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Effect of Different Adjuvants on Protection and Side-Effects Induced by Helicobacter suis Whole-Cell Lysate Vaccination.

Bosschem I, Bayry J, De Bruyne E, Van Deun K, Smet A, Vercauteren G, Ducatelle R, Haesebrouck F, Flahou B - PLoS ONE (2015)

Bottom Line: Therefore, we decided to test alternative strategies, including sublingual vaccine administration, to reduce the unwanted side-effects.In the groups immunized subcutaneously with FC/lysate and CCR4 antagonist/lysate, a significant protection was observed.In general, an inverse correlation was observed between IFN-γ, IL-4, IL-17, KC, MIP-2 and LIX mRNA expression and H. suis colonization density, whereas lower IL-10 expression levels were observed in partially protected animals.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.

ABSTRACT
Helicobacter suis (H. suis) is a widespread porcine gastric pathogen, which is also of zoonotic importance. The first goal of this study was to investigate the efficacy of several vaccine adjuvants (CpG-DNA, Curdlan, Freund's Complete and Incomplete, Cholera toxin), administered either subcutaneously or intranasally along with H. suis whole-cell lysate, to protect against subsequent H. suis challenge in a BALB/c infection model. Subcutaneous immunization with Freund's complete (FC)/lysate and intranasal immunization with Cholera toxin (CT)/lysate were shown to be the best options for vaccination against H. suis, as determined by the amount of colonizing H. suis bacteria in the stomach, although adverse effects such as post-immunization gastritis/pseudo-pyloric metaplasia and increased mortality were observed, respectively. Therefore, we decided to test alternative strategies, including sublingual vaccine administration, to reduce the unwanted side-effects. A CCR4 antagonist that transiently inhibits the migration of regulatory T cells was also included as a new adjuvant in this second study. Results confirmed that immunization with CT (intranasally or sublingually) is among the most effective vaccination protocols, but increased mortality was still observed. In the groups immunized subcutaneously with FC/lysate and CCR4 antagonist/lysate, a significant protection was observed. Compared to the FC/lysate immunized group, gastric pseudo-pyloric metaplasia was less severe or even absent in the CCR4 antagonist/lysate immunized group. In general, an inverse correlation was observed between IFN-γ, IL-4, IL-17, KC, MIP-2 and LIX mRNA expression and H. suis colonization density, whereas lower IL-10 expression levels were observed in partially protected animals.

No MeSH data available.


Related in: MedlinePlus

Relative gene expression of cytokines and chemokines in the stomach of challenged animals after immunization or after sham inoculation in study 1.The first bar represents the pooled data of the animals that were sham-immunized with HBSS (intranasally and subcutaneously) and that were not challenged with H. suis (Neg. con). The second bar represents the pooled data of the animals that were sham-immunized with HBSS (intranasally and subcutaneously) and that were challenged with H. suis (Pos. con). Bars 3, 4 and 5 represent the groups of animals that were immunized subcutaneously with Freund’s complete (FC/lysate/SC), Freund’s incomplete (FIC/lysate/SC) or Curdlan (Curd/lysate/SC) and challenged with H. suis. Bars 6, 7 and 8 represent the animals that were immunized intranasally with Cholera Toxin (CT/lysate/IN), CpG-DNA (CpG/lysate/IN) or Curdlan (Curd/lysate/IN) and challenged with H. suis. An * (p<0.05) indicates a significant modulation of mRNA expression levels compared to the sham-immunized/challenged groups. Cytokine expression between immunized and challenged groups was also compared with each other. When no differences could be found between the different immunized groups, the same letter designation was attributed.
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pone.0131364.g008: Relative gene expression of cytokines and chemokines in the stomach of challenged animals after immunization or after sham inoculation in study 1.The first bar represents the pooled data of the animals that were sham-immunized with HBSS (intranasally and subcutaneously) and that were not challenged with H. suis (Neg. con). The second bar represents the pooled data of the animals that were sham-immunized with HBSS (intranasally and subcutaneously) and that were challenged with H. suis (Pos. con). Bars 3, 4 and 5 represent the groups of animals that were immunized subcutaneously with Freund’s complete (FC/lysate/SC), Freund’s incomplete (FIC/lysate/SC) or Curdlan (Curd/lysate/SC) and challenged with H. suis. Bars 6, 7 and 8 represent the animals that were immunized intranasally with Cholera Toxin (CT/lysate/IN), CpG-DNA (CpG/lysate/IN) or Curdlan (Curd/lysate/IN) and challenged with H. suis. An * (p<0.05) indicates a significant modulation of mRNA expression levels compared to the sham-immunized/challenged groups. Cytokine expression between immunized and challenged groups was also compared with each other. When no differences could be found between the different immunized groups, the same letter designation was attributed.

