Metabolic pathways of Vibrio cholerae and their impact on the transmission fitness of the bacterial pathogen
S. Schild studies molecular mechanisms of bacterial pathogenesis and bacterial adaptation in response to environmental and host conditions. Surface structures and outer membrane vesicles (OMVs) of Gram-negative bacteria are emphasized including their impact on virulence and vaccine development. Selected model organisms include Vibrio cholerae and Haemophilus influenzae, representing the causative agents of cholera or upper respiratory tract infections, otitis media and meningitis, respectively. S. Schild holds two patents on OMVs as vaccine candidates for Gram-negative bacteria. Current research focuses on understanding OMV secretion and sorting mechanisms using H. influenzae as a model organism. A second research focus investigates variation in gene regulation throughout the lifecycle of the facultative human pathogen V. cholerae, which is marked by recurrent transitions between the aquatic environment and the human host. Among the genes subject to variable regulation were several enzymes important to second messenger cascades, protein glycosylation as well as to phosphate metabolism and storage processes. All of these seem to be important for several stages in the lifecycle of V. cholerae, including biofilm formation, survival in the aquatic reservoir, virulence and transmission of cholera. DK students will pursue a mechanistic understanding of candidate enzymes and their cellular functions in Funding Period 4. S. Schild’s laboratory collaborates with several groups within and outside the DK to share expertise on animal models and to develop suitable in vivo and colonization studies for other bacterial pathogens.
Laboratory know-how and infrastructureThe project will be carried out in the biohazard class 2 labortatories at the Institute of Molecular Biosciences. All necessary equipment such as bacterial biohazard class 2 facilities equipped with typical laboratory instruments, state-of-the-art animal facility with IVC (individually ventilated cages)-system, cell culture facilities, flow benches, ultracentrifuges, French press, horizontal and vertical electrophoresis facilities, plate reader, qRT-PCR, and epifluorescence microscopes will be provided by the department. The applicant’s laboratory has comprehensive experience handling and genetically manipulating pathogenic bacteria and established a variety of in vitro and in vivo assays to study bacterial adaptation processes. Routine procedures include gene cloning, insertion and deletion mutagenesis, protein expression, subcellular fractionation, enzyme kinetic assays, immunoblot analysis, ELISA, biofilm assays, various transcriptional reporter assays and pathogen – host interaction assays using cell culture as well as in vivo models. S. Schild has been trained in special courses on laboratory animals (according to the recommendations of the Federation of European Laboratory Animal Science Associations, FELASA), which are certified by GV-Solas. He has comprehensive experience with animal experiments including colonization and immunization studies.