Characterization of the outer membrane Proteome of Leptospira interrogans expressed during acute lethal infection
Characterization of the outer membrane Proteome of Leptospira interrogans expressed during acute lethal infection
复制标题
DOI：
10.1128/iai.00741-06
复制
发表时间：
2007-02-01
影响因子：
3.1
通讯作者：
Lovett, Michael A.
中科院分区：
文献类型：
--
作者：
Nally, Jarlath E.;Whitelegge, Julian P.;Lovett, Michael A.
致病性钩端螺旋体在疾病传播和感染过程中适应多种环境条件。虽然迄今为止，体外培养的钩端螺旋体的蛋白质组已在多项研究中得到表征，但对于钩端螺旋体在疾病过程中表达的蛋白质组知之甚少。从患有严重肺部钩端螺旋体病的患者身上分离得到的钩端螺旋体分离物会引起豚鼠的急性致死感染和大鼠的慢性无症状感染。最近的研究表明，从急性感染的豚鼠组织中回收的钩端螺旋体的蛋白质和脂多糖成分不同于慢性感染的大鼠组织和体外培养的钩端螺旋体（J. E. Nally、E. Chow、M. C. Fishbein、D. R. Blanco 和 M. A. Lovett，Infect. Immun. 73:3251-3260， 2005）。在当前的研究中，在用 Triton X-114 富集疏水性膜蛋白后，相对于体外培养的钩端螺旋体 (IVCL) 的蛋白质组，对疾病过程中表达的钩端螺旋体的蛋白质组进行了表征。通过二维凝胶电泳分离蛋白质样品，并通过使用单特异性抗血清和恢复期大鼠血清的免疫印迹以及质谱法来鉴定感染期间表达的抗原。结果表明，在豚鼠急性感染期间，外膜蛋白 Loa22 的表达相对于其他外膜蛋白显着增加，而其他外膜蛋白的表达相对于 IVCL 中的表达通常减少。在豚鼠急性感染期间，钩端螺旋体也表达大量 LipL32。
Pathogenic Leptospira species adapt to a wide range of environmental conditions during disease transmission and infection. While the proteome of in vitro cultivated Leptospira has been characterized in several studies to date, relatively little is known of the proteome as expressed by Leptospira during disease processes. Isolates of Leptospira obtained from patients suffering the severe pulmonary form of leptospirosis cause acute lethal infection in guinea pigs and chronic asymptomatic infection in rats. Recent studies have demonstrated that protein and lipopolysaccharide constituents of Leptospira recovered from acutely infected guinea pig tissue differ from that of Leptospira in chronically infected rat tissue and in vitro cultivated Leptospira (J. E. Nally, E. Chow, M. C. Fishbein, D. R. Blanco, and M. A. Lovett, Infect. Immun. 73:3251-3260, 2005). In the current study, the proteome of Leptospira expressed during disease processes was characterized relative to that of in vitro cultivated Leptospira (IVCL) after enrichment for hydrophobic membrane proteins with Triton X-114. Protein samples were separated by two-dimensional gel electrophoresis, and antigens expressed during infection were identified by immunoblotting with monospecific antiserum and convalescent rat serum in addition to mass spectrometry. Results suggest a significant increase in the expression of the outer membrane protein Loa22 during acute infection of guinea pigs relative to other outer membrane proteins, whose expression is generally diminished relative to expression in IVCL. Significant amounts of LipL32 are also expressed by Leptospira during acute infection of guinea pigs.