澳大利亚流行博德特氏菌表面组分析揭示潜在疫苗抗原。
Surfaceome analysis of Australian epidemic Bordetella pertussis reveals potential vaccine antigens.
机构信息
School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales, Australia.
Bioanalytical Mass Spectrometry Facility, University of New South Wales, Sydney, New South Wales, Australia.
出版信息
Vaccine. 2020 Jan 16;38(3):539-548. doi: 10.1016/j.vaccine.2019.10.062. Epub 2019 Nov 6.
Since acellular vaccines (ACV) were introduced in Australia, epidemic Bordetella pertussis strains changed from single nucleotide polymorphism (SNP) cluster II to SNP cluster I. Our previous proteomic analysis identified potential proteomic adaptations in the whole cell and secretome of SNP cluster I. Additionally, current ACVs were shown to be less efficacious against cluster I in mice models and there is a pressing need to discover new antigens to improve the ACV. One important source of novel antigens is the surfaceome. Therefore, in this study we established surface shaving in B. pertussis to compare the surfaceome of SNP cluster I (L1423) and II (L1191), and identify novel surface antigens for vaccine development. Surface shaving using 1 μg of trypsin for 5 min identified 126 proteins with the most abundant being virulence-associated and known outer membrane proteins. Cell viability counts showed minimal lysis from shaving. The proportion of immunogenic proteins was higher in the surfaceome than in the whole cell and secretome. Key differences in the surfaceome were identified between SNP cluster I and II, consistent with those identified in the whole cell proteome and secretome. These differences include unique transport proteins and decreased immunogenic proteins in L1423, and provides further evidence of proteomic adaptation in SNP cluster I. Finally, a comparison of proteins in each sub-proteome identified 22 common proteins. These included 11 virulence proteins (Prn, PtxA, FhaB, CyaA, TcfA, SphB1, Vag8, BrkA, BopD, Bsp22 and BipA) and 11 housekeeping proteins (TuF, CtpA, TsF, OmpH, GltA, SucC, SucD, FusA, GroEL, BP3330 and BP3561) which were immunogenic, essential and consistently expressed thus demonstrating their potential as future targets. This study established surface shaving in B. pertussis, confirmed key expression differences and identified unknown surface proteins which may be potential vaccine antigens.
自从无细胞疫苗（ACV）在澳大利亚推出以来，流行的百日咳博德特氏菌菌株已从单核苷酸多态性（SNP）簇 II 转变为 SNP 簇 I。我们之前的蛋白质组学分析确定了 SNP 簇 I 全细胞和分泌组中潜在的蛋白质组适应性。此外，目前的 ACV 在小鼠模型中对簇 I 的效果较差，因此迫切需要发现新的抗原来改进 ACV。新抗原的一个重要来源是表面组。因此，在这项研究中，我们在百日咳博德特氏菌中建立了表面刮除，以比较 SNP 簇 I（L1423）和 II（L1191）的表面组，并确定用于疫苗开发的新型表面抗原。使用 1μg 的胰蛋白酶刮除 5 分钟，鉴定出 126 种蛋白质，其中最丰富的是与毒力相关的和已知的外膜蛋白。细胞活力计数显示刮除后几乎没有裂解。表面组中的免疫原性蛋白比例高于全细胞和分泌组。在 SNP 簇 I 和 II 之间鉴定到表面组的关键差异，这与全细胞蛋白质组和分泌组中的差异一致。这些差异包括 SNP 簇 I 中独特的转运蛋白和免疫原性蛋白减少，这进一步证明了 SNP 簇 I 中的蛋白质组适应性。最后，对每个亚蛋白质组中的蛋白质进行比较，鉴定出 22 种共同蛋白质。这些蛋白质包括 11 种毒力蛋白（Prn、PtxA、FhaB、CyaA、TcfA、SphB1、Vag8、BrkA、BopD、Bsp22 和 BipA）和 11 种管家蛋白（TuF、CtpA、TsF、OmpH、GltA、SucC、SucD、FusA、GroEL、BP3330 和 BP3561），这些蛋白具有免疫原性、必需性和一致性表达，因此证明了它们作为未来靶标的潜力。本研究在百日咳博德特氏菌中建立了表面刮除，证实了关键的表达差异，并鉴定了未知的表面蛋白，这些蛋白可能是潜在的疫苗抗原。
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