Proteome analyses of Bordetella pertussis & B. parapertussis, the causative agents of whooping cough, a re-emerging disease

ALVARES HAYES, JIMENA; GORGOJO, JUAN PABLO; HOLUBOVA, J; SCHMIDT, F; SURMANN, KRISTIN; CAFIERO, JUAN HILARIO; VALDEZ, HUGO; DEPKE, M; RODRIGUEZ, MARÍA EUGENIA; LAMBERTI, YANINA; OVIEDO, JUAN MARCOS; VECEREK, B; DHOPLE, VISHNU M.; VÖLKER, UWE

Leipzig

Conferencia; 6th Joint Conference of the DGHM & VAAM 2020. 72nd Annual Meeting of the German Society for Hygiene and Microbiology (DGHM), Annual Meeting 2020 of the Association for General and Applied Microbiology (VAAM).; 2020

DGHM & VAAM

Proteome analyses of Bordetella pertussis & B. parapertussis, the causative agents of whooping cough, a re-emerging disease Kristin Surmann1, Yanina Lamberti2, Jimena Alvarez Hayes2, Juan Hilario Cafiero2, Juan Marco Oviedo2, Juan Gorgojo2, Hugo Valdez2, Branislav Vecerek3, Jana Holubova3, Maren Depke1, Vishnu M. Dhople1, Frank Schmidt1, Maria Eugenia Rodriguez2, Uwe Völker1 1 Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Greifswald, Germany2 CINDEFI (UNLP CONICET La Plata), Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Argentina3 Laboratory of Molecular Biology of Bacterial Pathogens, Institute of Microbiology of the ASCR, v.v.i., Prague, Czech RepublicIntroductionEven in fully vaccinating countries, whooping cough caused by Bordetella pertussis (Bp) and B. parapertussis (Bpp), is regarded as re-emerging disease. Besides vaccine inefficiency, intracellular survival of the pathogens leads to dissemination amongst the population. ObjectivesIn order to elucidate bacterial adaptation to infection related conditions, we have analysed the global proteome of both pathogens to iron-limitation and additionally, upon internalization of Bp. Materials & methodsBp Tohama I, an isogenic Δhfq mutant, and Bpp CN2591 were cultivated in iron-replete and iron-depleted Stainer-Scholte medium until late exponential growth phase. Bacterial proteomes were analysed by nanoLC-MS/MS. In addition, human monocytic THP-1 cells were infected with Bp Tohama I (MOI 150) and after 2 h, non-internalized bacteria were killed with polymyxin B sulfate. The proteome of recovered intracellular Bp was examined 3 h and 48 h post infection and compared to extracellular Bp grown under similar conditions. In addition, infection was repeated with mutants in BP0414, encoding a MgtC homologue or hfq to assess intracellular survival by bacterial counting. ResultsThe chaperone Hfq modulates 33% of Bp proteins regulated during iron limitation (Alvarez Hayes et al., J Proteomics, 2019) and influenced bacterial survival. Upon internalization, 40% of the detected proteins showed altered levels including proteins involved in stress response, iron uptake, metabolism, and virulence (Lamberti et al., J Proteomics, 2016). A follow-up study revealed that MgtC plays a role in the adaptation of Bp to the mildly acidic conditions inside phagosomes (Cafiero et al., PLoS One, 2018). Moreover, we identified proteins of stress resistance and virulence exclusively in Bpp that might help explaining differences in pathogenesis of both species (Oviedo et al., J Proteomics, 2019). ConclusionHere, we present comprehensive data of the pertussis causing agents and help to understand their pathogenicity and pave the way for follow-up studies of the infected host by proteomics and immunoproteomics.