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首页> 外文期刊>Infection and immunity >A Monoclonal Antibody Generated by Antigen Inoculation via Tick Bite Is Reactive to the Borrelia burgdorferi Rev Protein, a Member of the 2.9 Gene Family Locus
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A Monoclonal Antibody Generated by Antigen Inoculation via Tick Bite Is Reactive to the Borrelia burgdorferi Rev Protein, a Member of the 2.9 Gene Family Locus

机译：通过via虫抗原接种产生的单克隆抗体可与博雷利亚氏疏螺旋体Rev蛋白（2.9基因家族基因的成员）反应

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Murine monoclonal antibodies directed against proteins ofBorrelia burgdorferi B31 (low passage) were generated by the administration of antigen via the bite of borrelia-infected ticks. This strategy was employed as a mechanism to create antibodies against antigens presented by the natural route of tick transmission versus those presented by inoculation with cultured borreliae. One of the resultant antibodies reacted with a 17-kDa antigen from culturedB. burgdorferi, as seen by immunoblot analysis. This antibody was used to screen a B. burgdorferi genomic DNA lambda vector expression library, and an immunoreactive clone was isolated. DNA sequence analysis of this clone, containing a 2.7-kb insert, revealed several open reading frames. These open reading frames were found to be homologs of genes discovered as a multicopy gene family in the 297 strain of B. burgdorferi by Porcella et al. (S. F. Porcella, T. G. Popova, D. R. Akins, M. Li, J. D. Radolf, and M. V. Norgard, J. Bacteriol. 178:3293–3307, 1996). By selectively subcloning genes found in this insert into an Escherichia coli plasmid expression vector, the observation was made that the rev gene product was the protein reactive with the 17-kDa-specific monoclonal antibody. Therev gene product was found to be expressed in low-passage, but not in high-passage, B. burgdorferi B31. Correspondingly, the rev gene was not present in strain B31 genomic DNA from cultures that had been passaged >50 times. Serum samples from Lyme disease patients demonstrated an antibody response against the Rev protein. The generation of an anti-Rev response in Lyme disease patients, and in mice by tick bite inoculation, provides evidence that the Rev protein is expressed and immunogenic during the course of natural transmission and infection.
机译：通过博瑞氏菌感染的tick叮咬给予抗原，产生了针对博氏疏螺旋体B31蛋白的鼠单克隆抗体（低传代）。该策略被用作产生针对由create传播的天然途径呈现的抗原的抗体的机制，该抗与通过接种培养的疏螺旋体呈现的抗原的抗体。所得抗体之一与来自培养的B的17-kDa抗原反应。通过免疫印迹分析可以看到burgdorferi 。该抗体用于筛选 B。 burgdorferi基因组DNA lambda载体表达文库，并分离出免疫反应性克隆。该克隆的DNA序列分析（包含一个2.7-kb的插入片段）揭示了几个开放阅读框。发现这些开放阅读框是在297株 B中作为多拷贝基因家族发现的基因的同源物。 Porcella等人的burgdorferi 。 （S. F. Porcella，T. G. Popova，D. R. Akins，M. Li，J. D. Radolf和M. V. Norgard，J. Bacteriol。178：3293-3307，1996）。通过选择性地将该插入物中发现的基因亚克隆到大肠杆菌质粒表达载体中，观察到 rev 基因产物是与17-kDa特异性反应的蛋白。单克隆抗体。发现 rev 基因产物以低传代 B表达，但不以高传代 B表达。 burgdorferi B31。相应地， rev 基因在传代> 50次的培养物中的菌株B31基因组DNA中不存在。来自莱姆病患者的血清样品显示出针对Rev蛋白的抗体反应。在莱姆病患者中以及通过tick叮咬接种在小鼠中产生抗Rev反应，提供了证据，表明Rev蛋白在自然传播和感染过程中表达并具有免疫原性。

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