Department of Genetics and Genome Biology, University of Leicester, Leicester, United Kingdom.
Department of Mathematics, University of Leicester, Leicester, United Kingdom.
PLoS One. 2018 May 30;13(5):e0197186. doi: 10.1371/journal.pone.0197186. eCollection 2018.
Factor H binding protein (fHbp) is a major protective antigen in 4C-MenB (Bexsero®) and Trumenba®, two serogroup B meningococcal vaccines, wherein expression level is a determinant of protection. Examination of promoter-containing intergenic region (IGR) sequences indicated that nine fHbp IGR alleles covered 92% of 1,032 invasive meningococcal strains with variant 1 fHbp alleles. Relative expression values for fHbp were determined for 79 meningococcal isolates covering ten IGR alleles by quantitative reverse transcriptase polymerase chain reaction (qRT PCR). Derivation of expression clusters of IGR sequences by linear regression identified five expression clusters with five nucleotides and one insertion showing statistically associations with differences in expression level. Sequence analysis of 273 isolates examined by the Meningococcal Antigen Typing Scheme, a sandwich ELISA, found that coverage depended on the IGR expression cluster and vaccine peptide homology combination. Specific fHbp peptide-IGR expression cluster combinations were designated as 'at risk' for coverage by 4C-MenB and were detected in multiple invasive meningococcal disease cases confirmed by PCR alone and occurring in partially-vaccinated infants. We conclude that sequence-based analysis of IGR sequences is informative for assessing protein expression and has utility for culture-independent assessments of strain coverage by protein-based vaccines.
因子 H 结合蛋白(fHbp)是 4C-MenB(Bexsero®)和 Trumenba®两种 B 群脑膜炎球菌疫苗的主要保护性抗原,其表达水平是保护效果的决定因素。对包含启动子的基因间区(IGR)序列的检查表明,9 种 fHbp IGR 等位基因涵盖了 1032 株侵袭性脑膜炎球菌菌株中的 92%,其中具有变异 1 fHbp 等位基因。通过定量逆转录聚合酶链反应(qRT-PCR)对覆盖十个 IGR 等位基因的 79 株脑膜炎球菌分离株的 fHbp 相对表达值进行了测定。通过线性回归推导出 IGR 序列的表达簇,发现五个表达簇与五个核苷酸和一个插入具有统计学关联,与表达水平的差异有关。通过夹心 ELISA 对 273 株经脑膜炎球菌抗原分型方案检查的分离株进行的序列分析发现,覆盖率取决于 IGR 表达簇和疫苗肽同源性组合。特定的 fHbp 肽-IGR 表达簇组合被指定为 4C-MenB 覆盖率的“风险”,并且在仅通过 PCR 确认的多例侵袭性脑膜炎球菌病病例中检测到,并且在部分接种婴儿中也存在。我们得出结论,基于序列的 IGR 序列分析可用于评估蛋白质表达,并可用于基于蛋白质的疫苗对菌株覆盖率进行非培养评估。