Bunk Sebastian, Susnea Iuliana, Rupp Jan, Summersgill James T, Maass Matthias, Stegmann Werner, Schrattenholz André, Wendel Albrecht, Przybylski Michael, Hermann Corinna
Department of Biochemical Pharmacology, University of Konstanz, Konstanz, Germany.
J Immunol. 2008 Apr 15;180(8):5490-8. doi: 10.4049/jimmunol.180.8.5490.
The controversial discussion about the role of Chlamydia pneumoniae in atherosclerosis cannot be solved without a reliable diagnosis that allows discrimination between past and persistent infections. Using a proteomic approach and immunoblotting with human sera, we identified 31 major C. pneumoniae Ags originating from 27 different C. pneumoniae proteins. More than half of the proteins represent Chlamydia Ags not described previously. Using a comparative analysis of spot reactivity Pmp6, OMP2, GroEL, DnaK, RpoA, EF-Tu, as well as CpB0704 and CpB0837, were found to be immunodominant. The comparison of Ab-response patterns of sera from subjects with and without evidence for persisting C. pneumoniae, determined by multiple PCR analysis of PBMC and vasculatory samples, resulted in differential reactivity for 12 proteins, which is not reflected by reactivity of the sera in the microimmunofluorescence test, the current gold standard for serodiagnosis. Although reactivity of sera from PCR-positive donors was increased toward RpoA, MOMP, YscC, Pmp10, PorB, Pmp21, GroEL, and Cpaf, the reactivity toward YscL, Rho, LCrE, and CpB0837 was decreased, reflecting the altered protein expression of persisting C. pneumoniae in vitro. Our data provide the first evidence of a unique Ab-response pattern associated with persistent C. pneumoniae infections, which is a prerequisite for the serological determination of persistently infected patients.
关于肺炎衣原体在动脉粥样硬化中作用的争议性讨论,如果没有可靠的诊断方法来区分既往感染和持续性感染,就无法得到解决。我们采用蛋白质组学方法并结合人血清免疫印迹,鉴定出了源自27种不同肺炎衣原体蛋白的31种主要肺炎衣原体抗原。超过半数的蛋白代表此前未描述过的衣原体抗原。通过斑点反应性的比较分析,发现Pmp6、OMP2、GroEL、DnaK、RpoA、EF-Tu以及CpB0704和CpB0837具有免疫优势。对通过PBMC和血管样本多重PCR分析确定的有或无持续性肺炎衣原体感染证据的受试者血清抗体反应模式进行比较,结果显示12种蛋白具有不同的反应性,而这在血清学诊断的现行金标准——微量免疫荧光试验中并未体现。尽管PCR阳性供体的血清对RpoA、主要外膜蛋白(MOMP)、YscC、Pmp10、PorB、Pmp21、GroEL和衣原体蛋白酶样活性因子(Cpaf)的反应性增强,但对YscL、Rho、LCrE和CpB0837的反应性降低,这反映了体外持续性肺炎衣原体蛋白质表达的改变。我们的数据首次证明了与持续性肺炎衣原体感染相关的独特抗体反应模式,这是血清学检测持续性感染患者的先决条件。