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Fibroblasts express immune relevant genes and are important sentinel cells during tissue damage in rainbow trout (Oncorhynchus mykiss).

Ingerslev HC, Ossum CG, Lindenstrøm T, Nielsen ME - PLoS ONE (2010)

Bottom Line: Supernatant stimuli significantly increased the expression of IL-1beta, TLR-3 and TLR-9, whereas the debris stimuli only increased expression of IL-1beta.A weaker, but significant response was also seen for TLR-9 and TLR-22.Rainbow trout fibroblasts were found to be highly immune competent with a significant ability to express cytokines and immune receptors.

View Article: PubMed Central - PubMed

Affiliation: Section for Aquatic Protein Biochemistry, Division for Seafood Research, DTU Food, National Food Institute, Lyngby, Denmark. hci@aqua.dtu.dk

ABSTRACT
Fibroblasts have shown to be an immune competent cell type in mammals. However, little is known about the immunological functions of this cell-type in lower vertebrates. A rainbow trout hypodermal fibroblast cell-line (RTHDF) was shown to be responsive to PAMPs and DAMPs after stimulation with LPS from E. coli, supernatant and debris from sonicated RTHDF cells. LPS was overall the strongest inducer of IL-1beta, IL-8, IL-10, TLR-3 and TLR-9. IL-1beta and IL-8 were already highly up regulated after 1 hour of LPS stimulation. Supernatant stimuli significantly increased the expression of IL-1beta, TLR-3 and TLR-9, whereas the debris stimuli only increased expression of IL-1beta. Consequently, an in vivo experiment was further set up. By mechanically damaging the muscle tissue of rainbow trout, it was shown that fibroblasts in the muscle tissue of rainbow trout contribute to electing a highly local inflammatory response following tissue injury. The damaged muscle tissue showed a strong increase in the expression of the immune genes IL-1beta, IL-8 and TGF-beta already 4 hours post injury at the site of injury while the expression in non-damaged muscle tissue was not influenced. A weaker, but significant response was also seen for TLR-9 and TLR-22. Rainbow trout fibroblasts were found to be highly immune competent with a significant ability to express cytokines and immune receptors. Thus fish fibroblasts are believed to contribute significantly to local inflammatory reactions in concert with the traditional immune cells.

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Related in: MedlinePlus

Damage procedures and sampling from mechanically injured rainbow trout.The fish were injured on the left side behind the dorsal fin using the damage instrument. Muscle tissue samples were taken in the injured area while the internal control samples were taken from the same place relative to the dorsal fin on the right side of the fish (A). The vertical position of the site of injury/sampling site is shown in (B). Sampling of muscle tissue from non-injured control fish was performed in the same area as shown for injured fish (not shown).
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pone-0009304-g003: Damage procedures and sampling from mechanically injured rainbow trout.The fish were injured on the left side behind the dorsal fin using the damage instrument. Muscle tissue samples were taken in the injured area while the internal control samples were taken from the same place relative to the dorsal fin on the right side of the fish (A). The vertical position of the site of injury/sampling site is shown in (B). Sampling of muscle tissue from non-injured control fish was performed in the same area as shown for injured fish (not shown).

Mentions: The device used for mechanical tissue damage is shown in figure 3 A–B. The following effects of mechanical injury of the muscle tissue on the expression of the genes IL-1β, IL-8, TGF-β and TLR-3, 5 m, 9 and 22 at the time points 4, 8 and 24 hours post injury (p.i.) is shown in figure 4 A-F. No significant changes were seen for IL-10 (P>0.05) (not illustrated). The study showed a strong induction of immune related genes, especially of the cytokines IL-1β, IL-8 and TGF-β in the injured muscle tissue at all samplings. The average up-regulation in the injured fish for the three sampling points was between 5.2 to 21.8 folds for IL-1β; 20.0 to 59.3 folds for IL-8 and 4.9 to 13.9 folds for TGF- β. For all three genes the mean expression level was increasing from 4 hours to 24 hours post injury and the expression at the site of injury was significantly higher at all sampling points compared to the non-injured samples from the same fish (P<0.05).


