Jones S A, Marchitto K S, Miller J N, Norgard M V
J Exp Med. 1984 Nov 1;160(5):1404-20. doi: 10.1084/jem.160.5.1404.
Radioimmunoprecipitation (RIP) analyses performed on 125I-surface-labeled Treponema pallidum cells using various immune sera revealed the presence of six major surface antigens (immunogens) with apparent molecular weights of 47 K, 36 K, 34 K, 32 K, 29 K, and 13 K. Among these, the 47 K surface antigen was most abundant. Radioimmunoprecipitation assays using 125I-labeled T. phagedenis biotype Reiter or immunoblot analyses using the same strain, failed to reveal the presence of the 47 K mol wt antigen in the representative nonpathogenic treponeme. Preabsorption of anti-T. pallidum immune rabbit serum (IRS) with the Reiter organism did not remove anti-T. pallidum antibodies from immune serum that reacted with the 47 K mol wt immunogen or other immunogens of T. pallidum present in the characteristic antigenic profile. Monoclonal antibodies (mAb) directed specifically against the 47 K mol wt immunogen of T. pallidum also failed to react with an analogous 47 K mol wt component in Treponema phagedenis biotype Reiter, further suggesting the unique presence of this antigen in pathogenic treponemes. The presence of the 47 K mol wt surface immunogen in pathogenic treponemes other than T. pallidum subspecies pallidum was also observed (43). Anti-47 K immunogen mAb was nonreactive against rabbit IgG or IgM. mAb directed specifically against the 47 K mol wt immunogen of T. pallidum was examined for strategic functional activities. It was found to be reactive in the microhemagglutination assay for T. pallidum antibodies, the T. pallidum immobilization test, and was found to be capable of significant blockage of attachment of virulent T. pallidum to host cells in tissue culture. Additional significant biological activity for the anti-47 K mol wt immunogen mAb was revealed through results of the in vitro-in vivo neutralization test of Bishop and Miller, in which a 99% or 100% neutralizing activity was demonstrated. The combined data of this study suggest that the 47 K mol wt immunogen of T. pallidum represents an abundant, immunodominant, surface-exposed immunogen possessing potential biological importance in the pathogenesis and immunology of T. pallidum infection. These studies serve to establish the first functionally defined immunogen for T. pallidum, which may represent the major immunogen of the organism.
使用各种免疫血清对125I表面标记的梅毒螺旋体细胞进行放射免疫沉淀(RIP)分析,结果显示存在六种主要表面抗原(免疫原),其表观分子量分别为47K、36K、34K、32K、29K和13K。其中,47K表面抗原最为丰富。使用125I标记的噬菌密螺旋体生物变种赖特氏株进行放射免疫沉淀测定,或使用同一菌株进行免疫印迹分析,均未在代表性非致病性密螺旋体中发现47K分子量抗原的存在。用赖特氏株对抗梅毒螺旋体免疫兔血清(IRS)进行预吸收,并未从与47K分子量免疫原或梅毒螺旋体特征性抗原谱中存在的其他免疫原发生反应的免疫血清中去除抗梅毒螺旋体抗体。针对梅毒螺旋体47K分子量免疫原的单克隆抗体(mAb)也未与噬菌密螺旋体生物变种赖特氏株中类似的47K分子量成分发生反应,这进一步表明该抗原在致病性密螺旋体中独特存在。在梅毒亚种苍白亚种以外的致病性密螺旋体中也观察到了47K分子量表面免疫原的存在(43)。抗47K免疫原mAb与兔IgG或IgM无反应。对针对梅毒螺旋体47K分子量免疫原的mAb进行了战略功能活性检测。发现它在梅毒螺旋体抗体的微量血凝试验、梅毒螺旋体固定试验中具有反应性,并且发现在组织培养中能够显著阻断强毒梅毒螺旋体与宿主细胞的附着。通过毕晓普和米勒的体外-体内中和试验结果,揭示了抗47K分子量免疫原mAb的其他显著生物学活性,其中显示出99%或100%的中和活性。本研究的综合数据表明,梅毒螺旋体47K分子量免疫原是一种丰富的、免疫显性的、表面暴露的免疫原,在梅毒螺旋体感染的发病机制和免疫学中具有潜在的生物学重要性。这些研究有助于确立梅毒螺旋体的首个功能明确的免疫原,它可能代表该生物体的主要免疫原。