Quantitative real-time RT-PCR measurement of cytokine mRNA expression in the skin of normal cats and cats with allergic skin disease

Veterinary Immunology and Immunopathology
K TaglingerA P Foster

Abstract

Feline allergic skin disease is thought to be associated with dermal infiltration of Th(2) lymphocytes and synthesis of associated cytokines. In this study, real-time RT-PCR assays were developed to measure feline interleukin (IL)-2, IL-4, IL-5, IL-6, IL-10, IL12 (p35 and p40), IL-18, tumour necrosis factor-alpha (TNF-alpha), transforming growth factor-beta (TGF-beta), interferon-gamma (IFN-gamma) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA in the skin of healthy control cats, and in the lesional and non-lesional skin of cats with allergic skin disease. Total RNA was extracted from skin biopsies using the RNeasy Mini Kit with on-column and in-solution DNase digestion steps. cDNA was synthesised using Improm-II reverse transcriptase and random hexamers. Real-time PCR was carried out using an iCycler IQ system (Bio-Rad), and gene-specific primers were designed to span an exon/exon junction of each cytokine gene. Taq-man probes were used to add specificity to the system. Messenger RNA from the housekeeping gene GAPDH was used for normalisation of the cytokine threshold cycle. The eleven cytokine mRNA transcripts quantified were present at varying levels, but there was no apparent difference in expression between norm...Continue Reading

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Citations

Mar 19, 2014·Veterinary Immunology and Immunopathology·Shanna L SealsCherie M Pucheu-Haston
May 28, 2013·In Vitro Cellular & Developmental Biology. Animal·Mahdieh Sadat TaghaviGholamreza Javadi
Aug 5, 2015·Cell and Tissue Research·Grace AmpemTamás Röszer
Jul 2, 2011·Veterinary Immunology and Immunopathology·Céline Robert-TissotHans Lutz
Oct 13, 2009·Veterinary Immunology and Immunopathology·Carol R Reinero
Jun 27, 2009·Veterinary Immunology and Immunopathology·Henriëtte E Booij-VrielingHerman A W Hazewinkel
Nov 21, 2014·Clinical and Vaccine Immunology : CVI·Chelsea A E HicksSéverine Tasker
Mar 14, 2009·Environmental Research·Carol R ReineroPeter Rabinowitz
Jul 19, 2016·Veterinary Dermatology·Verena K Affolter
Jan 21, 2021·Veterinary Dermatology·Richard HalliwellThierry Olivry
Jun 29, 2021·Veterinary Dermatology·Cheryl VargoFrane Banovic
Sep 15, 2021·Veterinary Dermatology·Caitlin E OlderAline Rodrigues Hoffmann

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