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Upregulation of TNF-α Production by IFN-γ and LPS in Cultured Canine Keratinocytes: Application to Monosaccharides Effects

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Abstract

Activated keratinocytes play a key role in the cutaneous immune system by their interactions with other cell types through the production of cytokines with both autocrine and paracrine activity. But there is little knowledge about epidermal cytokines in the dog. In this study, cultured canine keratinocytes were stimulated by human recombinant interferon γ (IFN-γ) and lipopolysaccharide (LPS) and cell supernatants were tested for tumour necrosis factor α (TNF-α) concentration using a cell viability assay on a murine cell line. We show that IFN-γ in combination with LPS significantly increases TNF-α secretion by canine keratinocytes. The best stimulations were obtained using confluent cultures and the association of IFN-γ (400 ng/ml) and LPS (40 μg/ml). The experimental protocol we describe represents a new method for studying keratinocyte activation and its modulation in the dog. We provide an example of application of our method: the study of the effects of different monosaccharides on canine keratinocyte activation.

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Abbreviations

IFN:

interferon

LPS:

lipopolysaccharide

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

OD:

optical density

SDS:

sodium dodecyl sulphate

TNF:

tumour necrosis factor

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Authors and Affiliations

  1. Unit of Dermatology, Clinical Parasitology and Mycology, National Veterinary School, Atlanpole – La Chantrerie, BP 40 706, 44 307, Nantes Cedex 03, France

    C. Ibisch, P. Bourdeau & C. Cadiot

  2. Unité INSERM 346, Hôpital E. Herriot, Lyon Cedex, France

    J. Viac

  3. Virbac S.A. Laboratories, Carros, France

    H. Gatto

Authors
  1. C. Ibisch
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  2. P. Bourdeau
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  3. C. Cadiot
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  4. J. Viac
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  5. H. Gatto
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Correspondence to C. Ibisch.

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Ibisch, C., Bourdeau, P., Cadiot, C. et al. Upregulation of TNF-α Production by IFN-γ and LPS in Cultured Canine Keratinocytes: Application to Monosaccharides Effects. Vet Res Commun 31, 835–846 (2007). https://doi.org/10.1007/s11259-007-0004-9

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  • DOI: https://doi.org/10.1007/s11259-007-0004-9

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