title: Cloning of fox (Vulpes vulpes) Il2, Il6, Il10 and IFNgamma and analysis of their expression by quantitative RT-PCR in fox PBMC after in vitro stimulation by Concanavalin A.
abstract: The immune response in the fox (Vulpes vulpes), despite the success of the oral rabies vaccine is not well characterised, and specific immunological tools are needed. A quantitative RT-PCR using SyBR Green to investigate fox cytokine expression after antigen PBMC in vitro re-stimulation is presented here. First, we cloned by homology with dog cytokine sequences the fox IL2, IL6, IL10, IFNgamma and a partial 18S sequence. Fox specific primers were then defined and used to set up a species-specific quantitative RT-PCR assay using SyBR Green and 18S housekeeping gene as internal standard. The technique was validated using total RNA from fox PBMC stimulated with a polyclonal activator, Concanavaline A.
authors.affiliation: AFSSA Nancy, Laboratoire d'Etudes et de Recherches sur la Rage et la Pathologie des Animaux Sauvages, B.P. 9, 54220 Malzéville, France. m.rolland@afssa.fr
authors.forename: Magali
authors.initials: M
authors.lastname: Rolland-Turner
authors.forename: Guillaume
authors.initials: G
authors.lastname: Farré
authors.forename: Franck
authors.initials: F
authors.lastname: Boué
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002415:Cats; D003001:Cloning, Molecular; D003208:Concanavalin A / Q000494:pharmacology*; D004285:Dogs; D005589:Foxes / Q000235:genetics*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D007371:Interferon-gamma / Q000737:chemistry / Q000235:genetics*; D016753:Interleukin-10 / Q000737:chemistry / Q000235:genetics*; D007376:Interleukin-2 / Q000737:chemistry / Q000235:genetics*; D015850:Interleukin-6 / Q000737:chemistry / Q000235:genetics*; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013997:Time Factors
chemical_list: D007376:Interleukin-2; D015850:Interleukin-6; D003208:Concanavalin A; D016753:Interleukin-10; D007371:Interferon-gamma