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Serologic Studies on Tyzzer's Disease

泰泽氏病的血清学研究

基本信息

批准号：
59480084
负责人：
FUJIWARA Kosaku
金额：
$ 3.71万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1984
资助国家：
日本
起止时间：
1984 至 1986
项目状态：
已结题

项目摘要

Serologic studies on Tyzzer's disease world-wide spreading in various species of animal were made and the following results were obtained:1) By complement fixation, gel diffusion, immunofluorescence as well as mouse protection tests using mouse or rat antisera and microbial antigens from infected mouse liver, close antigenic relatedness was revealed between a mouse strain from Denmark and that from Japan, between these murine strains and rabbit strains from America and West Germany as well as of a Mongolian gerbil strain from West Germany. The hamster and kitten strains from Japan, however, were antigenically far from the murine or rabbit strains. 2) Enzyme-linked immunosorbent assay (ELISA) revealed a good correlation on titers between immunofluorescence and ELISA, which was shown to be useful for studying antigenic relatedness among different bacterial strains. 3) Two monoclonal antibodies (MAb) to the RT strain of rat origin, were established, and one of the IgG2a MAb recognized an bacterial antigen having a molecular weight (M.W.) of 54,000 and combined with the bacterial flagella, while the other IgM recognized another antigen with M.W. of 73,000 and combined with the bacterial cell surface. Both MAb were protective for the challenge infection in mice in mouse hepatocyte culture. 4) Using immunoperoxidase method microbial antigen was first detected on Day 2 postinoculation (p.i.) within very small necrotized foci. When necrotic foci increased in size and number, many immunoperoxidase- positive bacteria were present within seemingly living hepatocytes at the periphery of lesions but not within the necrotized foci. Apart from large- sized necrotized foci some hepatocytes carrying abundant bacterial antigen were singly present without inducing any mesenchymal response. Electron microscopy revealed that antibody specifically combined with the flagella and bacterial surface.

通过补体固定、凝胶扩散、免疫荧光以及用小鼠或大鼠抗血清和感染小鼠肝脏的微生物抗原进行小鼠保护试验，发现来自丹麦的一株小鼠与来自日本的一株小鼠具有密切的抗原亲缘关系。这些鼠系与来自美国和西德的兔系以及来自西德的蒙古沙鼠系之间的差异。然而，来自日本的仓鼠和小猫毒株在抗原性上与小鼠或家兔毒株相差甚远。2)酶联免疫吸附试验（ELISA）显示免疫荧光与酶联免疫吸附试验的效价具有良好的相关性，可用于研究不同菌株之间的抗原相关性。3)建立了两种针对大鼠源RT菌株的单克隆抗体（MAb），其中一种IgG2a单克隆抗体识别分子量为54,000的细菌抗原并与细菌鞭毛结合，另一种IgM识别分子量为73,000的细菌抗原并与细菌细胞表面结合。在小鼠肝细胞培养中，这两种单抗对小鼠的攻击感染均有保护作用。4)免疫过氧化物酶法在接种后第2天（p.i）在非常小的坏死灶内首次检测微生物抗原。当坏死灶的大小和数量增加时，许多免疫过氧化物酶阳性细菌存在于病灶周围看似活的肝细胞内，而不在坏死灶内。除了大的坏死灶外，一些携带大量细菌抗原的肝细胞单独存在，没有引起任何间质反应。电镜观察发现，该抗体与鞭毛和细菌表面特异性结合。