Bacterial flagellin : a novel adjuvant for vaccine strategies
The papers of this thesis are not available in Munin: 1. Natasha A. Hynes, Clemens Furnes, Børge Nilsen Fredriksen, Tori Winther, Jarl Bøgwald, Atle N. Larsen, Roy A. Dalmo: 'Immune response of Atlantic salmon to recombinant flagellin', Vaccine (2011) vol. 29, no 44, pp 7678–7687. Availabl...
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ftunivtroemsoe:oai:munin.uit.no:10037/3798 2023-05-15T15:31:44+02:00 Bacterial flagellin : a novel adjuvant for vaccine strategies Hynes, Natasha 2011-10-18 https://hdl.handle.net/10037/3798 eng eng University of Tromsø Universitetet i Tromsø 978-82-8266-018-1 https://hdl.handle.net/10037/3798 URN:NBN:no-uit_munin_3509 openAccess Copyright 2011 The Author(s) VDP::Teknologi: 500::Bioteknologi: 590 DOKTOR-002 Doctoral thesis Doktorgradsavhandling 2011 ftunivtroemsoe 2021-06-25T17:53:11Z The papers of this thesis are not available in Munin: 1. Natasha A. Hynes, Clemens Furnes, Børge Nilsen Fredriksen, Tori Winther, Jarl Bøgwald, Atle N. Larsen, Roy A. Dalmo: 'Immune response of Atlantic salmon to recombinant flagellin', Vaccine (2011) vol. 29, no 44, pp 7678–7687. Available at http://dx.doi.org/10.1016/j.vaccine.2011.07.138 2. Natasha A. Hynes, Tiago S. Hori, Roy A. Dalmo and Matthew L. Rise: 'Functional genomics analysis of the immunological responses of Atlantic salmon (Salmo salar) spleen to injection with recombinant flagellin from Vibrio anguillarum' (manuscript) Flagellin is the principal structural protein in the locomotive organ flagellum present on flagellated bacteria, and is known to be an important evolutionarily conserved PAMP. Flagellin has been shown to bind to the PRR TLR5 which induces innate immune system responses and signaling pathways. In mammals, much focus has been placed on this protein in vaccine studies for its possible function as an adjuvant. Flagellin (FlaD from Vibrio anguillarum) was recombinantly produced in two forms, full-length (FDL) and a truncated form (FDS) with portions of the N- and C-termini removed to prevent polymerization. Both forms of flagellin were tested alone and in combination with an antigen in a dose response study to determine the most effective dosage to produce a strong immune response. A polyclonal antibody for FDS was produced and showed good specificity in immunoblot testing. Cell culture was used to compare the NF-κB response after stimulation with FDL versus FDS and resulted in a significantly larger response in cells stimulated with FDL. QPCR mRNA gene expression results showed a strong innate immune response with a number of genes known to be induced downstream by the TLR signaling pathway up- regulated including important TLR5S and inflammatory cytokines and chemokines (TNFα, IL-6, IL-8, IL-1β). Microarray analysis of salmon injected with flagellin caused reproducible gene up-regulation of inflammatory cytokines, chemokines and receptors (IL-8, TNFRSF11B, IL-1R), antimicrobial peptides (hepcidin, cathelicidin), immune genes (C/EBP, thioredoxin, C-type lectin receptor B), complement genes (complement component C7 and C7-1), peptidases (MMP-9) and genes involved in the Ras/MAPK pathway (Ras-related proteins). Down-regulation of interesting immune genes also occurred including myxovirus resistance 1, clusterin-1 and LPS-induced TNF-α homolog. Some genes exhibited early, delayed or extended response over the sampling time and flagellin seemed to be the key component to eliciting a response. Flagellin was shown to induce innate immune responses in Atlantic salmon. Doctoral or Postdoctoral Thesis Atlantic salmon Salmo salar University of Tromsø: Munin Open Research Archive Dalmo ENVELOPE(12.223,12.223,65.137,65.137) Fredriksen ENVELOPE(-45.000,-45.000,-60.733,-60.733) Furnes ENVELOPE(15.674,15.674,67.214,67.214) |
institution |
Open Polar |
collection |
University of Tromsø: Munin Open Research Archive |
op_collection_id |
ftunivtroemsoe |
language |
English |
topic |
VDP::Teknologi: 500::Bioteknologi: 590 DOKTOR-002 |
spellingShingle |