Fibroblasts express immune relevant genes and are important sentinel cells during tissue damage in rainbow trout (Oncorhynchus mykiss).

Ingerslev HC, Ossum CG, Lindenstrøm T, Nielsen ME - PLoS ONE (2010)

Damage procedures and sampling from mechanically injured rainbow trout.The fish were injured on the left side behind the dorsal fin using the damage instrument. Muscle tissue samples were taken in the injured area while the internal control samples were taken from the same place relative to the dorsal fin on the right side of the fish (A). The vertical position of the site of injury/sampling site is shown in (B). Sampling of muscle tissue from non-injured control fish was performed in the same area as shown for injured fish (not shown).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2823790&req=5

pone-0009304-g003: Damage procedures and sampling from mechanically injured rainbow trout.The fish were injured on the left side behind the dorsal fin using the damage instrument. Muscle tissue samples were taken in the injured area while the internal control samples were taken from the same place relative to the dorsal fin on the right side of the fish (A). The vertical position of the site of injury/sampling site is shown in (B). Sampling of muscle tissue from non-injured control fish was performed in the same area as shown for injured fish (not shown).
Mentions: The device used for mechanical tissue damage is shown in figure 3 A–B. The following effects of mechanical injury of the muscle tissue on the expression of the genes IL-1β, IL-8, TGF-β and TLR-3, 5 m, 9 and 22 at the time points 4, 8 and 24 hours post injury (p.i.) is shown in figure 4 A-F. No significant changes were seen for IL-10 (P>0.05) (not illustrated). The study showed a strong induction of immune related genes, especially of the cytokines IL-1β, IL-8 and TGF-β in the injured muscle tissue at all samplings. The average up-regulation in the injured fish for the three sampling points was between 5.2 to 21.8 folds for IL-1β; 20.0 to 59.3 folds for IL-8 and 4.9 to 13.9 folds for TGF- β. For all three genes the mean expression level was increasing from 4 hours to 24 hours post injury and the expression at the site of injury was significantly higher at all sampling points compared to the non-injured samples from the same fish (P<0.05).

Bottom Line: Supernatant stimuli significantly increased the expression of IL-1beta, TLR-3 and TLR-9, whereas the debris stimuli only increased expression of IL-1beta.A weaker, but significant response was also seen for TLR-9 and TLR-22.Rainbow trout fibroblasts were found to be highly immune competent with a significant ability to express cytokines and immune receptors.

View Article: PubMed Central - PubMed

Affiliation: Section for Aquatic Protein Biochemistry, Division for Seafood Research, DTU Food, National Food Institute, Lyngby, Denmark. hci@aqua.dtu.dk

ABSTRACT
Fibroblasts have shown to be an immune competent cell type in mammals. However, little is known about the immunological functions of this cell-type in lower vertebrates. A rainbow trout hypodermal fibroblast cell-line (RTHDF) was shown to be responsive to PAMPs and DAMPs after stimulation with LPS from E. coli, supernatant and debris from sonicated RTHDF cells. LPS was overall the strongest inducer of IL-1beta, IL-8, IL-10, TLR-3 and TLR-9. IL-1beta and IL-8 were already highly up regulated after 1 hour of LPS stimulation. Supernatant stimuli significantly increased the expression of IL-1beta, TLR-3 and TLR-9, whereas the debris stimuli only increased expression of IL-1beta. Consequently, an in vivo experiment was further set up. By mechanically damaging the muscle tissue of rainbow trout, it was shown that fibroblasts in the muscle tissue of rainbow trout contribute to electing a highly local inflammatory response following tissue injury. The damaged muscle tissue showed a strong increase in the expression of the immune genes IL-1beta, IL-8 and TGF-beta already 4 hours post injury at the site of injury while the expression in non-damaged muscle tissue was not influenced. A weaker, but significant response was also seen for TLR-9 and TLR-22. Rainbow trout fibroblasts were found to be highly immune competent with a significant ability to express cytokines and immune receptors. Thus fish fibroblasts are believed to contribute significantly to local inflammatory reactions in concert with the traditional immune cells.

Show MeSH
Related in: MedlinePlus