Title page for ETD etd-07012010-214035


Type of Document Master's Thesis
Author Cassidy, Clifton Clark
Author's Email Address cassidy@vt.edu
URN etd-07012010-214035
Title Brucella abortus Strain RB51 Outer Membrane Vesicles as a Vaccine Against Brucellosis in a Murine Model
Degree Master of Science
Department Veterinary Medical Sciences
Advisory Committee
Advisor Name Title
Boyle, Stephen M. Committee Chair
Sriranganathan, Nammalwar Committee Co-Chair
Mullarky, Isis K. Committee Member
Keywords
  • outer membrane vesicles
  • strain RB51
  • Brucella abortus
  • adjuvant
  • blebs
  • vaccine
  • mice
  • OMV
  • malaria
  • FRAP
Date of Defense 2010-02-03
Availability unrestricted
Abstract
Brucella abortus is a zoonotic agent that primarily infects cattle and causes brucellosis. B. abortus strain RB51 is a live, attenuated vaccine licensed for cattle. However, there is no available vaccine to prevent human brucellosis. Outer membrane vesicles have been tested as potential vaccines to prevent diseases caused by bacterial species. OMV are constantly released from Gram-negative bacteria. They are comprised principally of the outer membrane components and periplasmic proteins from the bacterial cell envelope. The research in this thesis examined the adjuvant property of non-replicative, metabolically active irradiated strain RB51 and the protective ability of OMV derived from strain RB51. Irradiated B. abortus strain RB51 was assessed for its ability to act as an adjuvant to induce protection against malaria. It was found that irradiated B. abortus strain RB51 administered along with fasciclin related adhesive protein (FRAP) to mice induced a protective immune response and a significant decrease in parasitemia after challenge with Plasmodium berghei. Strain RB51 and strain RB51 over-producing Cu/Zn superoxide dismutase (Cu/Zn SOD) were used to produce OMV. Western blotting and SDS-PAGE gel staining confirmed the presence of OMV and the over-production of Cu/Zn SOD. OMV were delivered to mice using an intraperitoneal route and, in some cases, with aluminum hydroxide adjuvant. The immune response was assessed by antibody isotyping with respect to OMV and measuring splenic clearance (i.e. protection) from a B. abortus strain 2308 challenge. The results demonstrate that OMV from B. abortus strain RB51 or strain RB51 over producing Cu/Zn SOD produced a Th1 polarized immune response as measured by specific OMV antibodies and cytokines but no statistically significant protection was observed.
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