Optimization of an Immune Assay to Detect Binding of NOD2 to Staphylococcus Aureus

<p><em>Staphylococcus aureus</em>, a gram-positive, commensal bacterium, is the cause of a multitude of opportunistic diseases and infections. While <em>S. aureus</em> has evolved ways to escape innate immune responses in the host, the mechanisms are not fully understood. One area of active research involves mechanisms by which <em>S. aureus</em> avoids detection by Pattern Recognition Receptors (PRRs), such as Nuclear Oligomerization Domain 2 (NOD2), present on the surface of and inside innate immune cells. This study aimed to design and optimize an assay that can be utilized to investigate <em>S. aureus</em> components that contribute to the activation of NOD2 receptors. The assay was designed using one wild type and two bacterial strains from the Nebraska Transposon Mutant Library (NTML), murine NOD2 transgenic cells, and two detection systems. I hypothesized that both mutant strains would have differential binding to NOD2 compared to the wild-type strain. Results showed that only one mutant strain, USA300_ 1095, which codes for the <em>carA </em>gene showed differential binding with our detection protocols. While this study focused on optimizing the assay, the protocols created can be expanded to the entire NTML to identify novel <em>S. aureus</em> genes involved in the NOD2 binding pathway.</p>

Name thesis:degree_name

Master of Science

Level thesis:degree_level

Thesis

Discipline thesis:degree_discipline

Biology

Year dc:date.available

2024

Author dc:creator

• McKenzie, Precious

Contributors dc:contributor

• Timothy Gsell
• Aparna Palakodeti
• Walter Henne

• Biology
• Immunology and Infectious Disease
• Microbiology

Repository record dc:identifier

https://opus.govst.edu/theses/155

OAI identifier oai:identifier

oai:opus.govst.edu:theses-1154