Department of Basic and Industrial Microbiology, Trakya University Faculty of Science, Edirne, Turkey.
Department of Internal Medicine Rheumatology, University of Medicine and Dentistry of New Jersey, New Brunswick, USA.
Balkan Med J. 2013 Jun;30(2):215-24. doi: 10.5152/balkanmedj.2013.7059. Epub 2013 Jun 1.
The adhesion process of Borrelia burgdorferi to susceptible host cell has not yet been completely understood regarding the function of OspA, OspB and OspC proteins and a conflict exists in the infection process.
The adhesion rates of pathogenic (low BSK medium passaged or susceptible rat joint tissue co-cultivated) or non-pathogenic Borrelia burgdorferi (high BSK medium passaged) isolate (FNJ) to human umbilical vein endothelial cells (HUVEC) cultured on coverslips and the synthesis of OspA and OspC proteins were investigated to analyze the infection process of this bacterium.
In-vitro study.
Spirochetes were cultured in BSK medium or in a LEW/N rat tibiotarsal joint tissue feeder layer supported co-culture system using ESG co-culture medium and labelled with 3H-adenine for 48 hours. SDS-PAGE, Western Blotting, Immunogold A labeling as well as radiolabeling experiments were used to compare pathogenic or non pathogenic spirochetes during the adhesion process.
Tissue co-cultured B. burgdorferi adhered about ten times faster than BSK-grown spirochetes. Trypsin inhibited attachment to HUVEC and co-culture of trypsinized spirochetes with tissues reversed the inhibition. Also, the synthesis of OspC protein by spirochetes was increased in abundance after tissue co-cultures, as determined by SDS-PAGE and by electron microscopy analysis of protein A-immunogold staining by anti-OspC antibodies. OspA protein was synthesized in similar quantities in all Borrelia cultures analyzed by the same techniques.
Low BSK passaged or tissue co-cultured pathogenic Lyme disease spirochetes adhere to HUVEC faster than non-pathogenic high BSK passaged forms of this bacterium. Spirochetes synthesized OspC protein during host tissue-associated growth. However, we did not observe a reduction of OspA synthesis during host tissue co-cultivation in vitro.
尽管 OspA、OspB 和 OspC 蛋白的功能尚未完全了解，但博氏疏螺旋体对易感宿主细胞的黏附过程，以及感染过程中存在的冲突。
致病性（低 BSK 传代或易感大鼠关节组织共培养）或非致病性博氏疏螺旋体（高 BSK 传代）分离株（FNJ）对培养在盖玻片上的人脐静脉内皮细胞（HUVEC）的黏附率，以及 OspA 和 OspC 蛋白的合成，以分析该细菌的感染过程。
体外研究。
用 3H-腺嘌呤标记螺旋体，在 BSK 培养基或 LEW/N 大鼠胫跗关节组织饲养层支持的共培养系统中培养 ESG 共培养液 48 小时。用 SDS-PAGE、Western Blotting、免疫胶体金标记和放射性标记实验比较了黏附过程中的致病性或非致病性螺旋体。
组织共培养的博氏疏螺旋体的黏附速度比 BSK 培养的螺旋体快约 10 倍。胰蛋白酶抑制了对 HUVEC 的附着，而胰蛋白酶化的螺旋体与组织的共培养逆转了抑制作用。此外，通过 SDS-PAGE 和蛋白质 A-免疫胶体金染色的电子显微镜分析，发现组织共培养后，OspC 蛋白的合成量大量增加。用相同的技术分析所有博氏疏螺旋体培养物，发现 OspA 蛋白的合成量相似。
低 BSK 传代或组织共培养的致病性莱姆病螺旋体对 HUVEC 的黏附速度比该细菌的非致病性高 BSK 传代形式更快。螺旋体在宿主组织相关生长过程中合成 OspC 蛋白。然而，我们没有观察到在体外与宿主组织共培养过程中 OspA 合成的减少。
