Year

2015

Season

Spring

Paper Type

Master's Thesis

College

College of Arts and Sciences

Degree Name

Master of Science in Biology (MS)

Department

Biology

NACO controlled Corporate Body

University of North Florida. Department of Biology

First Advisor

Dr. Terri N. Ellis

Second Advisor

Dr. Judith Ochrietor

Rights Statement

http://rightsstatements.org/vocab/InC/1.0/

Third Advisor

Dr. Gregory Ahearn

Department Chair

Dr. Daniel Moon

College Dean

Dr. Barbara Hetrick

Abstract

Klebsiella pneumoniae is a Gram-negative pathogen associated with numerous infections. Like all Gram-negative bacteria, K. pneumoniae naturally release outer membrane vesicles (OMVs) during all stages of cellular growth. OMVs are composed of the outer membrane components such as lipopolysaccharide (LPS) and outer membrane proteins and contain cytosolic and periplasmic proteins in the lumen. K. pneumoniae is often found to lack an O antigen. The absence of the O antigen has been reported to alter the protein content of the membrane which may further alter the immune response elicited by K. pneumoniae. Therefore the purpose of this study was to analyze the protein content and inflammatory responses of the cellular outer membrane (OM) and the secreted OMVs. A secondary goal of this study was to evaluate any changes to the membrane protein content due to O antigen loss and determine if the loss of the O antigen influences the inflammatory response. This study demonstrated that the cellular OM and OMVs have distinct protein profiles as well as inflammatory responses. OMVs were highly enriched in outer membrane proteins compared to the cellular OM and had a significantly stronger inflammatory response. The protein content of the OM and OMVs were further modified by the absence of the O antigen from the outer membrane. Although there were no significant differences seen in the wild type and O antigen deficient OM inflammatory responses, the loss of the O antigen resulted in OMVs that produced a significantly stronger IL-6 response.

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