ABSTRACT: Deciphering the genetic basis of Moraxella catarrhalis complement resistance: Gene expression analysis of Moraxella catarrhalis BBH18 upon exposure to normal human serum.
Project description:The complement system is an important part of the innate defense against invading pathogens. The ability to resist complement-mediated killing is considered to be an important virulence trait for the human-restricted respiratory tract pathogen M. catarrhalis, as most disease-associated M. catarrhalis isolates are complement-resistant. Here we studied the molecular basis of M. catarrhalis complement-resistance by transcriptome profiling upon exposure to 10% normal human serum (NHS).
Project description:The complement system is an important part of the innate defense against invading pathogens. The ability to resist complement-mediated killing is considered to be an important virulence trait for the human-restricted respiratory tract pathogen M. catarrhalis, as most disease-associated M. catarrhalis isolates are complement-resistant. Here we studied the molecular basis of M. catarrhalis complement-resistance by transcriptome profiling upon exposure to 10% normal human serum (NHS). After 1h exposure of M. catarrhalis BBH18 to 10% NHS (n = 5) or the NHS dilution buffer alone (control, n = 5), total RNA was isolated and labeled cDNA was generated according to standard Nimblegen gene expression array protocols and hybridized to 4x72K Nimblegen M. catarrhalis expression arrays for read-out.
Project description:Comparison of the gene expression profile of Moraxella catarrhalis grown in the presence of 20% pooled human sputum in chemically-defined medium relative to Moraxella catarrhalis grown in chemically-defined medium alone.
Project description:Comparison of the gene expression profile of Moraxella catarrhalis grown in the presence of 20% pooled human sputum in chemically-defined medium relative to Moraxella catarrhalis grown in chemically-defined medium alone. Moraxella catarrhalis ATCC43617 was grown to mid-logarithmic phase either in the presence of 20% pooled human sputum in chemically-defined medium or in chemically-defined medium alone. Total RNA was extracted from bacterial cells exposed to each of these conditions and cDNA was generated for CyDye labelling. 3 biologic replicates were generated and each replicate underwent a dye swap (total of 6 experimental data collections). The gene expression profile reported is that of Moraxella catarrhalis grown in the presence of pooled human sputum in a chemically-defined medium relative to Moraxella catarrhalis grown only in the presence of the chemically-defined medium.
Project description:During the course of infection, respiratory pathogens like Moraxella catarrhalis needs to adhere to epithelial cells of different host niches such as the nasopharynx and lungs. Consequently, efficient adhesion to epithelial cells is considered an important virulence trait of M. catarrhalis. We examined the interaction between human pharyngeal epithelial Detroit 562 cells and M. catarrhalis BBH18 during adherence using a combination of Tn-seq, a genome-wide negative selection screenings technology, and expression profiling of both host and pathogen. The results described in this study are further discussed in Stefan P.W. de Vries, Marc J. Eleveld, Peter W.M. Hermans, Hester J. Bootsma: Characterization of the molecular interplay between Moraxella catarrhalis and human respiratory tract epithelial cells, submitted.
Project description:During the course of infection, respiratory pathogens like Moraxella catarrhalis needs to adhere to epithelial cells of different host niches such as the nasopharynx and lungs. Consequently, efficient adhesion to epithelial cells is considered an important virulence trait of M. catarrhalis. We examined the interaction between human pharyngeal epithelial Detroit 562 cells and M. catarrhalis BBH18 during adherence using a combination of Tn-seq, a genome-wide negative selection screenings technology, and expression profiling of both host and pathogen. The results described in this study are further discussed in Stefan P.W. de Vries, Marc J. Eleveld, Peter W.M. Hermans, Hester J. Bootsma: Characterization of the molecular interplay between Moraxella catarrhalis and human respiratory tract epithelial cells, submitted.
Project description:RNA-sequencing of uninfected and Moraxella catarrhalis-infected bone marrow-derived macrophages (BMDMs) isolated from Ifnar1 knockout and wild-type mice.
