Identification and immunogenic potential of B cell epitopes of outer membrane protein OmpF of Aeromonas hydrophila in translational fusion with a carrier protein

Verfasser / Beitragende:
[Mahima Sharma, Aparna Dixit]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/15(2015-08-01), 6277-6291
Format:
Artikel (online)
ID: 605506779
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024 7 0 |a 10.1007/s00253-015-6398-3  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-015-6398-3 
245 0 0 |a Identification and immunogenic potential of B cell epitopes of outer membrane protein OmpF of Aeromonas hydrophila in translational fusion with a carrier protein  |h [Elektronische Daten]  |c [Mahima Sharma, Aparna Dixit] 
520 3 |a Aeromonas hydrophila, a ubiquitous and virulent bacterial pathogen, affects a variety of fishes, including Labeo rohita. Existing treatment strategies comprise antibiotic therapies and attenuated bacterial strain-based vaccines. No functional subunit vaccine has been available until now. Given their key role in determining pathogenicity, outer membrane proteins have been successfully explored as potential vaccine candidates. We have devised a direct strategy for eliminating non-specific responses by selectively aiming the immune response against specific immunodominant epitopes of the outer membrane protein F (OmpF) of A. hydrophila (AhOmpF). Five putative epitopes of AhOmpF predicted in silico were genetically conjugated with heat labile enterotoxin chain B of E. coli (LTB). Recombinant fusion proteins expressed in E. coli were purified from solubilized inclusion bodies and refolded. The fusion protein retained GM1 ganglioside receptor binding activity of LTB, indicating proper folding. Four of the five fusion proteins were found to be highly immunogenic. Of the four proteins, antisera against the fusion protein (anti-rEpiF1) harboring 66-80 amino acid residues of the OmpF gave maximum cross-reactivity with the targeted rOmpF in enzyme-linked immunosorbent assay (ELISA) and was able to recognize both fusion partners—rOmpF and rLTB—in Western blot. Antibody isotyping of the antisera and cytokine array analysis of the culture supernatants of splenocytes from sensitized mice manifested a mixed Th1/Th2 immune response with a bias toward Th2. Anti-rEpiF1 antibodies were able to bind to the cell membrane of live A. hydrophila cells and agglutinate them. Our results thus suggest that the OmpF epitope (66-80) in fusion with a carrier protein is a promising vaccine candidate against A. hydrophila. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a Aeromonas hydrophila  |2 nationallicence 
690 7 |a OmpF  |2 nationallicence 
690 7 |a Epitope  |2 nationallicence 
690 7 |a LTB  |2 nationallicence 
690 7 |a Fusion protein  |2 nationallicence 
700 1 |a Sharma  |D Mahima  |u Gene Regulation Laboratory, School of Biotechnology, Jawaharlal Nehru University, 110067, New Delhi, India  |4 aut 
700 1 |a Dixit  |D Aparna  |u Gene Regulation Laboratory, School of Biotechnology, Jawaharlal Nehru University, 110067, New Delhi, India  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/15(2015-08-01), 6277-6291  |x 0175-7598  |q 99:15<6277  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6398-3  |q text/html  |z Onlinezugriff via DOI 
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900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6398-3  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Sharma  |D Mahima  |u Gene Regulation Laboratory, School of Biotechnology, Jawaharlal Nehru University, 110067, New Delhi, India  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Dixit  |D Aparna  |u Gene Regulation Laboratory, School of Biotechnology, Jawaharlal Nehru University, 110067, New Delhi, India  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/15(2015-08-01), 6277-6291  |x 0175-7598  |q 99:15<6277  |1 2015  |2 99  |o 253