Title page for ETD etd-01212010-182722


Type of Document Dissertation
Author Rajasekaran, Parthiban
Author's Email Address rajparthy@gmail.com
URN etd-01212010-182722
Title Development of an Antibiotic Resistance Free Bivalent Vaccine Against Swine Brucellosis and Swine Influenza
Degree PhD
Department Biomedical and Veterinary Sciences
Advisory Committee
Advisor Name Title
Boyle, Stephen M. Committee Chair
Mukhopadhyay, Biswarup Committee Member
Roberts, Paul Christopher Committee Member
Schurig, Gerhardt G. Committee Member
Sriranganathan, Nammalwar Committee Member
Keywords
  • swine influenza
  • multivalent vaccine
  • Brucella suis
  • leuB
  • leucine auxotroph
  • Brucella abortus strain RB51
Date of Defense 2009-12-09
Availability unrestricted
Abstract
Livestock across the world contract several infectious diseases of both bacterial and viral origin. Swine brucellosis caused by Brucella suis and swine influenza caused by Influenza A virus affect both domestic and feral swine populations. Both the diseases have zoonotic potential to cause disease in humans with serious complications apart from inflicting huge economic losses. Infected feral swine can also act as a source of spread and outbreak where the disease is not endemic. At present, there is no vaccine available for swine brucellosis. The currently used swine influenza vaccine may not be effective against influenza strains like the recent H1N1 strain that caused a pandemic. To develop an effective bivalent vaccine for swine against these two diseases, a leucine auxotroph of the USDA approved vaccine B. abortus strain RB51 was constructed along with leuB gene complementing plasmid pNS4 to over-express antigens from Brucella and influenza. This antibiotic resistance free system over-expressed Brucella derived antigens SOD, L7/L12 and WboA in three different constructs. Against a virulent challenge of B. suis, the candidate vaccine strain over-expressing both SOD and WboA protected mice more significantly than the control group and was also found to be better protective than other candidate vaccine strains over-expressing either SOD and L7/L12 together or SOD alone. Immunoassays (ELISA) suggested that the protection afforded is Th1 type mediated immune response, as cytokine IFN-γ and IgG2a antibody sub-isotype was observed in the splenocyte culture supernatant and serum samples respectively. The strain RB51leuB platform was not expressing influenza derived antigens Hemagglutinin (HA) and Nucleoprotein (NP) when screened for expression by immunoblot. Influenza antigens, HA, NP and ectodomain of matrix protein M2e, were not found to be expressing even after optimizing their codon usage to suit Brucella tRNA preference. However, RT-PCR showed that the influenza genes mRNA were produced. In conclusion, this dissertation describes the construction of an environmentally safe antigen over-expression platform and successful employment of the system as a candidate vaccine in protecting mice against B. suis challenge. This new platform is a potential candidate for developing vaccines against other infectious diseases of livestock. This document also discusses alternate strategies for expressing influenza antigens in a Brucella platform.
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