iSolatins of recent prevalent Rickettsia tsutsugamushi, and analysis of their protein constitution and antigenic components.

近期流行的恙虫病立克次体的iSolatins及其蛋白质组成和抗原成分分析。

• 批准号: 60570201
• 负责人: TAMURA Akira
• 金额: $ 1.15万
• 依托单位: Niigata College of Pharmacy
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-60570201/
• 关键词: Rickettsia tsutsugamushi; Protein constitution; Antigenic component; Teckettsia of new serotype; Monoclonal antibody; Surface structure; Vertical transmission; 抗生物質感受性; R.tsutsugamushi; 株特異的抗原; 蛋白の精製; 病原体の分離; 細菌表層構造

The results obtained were summatized as follows. (1) Protein constitution of Rickettsia tsutsugamushi in polyacrylamide gel electrophoresis showed some differences among the strains, but the over-all patterns resembled each other. proteins located on the rickettsial surface were identified by^<125> I labeling and others, and some of them, including 54-56K (kirodaltons) protein, showed heatmodifiability. (2) In immunoblotting analysis with guinea pig hyperimmune sera against the prototype strains of Gilliam, Karp, and Kato, 54-56K protein showed strain-specificity, and 45K and 70K proteins appeared grouyp-specificity. Monoclonal andibodies specific to each prototype strain reacted only with the 54-56K protein of homologous strain, indicating also the strain-specific antigenicity of 54-56K proteins. (3) Strains isolated recently form patients and natural chigger mites werte classifed, from the reacitivities with strain-specifif monoclonal andibodies, into Gilliam-type, Karp-type and non- … More reactive type (non-G-KP-KT Type). This indicated the existence of ewn serotype different from thr prototype strains in Japan. (4) All non-G-KP-KT type strains showed low virulence to mice. (5) Analysis of reacitivity of patrient sara with rickettsial antigens in immunoblotting tests showed variety in each serum, although many sera contained antibodies reactive with 54-56K proteins. (6) In purified rickettsial preparation, muramic acid, glucosamine, heptose and 3-deoxy-D-mannooctulosonic acid, which are constitutions of bacterial peptidoglycan (PG) ot lipopolysacchatide (LPS), were not detectred by chemical analysis, indicating that R. tsutsugamushi is lacking PG and LPS. (7) 5456K proteins were purified from the prototype strains, and the amino acid compositions and the sequences at N-terminals were compared each other. (8) Electron microscopic obserbvations of embryos of Leptotrombidium pallidum naturally infected with R. tsutsugamushi indicated that the recikettsiae were surely transmitted vertically from an adult mite to larvae through embryo, and that rickettsiae are multiplying in the embryo during the developemnt. (9) Differnce of antibiotic sensitivity berween prototype and newly isolated strain were not detected. Less

所得结果总结如下。(1)恙虫病立克次体聚丙烯酰胺凝胶电泳的蛋白质组成存在一定差异，但总的电泳图谱相似。通过~ 1H-I标记等方法鉴定了立克次体表面的蛋白质<125>，其中包括54- 56 K（kirodaltons）蛋白，它们具有热变性。(2)在对Gilliam、Karp和Kato原型菌株的豚鼠超免疫血清的免疫印迹分析中，54- 56 K蛋白表现出菌株特异性，45 K和70 K蛋白表现出群体特异性。对各原型菌株特异的单克隆抗体仅与同源菌株的54- 56 K蛋白反应，这也表明54- 56 K蛋白具有菌株特异性抗原性。(3)从病人和自然恙螨中分离到的恙螨菌株，根据其与菌株特异性单克隆抗体的反应性，可分为Gilbert型、Karp型和非Gilbert型。 ...更多信息 反应型（非G-KP-KT型）。这表明存在不同于日本原型株的母羊血清型。(4)所有非G-KP-KT型菌株对小鼠的毒力均较低。(5)在免疫印迹试验中，对阳性sara与立克次体抗原的反应性分析显示，尽管许多血清含有与54- 56 K蛋白反应的抗体，但每种血清的反应性不同。(6)在纯化的立克次体制剂中，化学分析均未检测到构成细菌肽聚糖（PG）或脂多糖（LPS）的胞壁酸、氨基葡萄糖、庚糖和3-脱氧-D-甘露辛酮糖酸，表明立克次体是一种致病性立克次体。恙虫病缺乏PG和LPS。(7)从原型菌株中纯化了5456 K蛋白，并比较了其氨基酸组成和N端序列。(8)苍白纤恙螨自然感染R.恙虫病立克次体的研究表明，立克次体确实是由成螨经胚胎垂直传播给幼虫的，立克次体在发育过程中是在胚胎中繁殖的。(9)原型菌株与新分离菌株对抗生素的敏感性无明显差异。少

项目成果

Yamamoto,S.et al.: Microbiol.Immunol.30. 611-620 (1986)

Yamamoto，S.et al.：Microbiol.Immunol.30。

大橋典男,多村憲: 日本細菌学雑誌. 42. 394 (1987)

Norio Ohashi，Ken Tamura：日本细菌学杂志 42. 394 (1987)。

Amano,Ken-ichi: Infection and Immunity. 55. 2290-2292 (1987)

天野健一：感染与免疫。

Tamura, Akira: "Analysis of polypeptide composition and antigenic components of Rickettsia tsutsugamushi by polyacrylamide gel electrophoresis and immunoblotting" Infection and Immunity. 48. 671-675 (1985)

Tamura, Akira：“通过聚丙烯酰胺凝胶电泳和免疫印迹分析恙虫病立克次体的多肽组成和抗原成分”感染与免疫。

宮村定男: 感染症学雑誌. 59. 486-488 (1985)

宫村贞夫：传染病杂志 59. 486-488 (1985)