VDP::Teknologi: 500::Bioteknologi: 590 DOKTOR-002 Hynes, Natasha Bacterial flagellin : a novel adjuvant for vaccine strategies |
topic_facet |
VDP::Teknologi: 500::Bioteknologi: 590 DOKTOR-002 |
description |
The papers of this thesis are not available in Munin: 1. Natasha A. Hynes, Clemens Furnes, Børge Nilsen Fredriksen, Tori Winther, Jarl Bøgwald, Atle N. Larsen, Roy A. Dalmo: 'Immune response of Atlantic salmon to recombinant flagellin', Vaccine (2011) vol. 29, no 44, pp 7678–7687. Available at http://dx.doi.org/10.1016/j.vaccine.2011.07.138 2. Natasha A. Hynes, Tiago S. Hori, Roy A. Dalmo and Matthew L. Rise: 'Functional genomics analysis of the immunological responses of Atlantic salmon (Salmo salar) spleen to injection with recombinant flagellin from Vibrio anguillarum' (manuscript) Flagellin is the principal structural protein in the locomotive organ flagellum present on flagellated bacteria, and is known to be an important evolutionarily conserved PAMP. Flagellin has been shown to bind to the PRR TLR5 which induces innate immune system responses and signaling pathways. In mammals, much focus has been placed on this protein in vaccine studies for its possible function as an adjuvant. Flagellin (FlaD from Vibrio anguillarum) was recombinantly produced in two forms, full-length (FDL) and a truncated form (FDS) with portions of the N- and C-termini removed to prevent polymerization. Both forms of flagellin were tested alone and in combination with an antigen in a dose response study to determine the most effective dosage to produce a strong immune response. A polyclonal antibody for FDS was produced and showed good specificity in immunoblot testing. Cell culture was used to compare the NF-κB response after stimulation with FDL versus FDS and resulted in a significantly larger response in cells stimulated with FDL. QPCR mRNA gene expression results showed a strong innate immune response with a number of genes known to be induced downstream by the TLR signaling pathway up- regulated including important TLR5S and inflammatory cytokines and chemokines (TNFα, IL-6, IL-8, IL-1β). Microarray analysis of salmon injected with flagellin caused reproducible gene up-regulation of inflammatory cytokines, chemokines and receptors (IL-8, TNFRSF11B, IL-1R), antimicrobial peptides (hepcidin, cathelicidin), immune genes (C/EBP, thioredoxin, C-type lectin receptor B), complement genes (complement component C7 and C7-1), peptidases (MMP-9) and genes involved in the Ras/MAPK pathway (Ras-related proteins). Down-regulation of interesting immune genes also occurred including myxovirus resistance 1, clusterin-1 and LPS-induced TNF-α homolog. Some genes exhibited early, delayed or extended response over the sampling time and flagellin seemed to be the key component to eliciting a response. Flagellin was shown to induce innate immune responses in Atlantic salmon. |
format |
Doctoral or Postdoctoral Thesis |
author |
Hynes, Natasha |
author_facet |
Hynes, Natasha |
author_sort |
Hynes, Natasha |
title |
Bacterial flagellin : a novel adjuvant for vaccine strategies |
title_short |
Bacterial flagellin : a novel adjuvant for vaccine strategies |
title_full |
Bacterial flagellin : a novel adjuvant for vaccine strategies |
title_fullStr |
Bacterial flagellin : a novel adjuvant for vaccine strategies |
title_full_unstemmed |
Bacterial flagellin : a novel adjuvant for vaccine strategies |
title_sort |
bacterial flagellin : a novel adjuvant for vaccine strategies |
publisher |
University of Tromsø |
publishDate |
2011 |
url |
https://hdl.handle.net/10037/3798 |
long_lat |
ENVELOPE(12.223,12.223,65.137,65.137) ENVELOPE(-45.000,-45.000,-60.733,-60.733) ENVELOPE(15.674,15.674,67.214,67.214) |
geographic |
Dalmo Fredriksen Furnes |
geographic_facet |
Dalmo Fredriksen Furnes |
genre |
Atlantic salmon Salmo salar |
genre_facet |
Atlantic salmon Salmo salar |
op_relation |
978-82-8266-018-1 https://hdl.handle.net/10037/3798 URN:NBN:no-uit_munin_3509 |
op_rights |
openAccess Copyright 2011 The Author(s) |
_version_ |
1766362250704060416 |