Research Days 2023 - On Demand Presentations

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On Demand

Abstract

Staphylococcus aureus, a gram-positive, commensal bacterium, causes several opportunistic diseases and infections in humans. Furthermore, some strains of S.aureus have developed resistance to antibiotics such as methicillin and pose a health risk. Pattern Recognition Receptors (PRRs) on host immune cells recognize specific components of S.aureus cell wall and mount an immune response against the bacteria. One such PRR is the Nuclear Oligomerization Domain 2 (NOD2) receptor which is present in the cytosol of host immune cells. NOD2 recognizes Muramyl-Dipeptide (MDP), a component of the peptidoglycan cell wall of several bacteria including S. aureus. The binding of MDP to NOD2 allows the activation of host immune response by secretion of pro-inflammatory cytokines and antimicrobial compounds. The exact mechanism of binding of bacterial components to the NOD2 receptor is unknown. This study aims to utilize a S.aureus mutant library to screen for bacterial mutants that either fail to bind to NOD2 or show higher binding to NOD2 when compared to wild type bacteria. Commercially available Human Embryonic Kidney cells transfected with a mouse NOD2 gene (HEK-BLUE mNOD2) will be treated with wild type or mutant bacterial components from the S.aureus mutant library. Binding of bacterial components to NOD2 will be detected using a colorimetric assay and quantified by absorbance at 650nm. This screen will enable us to determine the bacterial components that are essential to NOD2 binding and may also shed light on ways by which S.aureus evades host immune responses.

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Role of NOD2 Receptor in Immune Response to Staphylococcus Aureus

On Demand

Staphylococcus aureus, a gram-positive, commensal bacterium, causes several opportunistic diseases and infections in humans. Furthermore, some strains of S.aureus have developed resistance to antibiotics such as methicillin and pose a health risk. Pattern Recognition Receptors (PRRs) on host immune cells recognize specific components of S.aureus cell wall and mount an immune response against the bacteria. One such PRR is the Nuclear Oligomerization Domain 2 (NOD2) receptor which is present in the cytosol of host immune cells. NOD2 recognizes Muramyl-Dipeptide (MDP), a component of the peptidoglycan cell wall of several bacteria including S. aureus. The binding of MDP to NOD2 allows the activation of host immune response by secretion of pro-inflammatory cytokines and antimicrobial compounds. The exact mechanism of binding of bacterial components to the NOD2 receptor is unknown. This study aims to utilize a S.aureus mutant library to screen for bacterial mutants that either fail to bind to NOD2 or show higher binding to NOD2 when compared to wild type bacteria. Commercially available Human Embryonic Kidney cells transfected with a mouse NOD2 gene (HEK-BLUE mNOD2) will be treated with wild type or mutant bacterial components from the S.aureus mutant library. Binding of bacterial components to NOD2 will be detected using a colorimetric assay and quantified by absorbance at 650nm. This screen will enable us to determine the bacterial components that are essential to NOD2 binding and may also shed light on ways by which S.aureus evades host immune responses.