Reduction of concanavalin A-induced expression of interferon-gamma by bovine lactoferrin in feline peripheral blood mononuclear cells.
Lactoferrin (LF), a glycoprotein present in milk, mucosal secretions and neutrophils, contributes to host defense and immunomodulation. In the present study, we investigated the effect of bovine LF (bLF) on cytokine messenger RNA (mRNA) expression in concanavalin A (ConA)-stimulated feline peripheral blood mononuclear cells (PBMC). Semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and real-time PCR showed a ConA-induced increase of interferon-gamma (IFN-gamma) mRNA expression but not of interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and IL-12 p40 mRNA in feline PBMC. This ConA-induced increase of IFN-gamma mRNA expression was inhibited by addition of bLF not only 30 min before ConA stimulation but also 10, 20 and 40 min after ConA stimulation. Western blotting showed that protein tyrosine kinase (PTK) and extracellular signal-regulated kinase (ERK) in feline PBMC were activated within 10 min after the ConA stimulation and that the activation of both kinases had almost disappeared by 40 min after stimulation. Moreover, the ConA-induced IFN-gamma mRNA expression was partly prevented by genistein, a global PTK inhibitor, and PD-98059, an ERK inhibitor, respectively. These results suggest that bLF is able to inhibit the ConA-induced IFN-gamma mRNA expression by abrogation of intracellular signaling activated after interaction between ConA and its receptor.
Kobayashi
Saori
S
Department of Veterinary Clinical Science, The United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan.
Sato
Reeko
R
Inanami
Osamu
O
Yamamori
Tohru
T
Yamato
Osamu
O
Maede
Yoshimitsu
Y
Sato
Jun
J
Kuwabara
Mikinori
M
Naito
Yoshihisa
Y
D000818:Animals; D015153:Blotting, Western / Q000662:veterinary; D002415:Cats / Q000276:immunology*; D003208:Concanavalin A / Q000037:antagonists & inhibitors* / Q000276:immunology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000276:immunology; D005260:Female; D005419:Flavonoids / Q000494:pharmacology; D019833:Genistein / Q000494:pharmacology; D007371:Interferon-gamma / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D007375:Interleukin-1 / Q000235:genetics / Q000276:immunology; D018664:Interleukin-12 / Q000235:genetics / Q000276:immunology; D053762:Interleukin-12 Subunit p40; D007781:Lactoferrin / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology; D008297:Male; D021122:Protein Subunits / Q000235:genetics / Q000276:immunology; D011505:Protein-Tyrosine Kinases / Q000276:immunology; D012313:RNA / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000276:immunology
D004791:Enzyme Inhibitors; D005419:Flavonoids; D007375:Interleukin-1; D053762:Interleukin-12 Subunit p40; D021122:Protein Subunits; D014409:Tumor Necrosis Factor-alpha; D003208:Concanavalin A; D018664:Interleukin-12; D012313:RNA; D007371:Interferon-gamma; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D048049:Extracellular Signal-Regulated MAP Kinases; D007781:Lactoferrin; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one