Wilske B, Fingerle V, Preac-Mursic V, Jauris-Heipke S, Hofmann A, Loy H, Pfister H W, Rössler D, Soutschek E
Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Universität München, Germany.
Med Microbiol Immunol. 1994 Feb;183(1):43-59. doi: 10.1007/BF00193630.
Immunodominant proteins are variable in molecular and antigenic structure among different genospecies of Borrelia burgdorferi sensu lato. We have recently developed an immunoblot using five recombinant antigens: the chromosomal-encoded B. burgdorferi proteins p100, the flagellin and an internal flagellin fragment thereof, and the plasmid-encoded outersurface proteins A (OspA) and C (OspC). In the present study the same antigens (derived from strain PKo, genospecies B. afzelii) were compared with the homologous recombinant proteins from strain B31 (genospecies B. burgdorferi sensu stricto) and with OspA, OspC and the internal flagellin fragment from strain PBi (genospecies B. garinii). Patients with neuroborreliosis (n = 28) and patients with acrodermatitis chronica atrophicans (n = 20) were investigated in the IgG immunoblot; the IgM immunoblot was performed only in patients with neuroborreliosis. There was a small increase in the detection rate of OspA-specific IgG or IgM antibodies using the different variants of recombinant OspA; however, OspA remained an insensitive antigen for antibody detection in Lyme borreliosis. The same was true to OspC-specific IgG antibodies. The sensitivity of OspC, which is the immunodominant antigen for IgM antibody detection, could not be increased using recombinant antigens derived from different strains. However, some sera which were negative in the recombinant immunoblot reacted with OspC in the conventional immunoblot using B. burgdorferi whole cell lysate as antigen. The most unexpected finding was the high degree of immunological heterogeneity of the internal flagellin fragments: IgG antibodies were detected in 18 of 48 patients using B31 fragments, in 25 of 48 using PKo fragments, in 23 of 48 using PBi fragments versus 33 of 48 when the three recombinant proteins were combined. PKo-derived fragments were more sensitive for antibody detection in patients with acrodermatitis chronica atrophicans, B31- and PBi-derived fragments for antibody detection in patients with neuroborreliosis. This is in agreement with the fact that isolates from patients with neuroborreliosis are predominantly belonging to the genospecies B. burgdorferi sensu stricto and B. garinii. For detection of IgM antibodies in sera from patients with neuroborreliosis, recombinant internal fragments derived from strains B31 and PBi were more sensitive than the PKo-derived fragment. The best discrimination between neuroborreliosis sera and control sera was achieved when the IgM blot was performed using recombinant internal flagellin fragments derived from strains PKo and PBi and OspC derived from B31 or PKo.
免疫显性蛋白在狭义伯氏疏螺旋体不同基因种之间的分子和抗原结构上存在差异。我们最近开发了一种免疫印迹法，使用五种重组抗原：染色体编码的伯氏疏螺旋体蛋白p100、鞭毛蛋白及其内部鞭毛蛋白片段，以及质粒编码的外表面蛋白A（OspA）和C（OspC）。在本研究中，将相同的抗原（源自PKo菌株，基因种阿氏疏螺旋体）与来自B31菌株（狭义伯氏疏螺旋体基因种）的同源重组蛋白，以及来自PBi菌株（伽氏疏螺旋体基因种）的OspA、OspC和内部鞭毛蛋白片段进行了比较。对28例神经型莱姆病患者和20例慢性萎缩性肢端皮炎患者进行了IgG免疫印迹检测；仅对神经型莱姆病患者进行了IgM免疫印迹检测。使用不同变体的重组OspA检测OspA特异性IgG或IgM抗体的检出率略有增加；然而，OspA在莱姆病螺旋体病抗体检测中仍然是一种不敏感的抗原。OspC特异性IgG抗体也是如此。OspC是IgM抗体检测的免疫显性抗原，使用来自不同菌株的重组抗原并不能提高其敏感性。然而，一些在重组免疫印迹中呈阴性的血清，在以伯氏疏螺旋体全细胞裂解物为抗原的传统免疫印迹中与OspC发生反应。最出乎意料的发现是内部鞭毛蛋白片段存在高度的免疫异质性：使用B31片段时，48例患者中有18例检测到IgG抗体；使用PKo片段时，48例中有25例；使用PBi片段时，48例中有23例；而将三种重组蛋白组合使用时，48例中有33例。源自PKo的片段对慢性萎缩性肢端皮炎患者的抗体检测更敏感，源自B31和PBi的片段对神经型莱姆病患者的抗体检测更敏感。这与神经型莱姆病患者的分离株主要属于狭义伯氏疏螺旋体和伽氏疏螺旋体基因种这一事实相符。对于检测神经型莱姆病患者血清中的IgM抗体，源自B31和PBi菌株的重组内部片段比源自PKo的片段更敏感。当使用源自PKo和PBi菌株的重组内部鞭毛蛋白片段以及源自B31或PKo的OspC进行IgM印迹检测时，神经型莱姆病血清与对照血清之间的区分效果最佳。
