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Increased interleukin-17 production via a phosphoinositide 3-kinase/Akt and nuclear factor kappaB-dependent pathway in patients with rheumatoid arthritis.

Kim KW, Cho ML, Park MK, Yoon CH, Park SH, Lee SH, Kim HY - Arthritis Res. Ther. (2004)

Bottom Line: Anti-CD3 antibody activated the PI3K/Akt pathway; activation of this pathway resulted in a pronounced augmentation of nuclear factor kappaB (NF-kappaB) DNA-binding activity.However, inhibition of activator protein-1 and extracellular signal-regulated kinase 1/2 did not affect IL-17 production.These results suggest that signal transduction pathways dependent on PI3K/Akt and NF-kappaB are involved in the overproduction of the key inflammatory cytokine IL-17 in RA.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Medicine, Division of Rheumatology, The Center for Rheumatic Diseases, and The Rheumatism Research Center (RhRC), Catholic Research Institutes of Medical Sciences, Catholic University of Korea, Seoul, Korea. woon1212@catholic.ac.kr

ABSTRACT
Inflammatory mediators have been recognized as being important in the pathogenesis of rheumatoid arthritis (RA). Interleukin (IL)-17 is an important regulator of immune and inflammatory responses, including the induction of proinflammatory cytokines and osteoclastic bone resorption. Evidence for the expression and proinflammatory activity of IL-17 has been demonstrated in RA synovium and in animal models of RA. Although some cytokines (IL-15 and IL-23) have been reported to regulate IL-17 production, the intracellular signaling pathways that regulate IL-17 production remain unknown. In the present study, we investigated the role of the phosphoinositide 3-kinase (PI3K)/Akt pathway in the regulation of IL-17 production in RA. Peripheral blood mononuclear cells (PBMC) from patients with RA (n = 24) were separated, then stimulated with various agents including anti-CD3, anti-CD28, phytohemagglutinin (PHA) and several inflammatory cytokines and chemokines. IL-17 levels were determined by sandwich enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction. The production of IL-17 was significantly increased in cells treated with anti-CD3 antibody with or without anti-CD28 and PHA (P < 0.05). Among tested cytokines and chemokines, IL-15, monocyte chemoattractant protein-1 and IL-6 upregulated IL-17 production (P < 0.05), whereas tumor necrosis factor-alpha, IL-1beta, IL-18 or transforming growth factor-beta did not. IL-17 was also detected in the PBMC of patients with osteoarthritis, but their expression levels were much lower than those of RA PBMC. Anti-CD3 antibody activated the PI3K/Akt pathway; activation of this pathway resulted in a pronounced augmentation of nuclear factor kappaB (NF-kappaB) DNA-binding activity. IL-17 production by activated RA PBMC is completely or partly blocked in the presence of the NF-kappaB inhibitor pyrrolidine dithiocarbamate and the PI3K/Akt inhibitor wortmannin and LY294002, respectively. However, inhibition of activator protein-1 and extracellular signal-regulated kinase 1/2 did not affect IL-17 production. These results suggest that signal transduction pathways dependent on PI3K/Akt and NF-kappaB are involved in the overproduction of the key inflammatory cytokine IL-17 in RA.

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Effects of protein kinase inhibitors and anti-rheumatic drug on anti-CD3 triggered interleukin (IL)-17 production by peripheral blood mononuclear cells (PBMC) from patients with rheumatoid arthritis. PBMC pretreated for 1 hour with pyrrolidine dithiocarbamate (PDTC; 300 μM), curcumin (10 μM), LY294002 (20 μM), wortmannin (200 nM), Cyclosporin A (500 ng/ml), dexamethasone (DEX; 100 nM), FK506 (100 ng/ml), rapamycin (10 ng/ml), SB203580 (10 nM) or PD98059 (20 μM) in combination with anti-CD3 antibody (5 μg/ml). Culture supernatant was assayed for IL-17 as described in the Materials and methods section. Each value represents the mean and SEM of three independent experiments. *, P < 0.05; **, P < 0.005.
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Figure 3: Effects of protein kinase inhibitors and anti-rheumatic drug on anti-CD3 triggered interleukin (IL)-17 production by peripheral blood mononuclear cells (PBMC) from patients with rheumatoid arthritis. PBMC pretreated for 1 hour with pyrrolidine dithiocarbamate (PDTC; 300 μM), curcumin (10 μM), LY294002 (20 μM), wortmannin (200 nM), Cyclosporin A (500 ng/ml), dexamethasone (DEX; 100 nM), FK506 (100 ng/ml), rapamycin (10 ng/ml), SB203580 (10 nM) or PD98059 (20 μM) in combination with anti-CD3 antibody (5 μg/ml). Culture supernatant was assayed for IL-17 as described in the Materials and methods section. Each value represents the mean and SEM of three independent experiments. *, P < 0.05; **, P < 0.005.

