Yamashita T, Kasuya S, Nagano I, Ohtomo H
Department of Parasitology, Gifu University School of Medicine.
Kansenshogaku Zasshi. 1992 Sep;66(9):1262-9. doi: 10.11150/kansenshogakuzasshi1970.66.1262.
We characterized 8 monoclonal antibodies (MAbs) to Karp, Kato, and Gilliam strains of Rickettsia tsutsugamushi, and analysed 17 isolates from patients with Tsutsugamushi disease using these MAbs. These were divided into 3 strain-specific (Kp/D11, Kt/2D9, and Gi/E4) and 5 cross-reactive MAbs (Kp/1F11, Kp/1C10, Kp/C6, Kt/3B2, and Kt/3C2). All MAbs recognized characteristic protein antigens using the indirect fluorescent-antibody test (IFA) and proteinase K treatment. Analysis by polyacrylamide gel electrophoresis and immunoblotting techniques revealed that Kato-specific MAb Kt/2D9 recognized a polypeptide with a molecular mass of 54 kilodalton (kDa) of the homologous strain, and cross-reactive MAbs Kp/1F11, Kp/C6, and Kt/3B2 recognized those of 46-47 kDa, 46-47 KDa, and 60 kDa, respectively to the homologous and heterologous strains. MAbs Kp/1C10 which exhibited a high IFA titer against the Karp strain and only low titers against heterologous strains recognized only the 110 kDa polypeptide of the homologous strain. MAb Kt/3C2 which reacted with both Karp and Kato strains recognized a 54 to 56 kDa polypeptide band of the two prototype strains as well as several other polypeptides, however, each molecular mass was present in only one of two strains. Testing by the plaque reduction technique showed another characteristic of MAb Kt/3C2 to neutralize both Karp and Kato Strains. Fourteen isolated strains from patients in the south and west regions of Gifu Prefecture, the Shimokoshi stain isolated in Niigata Prefecture, and Kawasaki and Kuroki stains isolated in Miyazaki Prefecture were examined for reactivities to 8 MAbs by IFA to classify their antigenicities. No isolated strains reacted with Karp-specific Kp/D11, Kato-specific Kt/2D9, or Gilliam-specific Gi/E4.(ABSTRACT TRUNCATED AT 250 WORDS)
我们鉴定了8种针对恙虫病东方体Karp、Kato和Gilliam株的单克隆抗体(MAb),并使用这些单克隆抗体分析了17例恙虫病患者的分离株。这些单克隆抗体分为3种菌株特异性(Kp/D11、Kt/2D9和Gi/E4)和5种交叉反应性单克隆抗体(Kp/1F11、Kp/1C10、Kp/C6、Kt/3B2和Kt/3C2)。所有单克隆抗体通过间接荧光抗体试验(IFA)和蛋白酶K处理识别特征性蛋白质抗原。聚丙烯酰胺凝胶电泳和免疫印迹技术分析表明,Kato特异性单克隆抗体Kt/2D9识别同源菌株中一条分子量为54千道尔顿(kDa)的多肽,交叉反应性单克隆抗体Kp/1F11、Kp/C6和Kt/3B2分别识别同源和异源菌株中分子量为46 - 47 kDa、46 - 47 kDa和60 kDa的多肽。对Karp菌株IFA效价高而对异源菌株效价低的单克隆抗体Kp/1C10仅识别同源菌株的110 kDa多肽。与Karp和Kato菌株均反应的单克隆抗体Kt/3C2识别两种原型菌株的一条54至56 kDa多肽带以及其他几条多肽带,然而,每种分子量仅存在于两种菌株中的一种。噬斑减少技术检测显示单克隆抗体Kt/3C2的另一个特征是能中和Karp和Kato菌株。通过IFA检测岐阜县南部和西部地区患者的14株分离株、新潟县分离的下越菌株以及宫崎县分离的川崎和黑木菌株对8种单克隆抗体的反应性,以分类其抗原性。没有分离株与Karp特异性的Kp/D11、Kato特异性的Kt/2D9或Gilliam特异性的Gi/E4反应。(摘要截短于250字)