Mentions: In the first study, data from immunized/challenged groups were compared to pooled data from the sham-immunized/challenged positive control groups (Fig 8). Expression of IL-10, an important immunoregulatory cytokine, was significantly lower in all the adjuvant groups as compared to the sham-immunized and challenged groups (p<0.0001). In the subcutaneously immunized/challenged groups, adjuvants showed distinct differences in their ability to mount helper T cell responses. Although FC/lysate, FIC/ lysate and and Curdlan/lysate groups showed significantly higher Th1 and Th2 cytokine signatures (IFN-γ; p<0.0001 and IL-4; p<0.01), Th17 cytokine responses (IL-17) were only observed with Freund’s complete and incomplete adjuvant groups. Mice immunized by mucosal routes displayed distinct helper T cell responses as compared to subcutaneous immunization routes. No Th2 response (IL-4) was induced in CT/lysate, CpG/lysate and Curdlan/lysate immunized groups, suggesting that these adjuvants selectively induce Th1 and Th17 responses when administered by mucosal routes. Clearly, Curdlan/lysate, irrespective of the route of vaccination, was less efficient to induce chemokine expression as compared to Freund’s adjuvants. In all immunized/challenged animals, LIX showed higher mRNA expression levels (p<0.05) when compared to the sham-immunized/challenged animals. This was not the case for MIP-2 and KC. However, a clear upregulation of MIP-2 mRNA expression levels was observed in the FC/lysate and FIC/lysate groups, while a clear upregulation of KC expression was only observed in the FC/lysate and CT/lysate groups. Messenger RNA expression levels of these cytokines are presented in Fig 8. (Fig 8. Relative gene expression in the stomach in challenged animals after immunization or after sham inoculation in study 1)


Effect of Different Adjuvants on Protection and Side-Effects Induced by Helicobacter suis Whole-Cell Lysate Vaccination.

Bosschem I, Bayry J, De Bruyne E, Van Deun K, Smet A, Vercauteren G, Ducatelle R, Haesebrouck F, Flahou B - PLoS ONE (2015)

Relative gene expression of cytokines and chemokines in the stomach of challenged animals after immunization or after sham inoculation in study 1.The first bar represents the pooled data of the animals that were sham-immunized with HBSS (intranasally and subcutaneously) and that were not challenged with H. suis (Neg. con). The second bar represents the pooled data of the animals that were sham-immunized with HBSS (intranasally and subcutaneously) and that were challenged with H. suis (Pos. con). Bars 3, 4 and 5 represent the groups of animals that were immunized subcutaneously with Freund’s complete (FC/lysate/SC), Freund’s incomplete (FIC/lysate/SC) or Curdlan (Curd/lysate/SC) and challenged with H. suis. Bars 6, 7 and 8 represent the animals that were immunized intranasally with Cholera Toxin (CT/lysate/IN), CpG-DNA (CpG/lysate/IN) or Curdlan (Curd/lysate/IN) and challenged with H. suis. An * (p<0.05) indicates a significant modulation of mRNA expression levels compared to the sham-immunized/challenged groups. Cytokine expression between immunized and challenged groups was also compared with each other. When no differences could be found between the different immunized groups, the same letter designation was attributed.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4482594&req=5

