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.
| Identifer | oai:union.ndltd.org:LSU/oai:etd.lsu.edu:etd-04102017-085926 |
| Date | 27 April 2017 |
| Creators | McGee, Michael |
| Contributors | Cooper, Richard, Williams, Cathleen, Elzer, Phil |
| Publisher | LSU |
| Source Sets | Louisiana State University |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.lsu.edu/docs/available/etd-04102017-085926/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached herein a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to LSU or its agents the non-exclusive license to archive and make accessible, under the conditions specified below and in appropriate University policies, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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