Cytokine mRNA levels in isolated feline monocytes.
Real-time PCR systems were developed to quantitate cytokine expression in short-time cultivated feline monocytes. Feline-specific interleukin-1beta (IL-1beta), IL-6, and tumor necrosis factor-alpha (TNF-alpha) primers as well as TaqMan probes were designed and were adapted to a quantitative PCR system which had been previously established for feline IL-10 and IL-12 p40. Quantitative analysis of cytokine messenger RNA (mRNA) transcription based on the comparison of the cytokine with the housekeeping gene feline glyceraldehyde-3-phosphate dehydrogenase (GAPDH), providing universally expressed mRNA. GAPDH mRNA was readily detectable in cDNA prepared from short-time cultivated peripheral blood monocytes. Cytokine mRNA was demonstrated in all samples at variable amounts. IL-1beta and TNF-alpha mRNA was constitutively expressed whereas IL-6, IL-10 and IL-12 p40 mRNA was generally expressed at a lower level and was occasionally not detected. There was a great variability of cytokine production between individual cats and at different time points in the same cat.
Kipar
A
A
Institute for Veterinary Pathology, Frankfurter Strasse 96, Justus-Liebig University Giessen, D-35392 Giessen, Germany. anja.kipar@vetmed.uni-giessen.de
Leutenegger
C M
CM
Hetzel
U
U
Akens
M K
MK
Mislin
C N
CN
Reinacher
M
M
Lutz
H
H
D000818:Animals; D002415:Cats / Q000097:blood*; D002469:Cell Separation / Q000662:veterinary; D016207:Cytokines / Q000235:genetics*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D007375:Interleukin-1 / Q000235:genetics; D015850:Interleukin-6 / Q000235:genetics; D009000:Monocytes / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012313:RNA; D012333:RNA, Messenger / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
D016207:Cytokines; D007375:Interleukin-1; D015850:Interleukin-6; C034477:RNA primers; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D012313:RNA; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases