Development of model system to analyze the antigen epitope to induce cell-mediated immunity in fish

开发模型系统来分析抗原表位以诱导鱼类细胞介导的免疫

基本信息

批准号：
12460088
负责人：
NAKANISHI Teruyuki
金额：
$ 8.26万
依托单位：
Nihon University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12460088/
关键词：
Cell-mediated immunity Cytotoxicity test MHC class I IHN virus DNA vaccination Rainbow trout Ginbuna crucian carp Mixed leukocyte culture )ニジマス DNAフィンープリント法モノクローナル抗体トランスフェクション MHC クラスI ウイルス長期培養細胞ギンブナヒラメ

项目摘要

Fish possess specific antigen recognition molecules as well as B and T lymphocytes and can evoke specific immune responses against a variety of antigens. In fish, however, information on cell-mediated immunity is limited when compared to those on humoral immunity due to the lack of in vivo and in vitro model systems. Vaccine against viral disease is limited in fish despite that viral diseases are serious problem in aquaculture. Development of such vaccines requires identification and characterization of viral antigens which induce protective immunity. It is well known that cell-mediated killing is most important defense mechanisms in the control of viral disease. The objective of this research project is to develop experimental model systems to analyze the protective antigens or epitopes involved in cell-mediated immunity for the development of fish vaccines.In collaboration with Dr. Okamoto at Tokyo University of Fisheries and Dr. Ototake at National Research Institute of Aquaculture … More we have established several assay systems for specific cell-mediated immunity, particularly on cytotoxic T-cell function employing clonal ginbuna and a syngeneic cell line. Those include the graft-versus host reaction (GVHR) and cell-mediated cytotoxicity (CMC) against allogeneic erythrocytes or cell lines. In the present research project we further pursued the studies and have established model systems for CMC against virus-infected cells and analyzed the protection mechanism using isogeneic ginbuna and cell lines together with fish viruses. We also extensively analyzed the MHC polymorphism in rainbow trout and found that trout cell line (RTG-2) shares the same MHC class I allele with homozygous clonal rainbow trout (C25 clone). We demonstrated MHC-restricted cell-mediated cytotoxicity against virus-infected cell lines using lymphocytes from C25 clonal trout sensitized by DNA immunization with IHNV-G or by infection with IHNV and RTG-2 cells as target. Experimental model systems established in the present study can be applied to investigate the protective antigens or epitopes involved in cell-mediated immunity for the development of fish vaccines as well as to investigate the recognition and killing g mechanism of specific cell-mediated immunity. Less

鱼具有特定的抗原识别分子以及B和T淋巴细胞，可以引起针对各种抗原的特定免疫复杂。然而，在FISH中，由于缺乏体内和体外模型系统，有关细胞介导的免疫学的信息与体液免疫学相比受到限制。针对病毒疾病的疫苗在鱼类目的地受到限制，即病毒疾病在水产养殖中是严重的问题。这种疫苗的开发需要鉴定和表征病毒抗原，这些病毒抗原诱导受保护的免疫组织化学物质。众所周知，细胞介导的杀戮是控制病毒疾病的最重要防御机制。该研究项目的目的是开发实验模型系统，以分析用于开发鱼类疫苗的细胞介导的免疫组织化学的受保护的抗原或表位。克隆金布纳氏菌和同步细胞系。这些包括针对同种异体红细胞或细胞系的移植物抗宿主反应（GVHR）和细胞介导的细胞毒性（CMC）。在本研究项目中，我们进一步进行了研究，并建立了针对感染病毒细胞的CMC模型系统，并使用ISEOGONIC GINBUNA和细胞系与鱼类病毒一起分析了保护机制。我们还广泛分析了彩虹鳟鱼中的MHC多态性，发现鳟鱼细胞系（RTG-2）与纯合克隆彩虹鳟鱼（C25克隆）共享了相同的MHC I类等位基因。我们使用C25克隆鳟鱼对病毒感染细胞系的MHC限制细胞介导的细胞毒性对病毒感染的细胞系进行了DNA免疫抑制敏感的淋巴细胞，并通过IHNV-G或通过IHNV和RTG-2细胞作为靶标感染。本研究中建立的实验模型系统可用于研究针对细胞介导的免疫抑制的受保护的抗原或表位，以开发鱼类疫苗，并研究特定细胞介导的免疫抑制的识别和杀死G机制。较少的