pone.0131364.g008: Relative gene expression of cytokines and chemokines in the stomach of challenged animals after immunization or after sham inoculation in study 1.The first bar represents the pooled data of the animals that were sham-immunized with HBSS (intranasally and subcutaneously) and that were not challenged with H. suis (Neg. con). The second bar represents the pooled data of the animals that were sham-immunized with HBSS (intranasally and subcutaneously) and that were challenged with H. suis (Pos. con). Bars 3, 4 and 5 represent the groups of animals that were immunized subcutaneously with Freund’s complete (FC/lysate/SC), Freund’s incomplete (FIC/lysate/SC) or Curdlan (Curd/lysate/SC) and challenged with H. suis. Bars 6, 7 and 8 represent the animals that were immunized intranasally with Cholera Toxin (CT/lysate/IN), CpG-DNA (CpG/lysate/IN) or Curdlan (Curd/lysate/IN) and challenged with H. suis. An * (p<0.05) indicates a significant modulation of mRNA expression levels compared to the sham-immunized/challenged groups. Cytokine expression between immunized and challenged groups was also compared with each other. When no differences could be found between the different immunized groups, the same letter designation was attributed.
Mentions: In the first study, data from immunized/challenged groups were compared to pooled data from the sham-immunized/challenged positive control groups (Fig 8). Expression of IL-10, an important immunoregulatory cytokine, was significantly lower in all the adjuvant groups as compared to the sham-immunized and challenged groups (p<0.0001). In the subcutaneously immunized/challenged groups, adjuvants showed distinct differences in their ability to mount helper T cell responses. Although FC/lysate, FIC/ lysate and and Curdlan/lysate groups showed significantly higher Th1 and Th2 cytokine signatures (IFN-γ; p<0.0001 and IL-4; p<0.01), Th17 cytokine responses (IL-17) were only observed with Freund’s complete and incomplete adjuvant groups. Mice immunized by mucosal routes displayed distinct helper T cell responses as compared to subcutaneous immunization routes. No Th2 response (IL-4) was induced in CT/lysate, CpG/lysate and Curdlan/lysate immunized groups, suggesting that these adjuvants selectively induce Th1 and Th17 responses when administered by mucosal routes. Clearly, Curdlan/lysate, irrespective of the route of vaccination, was less efficient to induce chemokine expression as compared to Freund’s adjuvants. In all immunized/challenged animals, LIX showed higher mRNA expression levels (p<0.05) when compared to the sham-immunized/challenged animals. This was not the case for MIP-2 and KC. However, a clear upregulation of MIP-2 mRNA expression levels was observed in the FC/lysate and FIC/lysate groups, while a clear upregulation of KC expression was only observed in the FC/lysate and CT/lysate groups. Messenger RNA expression levels of these cytokines are presented in Fig 8. (Fig 8. Relative gene expression in the stomach in challenged animals after immunization or after sham inoculation in study 1)

Bottom Line: Therefore, we decided to test alternative strategies, including sublingual vaccine administration, to reduce the unwanted side-effects.In the groups immunized subcutaneously with FC/lysate and CCR4 antagonist/lysate, a significant protection was observed.In general, an inverse correlation was observed between IFN-γ, IL-4, IL-17, KC, MIP-2 and LIX mRNA expression and H. suis colonization density, whereas lower IL-10 expression levels were observed in partially protected animals.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.

ABSTRACT
Helicobacter suis (H. suis) is a widespread porcine gastric pathogen, which is also of zoonotic importance. The first goal of this study was to investigate the efficacy of several vaccine adjuvants (CpG-DNA, Curdlan, Freund's Complete and Incomplete, Cholera toxin), administered either subcutaneously or intranasally along with H. suis whole-cell lysate, to protect against subsequent H. suis challenge in a BALB/c infection model. Subcutaneous immunization with Freund's complete (FC)/lysate and intranasal immunization with Cholera toxin (CT)/lysate were shown to be the best options for vaccination against H. suis, as determined by the amount of colonizing H. suis bacteria in the stomach, although adverse effects such as post-immunization gastritis/pseudo-pyloric metaplasia and increased mortality were observed, respectively. Therefore, we decided to test alternative strategies, including sublingual vaccine administration, to reduce the unwanted side-effects. A CCR4 antagonist that transiently inhibits the migration of regulatory T cells was also included as a new adjuvant in this second study. Results confirmed that immunization with CT (intranasally or sublingually) is among the most effective vaccination protocols, but increased mortality was still observed. In the groups immunized subcutaneously with FC/lysate and CCR4 antagonist/lysate, a significant protection was observed. Compared to the FC/lysate immunized group, gastric pseudo-pyloric metaplasia was less severe or even absent in the CCR4 antagonist/lysate immunized group. In general, an inverse correlation was observed between IFN-γ, IL-4, IL-17, KC, MIP-2 and LIX mRNA expression and H. suis colonization density, whereas lower IL-10 expression levels were observed in partially protected animals.

No MeSH data available.


Related in: MedlinePlus