Project description:Methicillin-resistant Staphylococcus aureus (MRSA) is problematic both in hospitals and the community. Currently, we have limited understanding of mechanisms of innate immune evasion used by S. aureus. To that end, we created an isogenic deletion mutant in strain MW2 (USA400) of the saeR/S two-component gene regulatory system and studied its role in mouse models of pathogenesis and during human neutrophil interaction. In this study, we demonstrate saeR/S plays a distinct role in S. aureus pathogenesis and is vital for virulence of MW2 in a mouse model of sepsis. Moreover, deletion of saeR/S significantly impaired survival of MW2 in human blood and after neutrophil phagocytosis. Microarray analysis of genes influenced by saeR/S demonstrated SaeR/S of MW2 influences a wide variety of genes with diverse biological functions. These data shed new insight into how virulence is regulated in S. aureus and associates a specific staphylococcal gene-regulatory system with invasive staphylococcal disease.
Project description:Methicillin-resistant Staphylococcus aureus (MRSA) is problematic both in hospitals and the community. Currently, we have limited understanding of mechanisms of innate immune evasion used by S. aureus. To that end, we created an isogenic deletion mutant in strain MW2 (USA400) of the saeR/S two-component gene regulatory system and studied its role in mouse models of pathogenesis and during human neutrophil interaction. In this study, we demonstrate saeR/S plays a distinct role in S. aureus pathogenesis and is vital for virulence of MW2 in a mouse model of sepsis. Moreover, deletion of saeR/S significantly impaired survival of MW2 in human blood and after neutrophil phagocytosis. Microarray analysis of genes influenced by saeR/S demonstrated SaeR/S of MW2 influences a wide variety of genes with diverse biological functions. These data shed new insight into how virulence is regulated in S. aureus and associates a specific staphylococcal gene-regulatory system with invasive staphylococcal disease. Wild type control vs mutant at two different growth phases
Project description:Staphylococcus aureus strain MW2 was exposed to the following neutrophil microbicides, hydrogen peroxide (H2O2), hypochlorous acid (HOCl) and azurophilic granule proteins. At the indicated time points, bacterial cultures were centrifuged and bacteria were lysed with RLT buffer (Qiagen) using a FastPrep system. Purification of MW2 RNA and subsequent preparation of labeled cDNA target was performed as described in Methods. Labeling of samples, GeneChip hybridization and scanning were performed according to standard Affymetrix protocols. Experiments were performed in triplicate, using three bacterial cultures from separate days for each treatment. These analyses provide an enhanced view of the mechanisms used by CA-MRSA to circumvent destruction by the human innate immune system. Keywords: time course with three treatments
Project description:Effect of the presence of Lactococcus lactis on Staphylococcus aureus transcriptome in cheese matrix. S. aureus was co-cultured with L. lactis LD61 in cheese matrix during 7 days. RNA samples were extracted at different time points (6 h, 8 h, 10 h, 24 h and 7 days) in order to monitor the dynamic response of S. aureus MW2 in cheese matrix in presence of L. lactis
Project description:Staphylococcus aureus strain MW2 was exposed to the following neutrophil microbicides, hydrogen peroxide (H2O2), hypochlorous acid (HOCl) and azurophilic granule proteins. At the indicated time points, bacterial cultures were centrifuged and bacteria were lysed with RLT buffer (Qiagen) using a FastPrep system. Purification of MW2 RNA and subsequent preparation of labeled cDNA target was performed as described in Methods. Labeling of samples, GeneChip hybridization and scanning were performed according to standard Affymetrix protocols. Experiments were performed in triplicate, using three bacterial cultures from separate days for each treatment. These analyses provide an enhanced view of the mechanisms used by CA-MRSA to circumvent destruction by the human innate immune system. 48 samples total: MW2 with the following treatments, untreated, GP (azurophilic granule proteins), H2O2 (hydrogen peroxide) or HOCl (hypochlorous acid) at 4 time points in triplicate
Project description:Staphylococcus aureus is one of the most important pathogens in humans and animals, multiply resistant strains are increasingly widespread, new agents are needed for the treatment of S. aureus. Rhein, a natural plant product, has potential antimicrobial activity against Staphylococcus aureus. We employed Affymetrix Staphylococcus aureus GeneChipsTM arrays to investigate the global transcriptional profiling of Staphylococcus aureus ATCC25923 treated with rhein. Results provided insight into mechanisms involved in rhein - Staphylococcus aureus interactions. Keywords: rhein response