Master of Science (MS)
School of Animal Science
Brucella is a genus of Gram negative, facultative intracellular pathogens which cause brucellosis, one of the most wide spread zoonotic diseases. Brucellosis causes a severe economic burden due to reproductive loss in animals and human infection. Vaccination of animals has proven to be the most effective means of controlling brucellosis; however the current live-attenuated vaccines are not considered ideal. The live-attenuated vaccines Brucella abortus Strain 19 and RB51 remain pathogenic to humans, and the former interferes with diagnostic tests due to induction of antibodies against the O-polysaccharide. DNA vaccination with single antigens has proven to be successful at protecting mice against B. abortus challenge, however this method is less effective in large animals. Immunization with a combination of antigens has been shown to provide more protection than single antigens. In an attempt to develop a better DNA vaccine, two multivalent multi-epitope plasmids were constructed using known protective antigens and bioinformatics technologies. Epitopes predicted to induce cell-mediated immunity were selected from Cu/Zn superoxide dismutase, outer membrane protein (Omp) 16, Omp19, ribosomal subunit L7/L12, BP26, ribosomal subunit L9, and Omp25. The plasmids were transfected into Chinese hamster ovary (CHO K1) cells, and PCR was used to confirm presence of the sequences in the genome. The transcription of the BabV1 and BabV2 genes to RNA was confirmed using RT-PCR. Finally, Western blots using sera from Strain 19 infected goats suggest the protein is not recognized by the humoral response of vaccinated animals. Further research is required to determine if the p425/BabV1 and p425/BabV2 vaccines are recognized by the cell-mediated immune response of infected or vaccinated animals.
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McGee, Michael, "Design and Humoral Analysis of Two Epitope-Based Brucella abortus DNA Vaccines" (2017). LSU Master's Theses. 4397.