Wilske B, Preac-Mursic V, Jauris S, Hofmann A, Pradel I, Soutschek E, Schwab E, Will G, Wanner G
Max von Pettenkofer Institut für Hygiene und Medizinische Mikrobiologie, Universität München, Germany.
Infect Immun. 1993 May;61(5):2182-91. doi: 10.1128/iai.61.5.2182-2191.1993.
The gene of the immunodominant major protein pC of Borrelia burgdorferi was previously cloned and sequenced (R. Fuchs, S. Jauris, F. Lottspeich, V. Preac-Mursic, B. Wilske, and E. Soutschek, Mol. Microbiol. 6:503-509, 1992). pC is abundantly expressed on the outer surface of B. burgdorferi, as demonstrated by immunoelectron microscopy with monoclonal antibody L22 1F8. Accordingly, pC is renamed OspC, by analogy to the outer surface proteins OspA and OspB. Western immunoblot analysis of 45 B. burgdorferi isolates with monoclonal antibodies revealed that OspC is immunologically heterogeneous. Partial sequence analysis of the ospC gene confirmed the protein heterogeneity at the genetic level. We found that the degree of identity between the ospC partial sequences of five strains representing different OspA serotypes was only 63.3 to 85.4%. Immunological heterogeneity was also observed among representatives of the three newly designated genospecies of B. burgdorferi sensu lato, B. burgdorferi sensu stricto, B. garinii, and group VS461. Heterogeneity was confirmed for B. garinii at the genetic level. The ospC gene was also cloned from strains that did not express OspC, and antibody-reactive OspC was expressed in Escherichia coli. In addition, OspC-expressing variants were obtained from a nonexpressing strain by plating single colonies on solid medium. These findings confirm that the ospC gene is also present in nonexpressing strains. Because OspC is an immunodominant protein for the early immune response in Lyme borreliosis and was effective as a vaccine in an animal model, the immunological and molecular polymorphisms of ospC and OspC have important implications for the development of diagnostic reagents and vaccines.
伯氏疏螺旋体免疫显性主要蛋白pC的基因先前已被克隆和测序（R. 富克斯、S. 尤里西斯、F. 洛茨皮希、V. 普雷阿克 - 穆尔西克、B. 威尔斯克和E. 绍切克，《分子微生物学》6:503 - 509，1992年）。如用单克隆抗体L22 1F8进行免疫电子显微镜观察所示，pC在伯氏疏螺旋体的外表面大量表达。因此，仿照外表面蛋白OspA和OspB，pC被重新命名为OspC。用单克隆抗体对45株伯氏疏螺旋体分离株进行的Western免疫印迹分析表明，OspC在免疫上具有异质性。ospC基因的部分序列分析在基因水平上证实了蛋白质的异质性。我们发现，代表不同OspA血清型的5个菌株的ospC部分序列之间的同一性程度仅为63.3%至85.4%。在狭义伯氏疏螺旋体的三个新指定基因种的代表菌株中也观察到了免疫异质性，即狭义伯氏疏螺旋体、伽氏疏螺旋体和VS461组。伽氏疏螺旋体在基因水平上的异质性得到了证实。ospC基因也从不表达OspC的菌株中克隆出来，并且在大肠杆菌中表达了具有抗体反应性的OspC。此外，通过将单个菌落接种在固体培养基上，从不表达的菌株中获得了表达OspC的变体。这些发现证实，ospC基因也存在于不表达的菌株中。由于OspC是莱姆病早期免疫反应中的一种免疫显性蛋白，并且在动物模型中作为疫苗有效，因此ospC和OspC的免疫和分子多态性对诊断试剂和疫苗的开发具有重要意义。
