Temporal analysis of the antigenic composition of Borrelia burgdorferi during infection in rabbit skin
Temporal analysis of the antigenic composition of Borrelia burgdorferi during infection in rabbit skin
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DOI：
10.1128/iai.72.9.5063-5072.2004
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发表时间：
2004-09-01
影响因子：
3.1
通讯作者：
Lovett, MA
中科院分区：
文献类型：
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作者：
Crother, TR;Champion, CI;Lovett, MA
在感染的前3周内，通过实时PCR评估兔皮肤中宿主适应的伯氏疏螺旋体（HAB）生物体的数量。在第11天发现最大数量，而到第21天螺旋体数量减少了30倍以上。通过使用称为疏水抗原组织Triton提取的程序来确定皮肤活检样品中的HAB的抗原组成。来自兔的免疫血清、来自慢性感染小鼠的血清和抗原性变异蛋白质的单特异性抗血清，VISE，用于探测代表每个时间点的平行二维免疫印迹。使用特异性抗血清或通过将蛋白质斑点与来自体外培养的疏螺旋体的质谱鉴定的蛋白质斑点匹配来鉴定单个蛋白质。在21天的时间内，各种已知的和以前未识别的HAB抗原的相对表达有显着的变化。OspC和外膜蛋白OspA和OspB在最早的时间点，即第5天，当抗原性变异蛋白VISE几乎检测不到时是突出的。第5天后未检测到OspA和OspB。第9天后未检测到OspC。从第7天到第21天，VISE是最突出的抗原。从第7天到第21天还检测到BmpA、ErpN、ErpP、LA 7、OppA-2、DbpA和未鉴定的15-kDa蛋白。使用单特异性抗VISE的免疫印迹分析揭示了在第9、11和14天HAB中存在显著的不同的VISE较低形式;然而，在第21天不再检测到这些较低形式。可见较低形式的这种显著减少表明螺旋体从皮肤上清除。
The numbers of host-adapted Borrelia burgdorferi (HAB) organisms in rabbit skin were assessed by real-time PCR over the first 3 weeks of infection. Maximal numbers were found at day 11, while spirochete numbers decreased by more than 30-fold by day 21. The antigenic composition of HAB in skin biopsy samples was determined by use of a procedure termed hydrophobic antigen tissue Triton extraction. Immune sera from rabbits, sera from chronically infected mice, and monospecific antiserum to the antigenic variation protein, VIsE, were used to probe parallel two-dimensional immunoblots representing each time point. Individual proteins were identified using either specific antisera or by matching protein spots to mass spectrometry-identified protein spots from in vitro-cultivated Borrelia. There were significant changes in the relative expression of a variety of known and previously unrecognized HAB antigens during the 21-day period. OspC and the outer membrane proteins OspA and OspB were prominent at the earliest time point, day 5, when the antigenic variation protein VIsE was barely detected. OspA and OspB were not detected after day 5. OspC was not detected after day 9. VIsE was the most prominent antigen from day 7 through day 21. BmpA, ErpN, ErpP, LA7, OppA-2, DbpA, and an unidentified 15-kDa protein were also detected from day 7 through day 21. Immunoblot analysis using monospecific anti-VIsE revealed the presence of prominent distinct VIsE lower forms in HAB at days 9, 11, and 14; however, these lower forms were no longer detected at day 21. This marked diminution in VIsE lower forms paralleled the clearance of the spirochete from skin.