Defense Date

2009

Document Type

Thesis

Degree Name

Master of Science

Department

Physiology and Biophysics

First Advisor

Janina P. Lewis

Abstract

Periodontal disease is one of the most prevalent human diseases affecting millions of individuals. Bacteria are the trigger of the host response that then results in initiation and progression of periodontal disease. The interactions between the host and the bacteria, although very complex and intricate, are not well understood. Species of gram-negative anaerobic bacteria implicated in the development of periodontal disease include Porphyromonas gingivalis and Prevotella intermedia. The goal of our study was to identify P. intermedia surface proteins interacting with TLR2 receptor. Using Cy5-labeled surface proteins we identified multiple proteins using our capture assay. The interaction of three of those proteins was further characterized using recombinant forms of those proteins. Using ELISA assay, we confirmed the TLR2-binding function of our selected recombinant proteins at various protein amounts. Also, we confirmed the ability to activate TLR2 signaling by challenging HEK cells overexpressing TLR2 with our recombinant proteins and measuring the SEAP (secreted alkaline phosphatase) activity which is a reporter gene that is under the control of an NF-κB inducible promoter. Our results suggest that P. intermedia 17 Pin_A0326 and Pin_A0102 proteins play a role in initiating the innate immune response by binding to TLR2. The protein encoded by a 2101-3171bp segment of Pin _A0326 shows the highest degree of signaling activity in our HEK cells study. Also, similar results were observed using Pin_ A0324-encoded protein to challenge the HEK cells. These results suggest that these P. intermedia surface proteins play a role in interaction of the bacterium with host cells.

Comments

Scanned, with permission from the author, from the original print version, which resides in University Archives.

Rights

© The Author

Is Part Of

VCU University Archives

Is Part Of

VCU Theses and Dissertations

Date of Submission

10-13-2021

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