痢疾杆菌口服活价疫苗选定，以及二价的制剂与志贺氏菌在恒河猴进行评价模型免疫原性定植。新鲜收获的悬浮液福氏2a中的候选疫苗WRSf2G12和WRSf2G15宋氏志贺菌疫苗候选人WRSs3被给予恒河猴群体猴子，猕猴，无论以单价形式，或当彼此结合。动物每天监测身体健康，粪便经受定量菌落免疫印迹法进行细菌排泄和血液和粪便进行了体液免疫和粘膜免疫应答的评价无临床症状，指出在任何一组的动物，当作为一价或二价作为混合.同样给出的候选疫苗进行了排泄ロ催化48-721h而不需在任何大小或排泄的持续时间显著变化，免疫干扰不明显的体液和粘膜朝向时猴子喂食的一价或二价配制剂志贺氏菌特异性抗原观察到免疫应答的幅度。这些结果预测，一个以上的血清型的多价活口服疫苗可以具有用于防止细菌性痢疾一个有利的结果
关键词：志贺氏 价活疫苗候选 河猴的免疫原性定植
犬细小病毒HL- 01 株的分离鉴定及VP2基因真核表达质粒的构建 姓 名
作者：王玉玲
单位：东北农业大学
摘 要：从临床发病犬采集粪便样品, 以F81 传代细胞进行病毒分离, 经血球凝集实验( HA) 和血球凝集抑制实验( HI) 初步鉴定为犬细小病毒。为进一步确诊, 根据Genbank 中已发表的犬细小病毒VP2 基因序列设计并合成一对引物, 通过聚合酶链反应( PCR) 扩增出CPV VP2 基因, 酶切, 测序加以鉴定。其序列与国际已发表的CPV- d( type
2) 、CPV- 15( type 2a) 、CPV- 39( type 2b) VP2 序列同源性分别为98.97%, 98.75%, 98.69%, 氨基酸序列的同源性分别为98.12%, 97.60%, 97.60%, 从而证明此分离株为犬细小病毒。在获得VP2 基因的基础上, 为实现VP2蛋白的表达, 构建了真核表达质粒pMel BacC- VP2。
关键词：犬细小病; VP2 基因; 克隆; 表达质粒; 构建
construction of eukaryotic expr ession plasmid of VP2 gene and
Identification of HL- 1 isolate of canine parvovirus
Name：
Abstract:Identification of canine parvovirus.from clinical onset of canine collecting fecal samples of virus isolation, to F81 cells, the cells agglutination test (HA) and hemagglutination inhibition (HI) . For further diagnosis, according to canine parvovirus VP2 gene sequence published in Genbank were designed and synthesized a pair of primers, polymerase chain reaction (PCR) amplified CPV VP2 gene, enzyme digestion, sequencing identified.The sequence of published CPV- D (type 2), CPV- 15 (type 2A), CPV- 39 (type 2B) VP2 sequence homology were 98.97%, 98.75%, 98.69%, the amino acid sequence homology were 98.12%, 97.60%, 97.60%, and thereby prove that the isolates of canine parvovirus. On the base of VP2 gene, expression of VP2 protein in order to achieve, to construct the eukaryotic expression plasmid pMel BacC- VP2.
Key words: Canine parvovirus disease; VP2 gene; cloning; expression plasmid; construction
References
[1] http://dx.doi.org/10.1016/j.clim.2014.05.007 a Duke University Medical Center, Department of Medicine, Durham, NC 27710, USA b Department of Microbiology, Immunology and Biochemistry, The University of Tennessee Health Science Center, Memphis,TN 38163, USA c Medical Research Service, Durham Veterans Administration Medical Center, Durham, NC 27705, USA
[2] http://dx.doi.org/10.1016/j.vaccine.2013.12.068 A Bacterial Diseases Branch,Walter Reed Army Institute of Research,503 Robert Grant Avenue,Silver Spring,MD 20910,United States
B Diseases Branch, Naval Medical Research Center (NMRC),503 Robert Grant Avenue,SilverSpring,MD20910,United States C department of Veterinary Medicine, Walter Reed Army Institute of Research (WRAIR), Silver Spring, MD, United State
[3]
Appendix I
Identification of HL- 1 isolate of canine parvovirus
and construction of eukaryotic expr ession plasmid of VP2 gene Name
Abstract: The HL- 01 isolate of canine parvovirus was propagated on F81 cell. And identified by the haem agglutination test (HA) and haem agglutination inhibition test (HI). Using a pair of primers based on the published sequence of CPV′s VP2 gene, the full length gene encoding outer capsid protein VP2 was amplified from cell culture by polymerase chain reaction (PCR). The VP2 gene of HL- 01 isolate of canine parvovirus was inserted into pMD18- T vector and was identified by sequencing, digestion. The result showed that there was a high homology innucleotide sequence and aminoacid sequence incomparison with CPV- d (type 2), CPV- 15(type 2a) and CPV- 39 (type 2b). The rate of nucleotide sequence homology was 98.97%, 98.75%, 98.69%, and the aminoacid homology was 98.12%, 97.60%, 97.60%. The VP2 gene of the isolate was inserted into the eukaryotic expression plasmid pMel BacC and construced recombinated plasmid pMel BacC- VP2 in order to realize the expression of VP2 protein.
Key words: canine parvovirus; VP2; cloning; pMel BacC expression plasmid; construction