Harth-Chu Erika N, Alves Lívia A, Theobaldo Jéssica D, Salomão Mariana F, Höfling José F, King William F, Smith Daniel J, Mattos-Graner Renata O
Department of Oral Diagnosis, Piracicaba Dental School, UNICAMP, Piracicaba, Brazil.
Department of Immunology and Infectious Disease, The Forsyth Institute, Cambridge, MA, United States.
Front Microbiol. 2019 Nov 12;10:2567. doi: 10.3389/fmicb.2019.02567. eCollection 2019.
is an abundant member of the commensal microbiota of the oral cavity and pharynx, which has the potential to promote systemic infections. By analyzing a collection of strains isolated from the oral cavity at commensal states or from systemic infections (blood strains), we established that ubiquitously express the surface immunodominant protein, PcsB (also called GbpB), required for binding to sucrose-derived exopolysaccharides (EPS). Immuno dot blot assays with anti-PcsB antibodies and RT-qPCR transcription analyses revealed strain-specific profiles of PcsB production associated with diversity in transcriptional activities. Additionally, blood strains showed significantly higher levels of PcsB expression compared to commensal isolates. Because co-colonizes dental biofilms, and secretes glucosyltransferases (GtfB/C/D) for the synthesis of highly insoluble EPS from sucrose, profiles of binding to EPS, biofilm formation and evasion of the complement system were assessed in sucrose-containing BHI medium supplemented or not with filter-sterilized culture supernatants. These analyses showed significant binding to EPS and biofilm formation in the presence of supernatants supplemented with sucrose, compared to BHI or BHI-sucrose medium. In addition, these phenotypes were abolished if strains were grown in culture supernatants of a -defective mutant. Importantly, GtfB/C/D-associated phenotypes were enhanced in high PcsB-expressing strains, compared to low PcsB producers. Increased PcsB expression was further correlated with increased resistance to deposition of C3b/iC3b of the complement system after exposure to human serum, when strains were previously grown in the presence of supernatants. Finally, analyses of PcsB polymorphisms and bioinformatic prediction of epitopes with significant binding to MHC class II alleles revealed that blood isolates harbor PcsB polymorphisms in its functionally conserved CHAP-domain, suggesting antigenic variation. These findings reveal important roles of PcsB in -host interactions under commensal and pathogenic states, highlighting the need for studies to elucidate mechanisms regulating PcsB expression in this species.
是口腔和咽部共生微生物群的丰富成员,有可能促进全身感染。通过分析从共生状态的口腔或全身感染中分离的菌株(血液菌株),我们确定 普遍表达与蔗糖衍生的胞外多糖(EPS)结合所需的表面免疫显性蛋白PcsB(也称为GbpB)。用抗PcsB抗体进行的免疫斑点印迹分析和RT-qPCR转录分析揭示了与 转录活性多样性相关的PcsB产生的菌株特异性谱。此外,与共生分离株相比,血液菌株显示出显著更高水平的PcsB表达。由于 共同定殖于牙菌斑,并分泌葡糖基转移酶(GtfB/C/D)以从蔗糖合成高度不溶性的EPS,因此在补充或不补充经滤膜除菌的 培养上清液的含蔗糖BHI培养基中评估了 与EPS结合、生物膜形成和补体系统逃避的情况。这些分析表明,与BHI或BHI-蔗糖培养基相比,在补充有蔗糖的 上清液存在下, 与EPS的结合和生物膜形成显著。此外,如果菌株在 缺陷突变体的培养上清液中生长,这些表型会被消除。重要的是,与低PcsB产生者相比,在高PcsB表达菌株中,GtfB/C/D相关表型增强。当菌株先前在 上清液存在下生长时,PcsB表达的增加进一步与暴露于人血清后对补体系统C3b/iC3b沉积的抗性增加相关。最后,对PcsB多态性的分析和与MHC II类等位基因有显著结合性的表位的生物信息学预测表明,血液分离株在其功能保守的CHAP结构域中存在PcsB多态性,提示存在抗原变异。这些发现揭示了PcsB在共生和致病状态下与宿主相互作用中的重要作用,突出了阐明该物种中调节PcsB表达机制的研究的必要性。