Crother Timothy R, Champion Cheryl I, Wu Xiao-Yang, Blanco David R, Miller James N, Lovett Michael A
Department of Medicine, University of California, Los Angeles, 90095, USA.
Infect Immun. 2003 Jun;71(6):3419-28. doi: 10.1128/IAI.71.6.3419-3428.2003.
The general concept that during infection of mice the Borrelia burgdorferi surface protein composition differs profoundly from that of tick-borne or in vitro-cultivated spirochetes is well established. Specific knowledge concerning the differences is limited because the small numbers of spirochetes present in tissue have not been amenable to direct compositional analysis. In this report we describe novel means for studying the antigenic composition of host-adapted Borrelia (HAB). The detergent Triton X-114 was used to extract the detergent-phase HAB proteins from mouse ears, ankles, knees, and hearts. Immunoblot analysis revealed a profile distinct from that of in vitro-cultivated Borrelia (IVCB). OspA and OspB were not found in the tissues of SCID mice 17 days after infection. The amounts of antigenic variation protein VlsE and the relative amounts of its transcripts were markedly increased in ear, ankle, and knee tissues but not in heart tissue. VlsE existed as isoforms having both different unit sizes and discrete lower molecular masses. The hydrophobic smaller forms of VlsE were also found in IVCB. The amounts of the surface protein (OspC) and the decorin binding protein (DbpA) were increased in ear, ankle, knee, and heart tissues, as were the relative amounts of their transcripts. Along with these findings regarding VlsE, OspC, and DbpA, two-dimensional immunoblot analysis with immune sera also revealed additional details of the antigenic composition of HAB extracted from ear, heart, and joint tissues. A variety of novel antigens, including antigens with molecular masses of 65 and 30 kDa, were found to be upregulated in mouse tissues. Extraction of hydrophobic B. burgdorferi antigens from tissue provides a powerful tool for determining the antigenic composition of HAB.
在小鼠感染过程中，伯氏疏螺旋体的表面蛋白组成与蜱传播或体外培养的螺旋体有很大不同，这一普遍概念已得到充分证实。关于这些差异的具体知识有限，因为组织中存在的少量螺旋体难以进行直接的成分分析。在本报告中，我们描述了研究宿主适应性伯氏疏螺旋体（HAB）抗原组成的新方法。使用去污剂Triton X - 114从小鼠耳朵、脚踝、膝盖和心脏中提取去污剂相HAB蛋白。免疫印迹分析显示其图谱与体外培养的伯氏疏螺旋体（IVCB）不同。感染17天后，在SCID小鼠的组织中未发现OspA和OspB。抗原变异蛋白VlsE的量及其转录本的相对量在耳朵、脚踝和膝盖组织中显著增加，但在心脏组织中未增加。VlsE以具有不同单位大小和离散较低分子量的异构体形式存在。在IVCB中也发现了VlsE的疏水性较小形式。表面蛋白（OspC）和饰胶蛋白结合蛋白（DbpA）的量在耳朵、脚踝、膝盖和心脏组织中增加，其转录本的相对量也增加。除了这些关于VlsE、OspC和DbpA的发现外，用免疫血清进行的二维免疫印迹分析还揭示了从耳朵、心脏和关节组织中提取的HAB抗原组成的其他细节。发现多种新抗原，包括分子量为65和30 kDa的抗原，在小鼠组织中上调。从组织中提取疏水性伯氏疏螺旋体抗原为确定HAB的抗原组成提供了一个有力工具。
