Kinetics of ovine interferon-gamma production: detection of mRNA and characterisation of biological activity.
The kinetics of interferon-gamma (IFN-gamma) production were studied in sheep mesenteric lymph node (MLN) cells at the molecular level using an ovine IFN-gamma cDNA probe and by bioassay which was verified by blocking antiviral activity with a monoclonal antibody (Mab) against recombinant bovine IFN-gamma IFN-gamma mRNA appeared in MLN cells within 4 h of stimulation with phorbol ester and Concanavalin A and was not detectable by 72 h after stimulation. Biologically active IFN-gamma appeared in the culture supernatants 8 h after stimulation and was still present 96 h later when de novo synthesis had terminated. Acid dialysis and Mab neutralisation demonstrated conclusively that native ovine IFN-gamma is a pH 2 labile cytokine.
Entrican
G
G
Moredun Research Institute, Edinburgh, UK.
McInnes
C J
CJ
Rothel
J S
JS
Haig
D M
DM
D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D002522:Chlorocebus aethiops; D003470:Culture Media; D007371:Interferon-gamma / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D007668:Kidney; D007700:Kinetics; D008198:Lymph Nodes; D008643:Mesentery; D009500:Neutralization Tests; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D012756:Sheep
D000911:Antibodies, Monoclonal; D003470:Culture Media; D012333:RNA, Messenger; D007371:Interferon-gamma