Yulanda M Williamson, Hercules Moura, Jennifer Whitmon, Adrian R Woolfitt, David M Schieltz, Jon C Rees, Stephanie Guo, Heather Kirkham, Daniel Bouck, Edwin W Ades, Maria Lucia Tondella, George M Carlone, Jacquelyn S Sampson, John R Barr
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引用次数: 11
摘要
百日咳博德泰拉(Bp)是百日咳(百日咳)的病原体,是一种高度传染性的感染。虽然百日咳可以通过疫苗预防,但近年来,尽管疫苗覆盖率很高,但发病率有所增加。病例增加的可能原因包括:Bp毒株适应、疫苗免疫力下降、监测加强和临床诊断改进。2010年,美国加利福尼亚州爆发百日咳;儿童和青春期前受影响最大,但疾病负担主要落在婴儿身上。为了鉴定与此次百日咳爆发相关的蛋白质生物标志物,我们利用无凝胶蛋白质组学质谱(MS)对6株Bp分离株和百日咳无细胞疫苗株Bp Tohama I (T)进行了全细胞蛋白质鉴定。MS/MS色氨酸检测和蛋白数据库检索结合western blot分析显示,本研究中3株Bp分离株的perpern(百日咳无细胞疫苗的一个亚基)的检出率显著降低。此外,利用抗bp T兔多克隆抗血清和全细胞蛋白,采用抗体亲和捕获技术鉴定推定的免疫原。蛋白质组分析可以揭示发病机制,并可能为减少感染传播策略和改善临床诊断奠定基础。
A Proteomic Characterization of Bordetella pertussis Clinical Isolates Associated with a California State Pertussis Outbreak.
Bordetella pertussis (Bp) is the etiologic agent of pertussis (whooping cough), a highly communicable infection. Although pertussis is vaccine preventable, in recent years there has been increased incidence, despite high vaccine coverage. Possible reasons for the rise in cases include the following: Bp strain adaptation, waning vaccine immunity, increased surveillance, and improved clinical diagnostics. A pertussis outbreak impacted California (USA) in 2010; children and preadolescents were the most affected but the burden of disease fell mainly on infants. To identify protein biomarkers associated with this pertussis outbreak, we report a whole cellular protein characterization of six Bp isolates plus the pertussis acellular vaccine strain Bp Tohama I (T), utilizing gel-free proteomics-based mass spectrometry (MS). MS/MS tryptic peptide detection and protein database searching combined with western blot analysis revealed three Bp isolates in this study had markedly reduced detection of pertactin (Prn), a subunit of pertussis acellular vaccines. Additionally, antibody affinity capture technologies were implemented using anti-Bp T rabbit polyclonal antisera and whole cellular proteins to identify putative immunogens. Proteome profiling could shed light on pathogenesis and potentially lay the foundation for reduced infection transmission strategies and improved clinical diagnostics.