日本七鳃鳗免疫方案的建立及抗血清杀伤活性的鉴定

发布时间：2018-06-11 11:28

本文选题：日本七鳃鳗 + 可变淋巴细胞受体　；参考：《辽宁师范大学》2012年硕士论文

【摘要】：在针对适应性免疫系统起源与进化的探究中,现存的无颌类脊椎动物的代表一七鳃鳗因其独特的进化地位及特有的适应性免疫系统从而受到了免疫学家的高度关注,继而成为这一研究领域中的新型模式生物。随着生物系统发生的进行,以哺乳动物为代表的有颌类脊椎动物通过天然免疫系统和适应性免疫系统的协同作用来完成对外界抗原刺激的免疫应答。其中以免疫球蛋白(Immunoglobulin, Ig)、T/B淋巴细胞表面受体(TCR/BCR)和主要组织相容性复合物(MHC)为代表免疫球蛋白超家族(Immunoglobulin superfamily,IgSF)成员,赋予了适应性免疫应答所特有的复杂性、专一性和记忆性。 2004年Pancer Z等学者经研究证实,在无颌类脊椎动物七鳃鳗中存在一种独特的、由可变淋巴细胞受体(variable lymphocyte receptors,VLRs)介导的适应性免疫系统。目前已经证实在七鳃鳗体内存在三类VLR分子—VLRA、VLRB及VLRC;当机体受到外源微生物刺激后,只有VLRB＋淋巴细胞会发生增殖并分化成浆细胞,分泌可溶性VLRB分子参与免疫应答。由此可见,七鳃鳗拥有独特的、唯一一种不依赖于免疫球蛋白的适应性免疫体系,对该免疫系统作用机理的的探究,将为解开适应性免疫系统起源及进化这一由来已久的谜团提供关键线索。遗憾的是,目前在七鳃鳗免疫研究领域仍缺少一套标准化的免疫方案,不仅造成了七鳃鳗免疫流程中抗原制备、免疫方式、免疫时间等相关因素的不确定性,更阻碍了后续抗血清活性及VLR免疫应答体系的研究。为解决这一问题,本研究运用多种抗原免疫日本七鳃鳗(Lampetra japonica),分别为大肠杆菌(Escherichia coli, E. coli),变形杆菌(Bacillus proteus, B. proteus),耻垢分枝杆菌(Mycobacterium smegmatis, M. smegmatis)金黄色葡萄球菌(Staphylococcus aureus, S. aureus);兔红细胞(RRBCs),绵羊红细胞(SRBCs);肿瘤细胞NB4、HepG2及Hela。采取腹腔注射及肌肉注射两种免疫途径,每十天进行一次免疫,共免疫4次,随后采集七鳃鳗抗血清。此外,作者对七鳃鳗抗血清的杀菌及溶细胞活性进行了进一步的探究,为针对七鳃鳗体内VLR介导的适应性免疫系统的研究提供基础,从而进一步推动脊椎动物适应性免疫系统起源及进化的相关研究。
[Abstract]:In the exploration of the origin and evolution of adaptive immune system, the representative of the extant edentulous vertebrate, lamprey, has been highly concerned by immunologists for its unique evolutionary status and unique adaptive immune system. Then it becomes a new model organism in this research field. With the development of biological system, the vertebrate of jaw represented by mammal can complete the immune response to the stimulation of external antigen by the synergistic action of innate immune system and adaptive immune system. Immunoglobulins, T / B lymphocyte surface receptors (TCR / BCRs) and major histocompatibility complexes (MHCs) were used as members of immunoglobulin superfamily IgSFs, which gave the specific complexity of adaptive immune response. Specificity and memory. In 2004, Pancer Z and other researchers confirmed that there is a unique adaptive immune system mediated by variable lymphocyte receptor (lymphocyte receptor) in lamprey, a non-maxillary vertebrate. At present, it has been proved that there are three kinds of VLR molecules-VLRAN VLRB and VLRC in lamprey. When the organism is stimulated by exogenous microorganisms, only VLRB+ lymphocytes will proliferate and differentiate into plasma cells and secrete soluble VLRB molecules to participate in the immune response. The lamprey has a unique, only adaptive immune system independent of immunoglobulin, which explores the mechanism of its action. It will provide a key clue to unravel the long-standing mystery of the origin and evolution of the adaptive immune system. Unfortunately, there is still a lack of a standardized immunization program in the field of lamprey immunization, which not only leads to the uncertainty of antigen preparation, immunization methods, immune time and other related factors in the immune process of lamprey. It also hinders the further study of antiserum activity and VLR immune response system. In order to solve this problem, In this study, Lampetra japonica, Escherichia coli, E. coli, Bacillus proteus, B. proteusus, Mycobacterium smeatisgmatism, Mycobacterium smegmatisme, Staphylococcus aureus, Saureusus, RRBCsof rabbit erythrocytes, SRBCsof sheep, tumor cells NB4HepG2 and Hela. were immunized with various antigens. The mice were immunized once every ten days by intraperitoneal injection and intramuscular injection. In addition, the authors further explored the bactericidal and cytolytic activity of lamprey antiserum, which provided a basis for the study of adaptive immune system mediated by VLR in lamprey. Therefore, the study on the origin and evolution of vertebrate adaptive immune system is further promoted.
【学位授予单位】：辽宁师范大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R392

【共引文献】