Wilske B, Jauris-Heipke S, Lobentanzer R, Pradel I, Preac-Mursic V, Rössler D, Soutschek E, Johnson R C
Max von Pettenkofer Institut für Hygiene und Medizinische Mikrobiologie, Universität München, Germany.
J Clin Microbiol. 1995 Jan;33(1):103-9. doi: 10.1128/jcm.33.1.103-109.1995.
Molecular analyses of the genes encoding OspC, a major immunodominant protein of Borrelia burgdorferi sensu lato, revealed a considerable degree of heterogeneity. In the present study, we investigated whether a similar heterogeneity of the OspC phenotype can be shown by analysis with monoclonal antibodies (MAbs). Thirteen OspC-specific MAbs (L22 MAbs) were produced by immunizing mice with either different combinations of whole-cell antigens or recombinantly expressed OspCs cloned from strains belonging to different Borrelia spp. Ten of them differed in their reactivities with various strains. Western blot (immunoblot) analyses of 38 B. burgdorferi sensu lato strains resulted in 13 different reactivity patterns. These 13 different patterns were observed among only six different OspA serotypes, indicating that OspC is more heterogeneous than OspA. Patterns 1 to 4 were present only in B. burgdorferi sensu stricto, patterns 5 to 7 were present only in Borrelia afzelii, and patterns 9 to 13 were present only in Borrelia garinii. Pattern 8 was observed among B. afzelii and B. garinii strains but not among B. burgdorferi sensu stricto strains. One L22 MAb (2B8) recognized a common OspC-specific epitope of all 38 B. burgdorferi sensu lato strains analyzed, and another one (22C11) recognized a common epitope of OspC from both B. afzelii and B. garinii and was not reactive with OspC from B. burgdorferi sensu stricto. Western blot and sequence analysis of truncated OspCs located the 22C11 epitope as well as a species-specific sequence motif between amino acids 20 and 35. Other broadly reactive L22 MAbs were 10D3, 1F8, and 7G5. Some L22 MAbs (1C3, 1C3, 12E5, 1B11, 1F10, and 6C8) bound to epitopes present only in a few strains. Relapsing fever borreliae (Borrelia hermsii, Borrelia turicatae, and Borrelia duttoni) were nonreactive, with the following exception: three L22 MAbs (2B8, 6C4, and 10C5) recognized an abundantly expressed 20-kDa-range protein of B. turicatae. Because OspC is an immunodominant protein during the early immune response in Lyme borreliosis and has been shown to be effective as a vaccine in an animal model, our findings have important implications for the development of diagnostic reagents as well as vaccine research.
伯氏疏螺旋体狭义种主要免疫显性蛋白OspC编码基因的分子分析显示出相当程度的异质性。在本研究中，我们调查了通过单克隆抗体（MAb）分析是否能显示出OspC表型的类似异质性。通过用全细胞抗原的不同组合或从属于不同疏螺旋体物种的菌株克隆的重组表达OspC免疫小鼠，产生了13种OspC特异性MAb（L22 MAb）。其中10种在与各种菌株的反应性上有所不同。对38株伯氏疏螺旋体狭义种菌株进行的蛋白质免疫印迹（免疫印迹）分析产生了13种不同的反应模式。仅在6种不同的OspA血清型中观察到这13种不同模式，表明OspC比OspA更具异质性。模式1至4仅存在于伯氏疏螺旋体狭义种中，模式5至7仅存在于阿氏疏螺旋体中，模式9至13仅存在于伽氏疏螺旋体中。模式8在阿氏疏螺旋体和伽氏疏螺旋体菌株中观察到，但在伯氏疏螺旋体狭义种菌株中未观察到。一种L22 MAb（2B8）识别了所分析的所有38株伯氏疏螺旋体狭义种菌株共有的OspC特异性表位，另一种（22C11）识别了阿氏疏螺旋体和伽氏疏螺旋体OspC的共有表位，与伯氏疏螺旋体狭义种的OspC无反应。对截短的OspC进行蛋白质免疫印迹和序列分析，将22C11表位以及一个物种特异性序列基序定位在氨基酸20至35之间。其他具有广泛反应性的L22 MAb是10D3、1F8和7G5。一些L22 MAb（1C3、1C3、12E5、1B11、1F10和6C8）与仅在少数菌株中存在的表位结合。回归热疏螺旋体（赫氏疏螺旋体、杜氏疏螺旋体和图氏疏螺旋体）无反应，但有以下例外：三种L22 MAb（2B8、6C4和10C5）识别图氏疏螺旋体一种大量表达的20 kDa范围的蛋白。由于OspC是莱姆病早期免疫反应中的一种免疫显性蛋白，并且已在动物模型中显示作为疫苗有效，我们的发现对诊断试剂的开发以及疫苗研究具有重要意义。
