Rössler D, Eiffert H, Jauris-Heipke S, Lehnert G, Preac-Mursic V, Teepe J, Schlott T, Soutschek E, Wilske B
Max von Pettenkofer Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians Universität München, Germany.
Med Microbiol Immunol. 1995 May;184(1):23-32. doi: 10.1007/BF00216786.
The complete coding regions of the chromosomally encoded p83/100 protein of four Borrelia garinii strains and one Borrelia burgdorferi sensu stricto strain have been amplified by the polymerase chain reaction (PCR), cloned and sequenced. From alignment studies with the deduced amino acid sequences presented here, and five other published p83/100 sequences, the most heterologous region of the p83/100 molecule was identified to be located between amino acid position 390-540. To study the structure of this heterogeneous region, and internal fragment of the p83/100 genes from 11 additional B. burgdorferi sensu lato strains was amplified by PCR. The PCR products were analyzed by DNA sequencing and restriction enzyme analysis. These internal p83/100 fragments varied in size and sequence. Cluster analysis of internal p83/100 fragments, as well as restriction enzyme analysis, revealed three major groups in accordance with grouping into the three species causing Lyme disease. Strains within the same species (six B. burgdorferi sensu stricto and six B. afzelii strains) showed similar p83/100 partial structures. Nevertheless, nine B. garinii strains showed more sequence variations and could be further divided into two major subgroups. One group is represented by OspA serotype 4 strains, the other more heterogeneous group is represented by OspA serotypes 3, 5, 6 and 7 strains. Phenotypic analysis with four p83/100-specific monoclonal antibodies revealed four distinct reactivity patterns. Antibody L100 1B4 recognized a common epitope of B. burgdorferi sensu stricto and B. afzelii. Antibodies L100 17D3 and L100 18B4 were reactive with an epitope shared by strains of all three species. The broadest reactivity was shown by L100 18B4 which, in contrast to L100 17D3, additionally recognized the relapsing fever borreliae B. turicatae and B. hermsii. L100 8B8 detected a subgroup of the B. burgdorferi sensu stricto strains. Since comparison of the p83/100 molecule with sequences from protein databases showed similarities with characteristics of eukaryotic cell structures, the p83/100 might mimic these structures and may, therefore, be involved in the immune escape mechanism of the pathogenic agent of Lyme disease.
利用聚合酶链反应（PCR）扩增了4株莱姆病疏螺旋体菌株和1株狭义伯氏疏螺旋体菌株的染色体编码p83/100蛋白的完整编码区，并进行了克隆和测序。通过与本文给出的推导氨基酸序列以及其他5个已发表的p83/100序列进行比对研究，确定p83/100分子中最具异源性的区域位于氨基酸位置390 - 540之间。为研究该异质区域的结构，通过PCR扩增了另外11株狭义伯氏疏螺旋体菌株p83/100基因的内部片段。对PCR产物进行了DNA测序和限制性内切酶分析。这些p83/100内部片段在大小和序列上存在差异。对p83/100内部片段的聚类分析以及限制性内切酶分析显示，根据导致莱姆病的三种物种的分组可分为三大组。同一物种内的菌株（6株狭义伯氏疏螺旋体菌株和6株阿氏疏螺旋体菌株）显示出相似的p83/100部分结构。然而，9株莱姆病疏螺旋体菌株表现出更多的序列变异，可进一步分为两个主要亚组。一组以OspA血清型4菌株为代表，另一组更具异质性的组以OspA血清型3、5、6和7菌株为代表。用4种p83/100特异性单克隆抗体进行的表型分析揭示了4种不同的反应模式。抗体L100 1B4识别狭义伯氏疏螺旋体和阿氏疏螺旋体的一个共同表位。抗体L100 17D3和L100 18B4与所有三种物种的菌株共有的一个表位发生反应。L100 18B4显示出最广泛的反应性，与L100 17D3相比，它还识别回归热疏螺旋体土拉热疏螺旋体和赫氏疏螺旋体。L100 8B8检测到狭义伯氏疏螺旋体菌株的一个亚组。由于将p83/100分子与蛋白质数据库中的序列进行比较显示与真核细胞结构特征相似，p83/100可能模拟这些结构，因此可能参与莱姆病病原体的免疫逃逸机制。
