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The role of IL-10 in iNOS and cytokine mRNA expression during in vitro differentiation of bovine mononuclear phagocytes.
- Source :
-
Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research [J Interferon Cytokine Res] 1998 Mar; Vol. 18 (3), pp. 139-49. - Publication Year :
- 1998
-
Abstract
- In the study reported here, we used RT-PCR with primers specific for interleukin-1 (IL-1), IL-6, IL-10, tumor necrosis factor-alpha (TNF-alpha), and nitric oxide synthase (iNOS) to assess the cytokine mRNA expression associated with bovine blood monocytes during their differentiation to macrophages cultured on plastic (1 week). In addition, we used RT-PCR to assess the contribution of gammadelta T cells as a source of interferon-gamma (IFN-gamma), the induction signal for iNOS. Further, we evaluated cytocentrifuge preparations from the cultures for the production of IL-10 using specific antibody. We previously demonstrated that iNOS can be induced in cultured bovine monocytes in response to IFN-gamma and TNF-alpha but lose this capability in a short period of time. However, we demonstrate here that iNOS induction from monocytes cultured with IFN-gamma secreting gammadelta T cells is prolonged, suggesting that this source of IFN-gamma primes the monocytes before exogenous stimulation. Based on mRNA expression, placement of monocytes in culture resulted in activation, followed by quiescence. By 6 days in culture, the iNOS message was reduced below the basal level. In addition, the TNF-alpha message was substantially reduced, and IL-1 and IL-6 messages were reduced below detectable levels. This correlated with an increase in IL-10 message. Downregulation of these same cytokine messages as well as IFN-gamma message occurred within a 20-h period when IL-10 was added exogenously to cultures of total leukocytes. At the same time, there was an increase in the number of IL-10-positive cells and an increase in the intensity of anti-IL-10 staining within adherent cells. These results provide evidence for IL-10 regulation of some bovine mononuclear phagocyte effector functions.
- Subjects :
- Animals
Cattle
Cell Adhesion
Cell Differentiation physiology
Down-Regulation
Enzyme Induction
Interferon-gamma biosynthesis
Leukocytes, Mononuclear cytology
Macrophages cytology
Nitric Oxide biosynthesis
Nitric Oxide Synthase Type II
Polymerase Chain Reaction methods
Transcription, Genetic
Cytokines genetics
Interleukin-10 physiology
Leukocytes, Mononuclear metabolism
Macrophages metabolism
Nitric Oxide Synthase genetics
RNA, Messenger biosynthesis
Subjects
Details
- Language :
- English
- ISSN :
- 1079-9907
- Volume :
- 18
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research
- Publication Type :
- Academic Journal
- Accession number :
- 9555975
- Full Text :
- https://doi.org/10.1089/jir.1998.18.139