التفاصيل البيبلوغرافية
| العنوان: |
Identification and Evaluation of Brucella Recombinant Outer Membrane Proteins as Subunit Vaccinogen Candidates in the Mouse Model of Brucellosis |
| المؤلفون: |
Gomez, Gabriel |
| المساهمون: |
Ficht, Thomas A, Adams, Leslie G, Mwangi, Waithaka, Burghardt, Robert |
| سنة النشر: |
2013 |
| المجموعة: |
Texas A&M University Digital Repository |
| مصطلحات موضوعية: |
Brucella, outer membrane proteins, vaccine, antigens, reverse vaccinology, mouse |
| الوصف: |
Despite being amongst the most common zoonotic diseases in the world, brucellosis is a neglected disease for which an approved vaccine for human use does not exist. Thus far, the traditional approaches to Brucella antigen selection for subunit vaccine development have yielded unacceptable results. In this work, we evaluated the predictive ability of a multistep Brucella antigen selection process with in vitro immunological and invasion assays and in vivo protection experiments. Initial in silico screening for antigens was performed via genomic sequence analysis where 27 Brucella melitensis open reading frames (ORF) coding for outer membrane proteins bearing MHC epitopes, adhesin and conserved properties were identified. Evidence for a role in any aspect of Brucella virulence (i.e., invasion, co-regulation/expression with known Brucella virulence factors, intracellular adaptation) was then used to narrow the list of candidate antigens. To further increase confidence in the candidate ORF putative role in Brucella pathogenesis, differential expression of candidate ORF was evaluated using previously generated global transcriptomics data in in vitro HeLa and in vivo bovine models of acute Brucella infection. Protein expression in the E. coli heterologous system resulted in the successful expression of OmpW, BtuB, Omp22, Hia, and FlgK. With regards to virulence, the two proteins with the highest predicted adhesin scores conferred an invasive phenotype to the non-invasive BL-21 E. coli strain in alveolar epithelial cells. From an immunogenicity standpoint, all proteins elicited IgG production in Brucella-exposed goats, mouse and humans. Antigen-specific recall responses in splenocytes from C57BL/6 mice immunized with a cocktail of the three proteins with highest MHC scores revealed a mixed Th1/Th2 response with a comparatively greater Th1 response. In protection studies, subcutaneous (SQ) immunization with BtuB, Hia and FlgK, individually, promoted bacterial clearance following a robust intraperitoneal challenge dose ... |
| نوع الوثيقة: |
thesis |
| وصف الملف: |
application/pdf |
| اللغة: |
unknown |
| العلاقة: |
https://hdl.handle.net/1969.1/149282Test |
| الإتاحة: |
https://hdl.handle.net/1969.1/149282Test |
| رقم الانضمام: |
edsbas.4E867205 |
| قاعدة البيانات: |
BASE |