Mentions: Having observed the increased IL-17 production in RA PBMC, it was important to know which signal transduction pathways were involved. As illustrated in Fig. 3, an significant decrease in anti-CD3-induced IL-17 production was observed when co-incubated with NF-κB inhibitor, PDTC and dexamethasone in comparison with anti-CD3 alone (38 ± 5 and 54 ± 11 versus 98 ± 19 pg/ml, respectively; P < 0.05).


Increased interleukin-17 production via a phosphoinositide 3-kinase/Akt and nuclear factor kappaB-dependent pathway in patients with rheumatoid arthritis.

Kim KW, Cho ML, Park MK, Yoon CH, Park SH, Lee SH, Kim HY - Arthritis Res. Ther. (2004)

Effects of protein kinase inhibitors and anti-rheumatic drug on anti-CD3 triggered interleukin (IL)-17 production by peripheral blood mononuclear cells (PBMC) from patients with rheumatoid arthritis. PBMC pretreated for 1 hour with pyrrolidine dithiocarbamate (PDTC; 300 μM), curcumin (10 μM), LY294002 (20 μM), wortmannin (200 nM), Cyclosporin A (500 ng/ml), dexamethasone (DEX; 100 nM), FK506 (100 ng/ml), rapamycin (10 ng/ml), SB203580 (10 nM) or PD98059 (20 μM) in combination with anti-CD3 antibody (5 μg/ml). Culture supernatant was assayed for IL-17 as described in the Materials and methods section. Each value represents the mean and SEM of three independent experiments. *, P < 0.05; **, P < 0.005.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC1064895&req=5

Figure 3: Effects of protein kinase inhibitors and anti-rheumatic drug on anti-CD3 triggered interleukin (IL)-17 production by peripheral blood mononuclear cells (PBMC) from patients with rheumatoid arthritis. PBMC pretreated for 1 hour with pyrrolidine dithiocarbamate (PDTC; 300 μM), curcumin (10 μM), LY294002 (20 μM), wortmannin (200 nM), Cyclosporin A (500 ng/ml), dexamethasone (DEX; 100 nM), FK506 (100 ng/ml), rapamycin (10 ng/ml), SB203580 (10 nM) or PD98059 (20 μM) in combination with anti-CD3 antibody (5 μg/ml). Culture supernatant was assayed for IL-17 as described in the Materials and methods section. Each value represents the mean and SEM of three independent experiments. *, P < 0.05; **, P < 0.005.
Mentions: Having observed the increased IL-17 production in RA PBMC, it was important to know which signal transduction pathways were involved. As illustrated in Fig. 3, an significant decrease in anti-CD3-induced IL-17 production was observed when co-incubated with NF-κB inhibitor, PDTC and dexamethasone in comparison with anti-CD3 alone (38 ± 5 and 54 ± 11 versus 98 ± 19 pg/ml, respectively; P < 0.05).

Bottom Line: Anti-CD3 antibody activated the PI3K/Akt pathway; activation of this pathway resulted in a pronounced augmentation of nuclear factor kappaB (NF-kappaB) DNA-binding activity.However, inhibition of activator protein-1 and extracellular signal-regulated kinase 1/2 did not affect IL-17 production.These results suggest that signal transduction pathways dependent on PI3K/Akt and NF-kappaB are involved in the overproduction of the key inflammatory cytokine IL-17 in RA.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Medicine, Division of Rheumatology, The Center for Rheumatic Diseases, and The Rheumatism Research Center (RhRC), Catholic Research Institutes of Medical Sciences, Catholic University of Korea, Seoul, Korea. woon1212@catholic.ac.kr

ABSTRACT
Inflammatory mediators have been recognized as being important in the pathogenesis of rheumatoid arthritis (RA). Interleukin (IL)-17 is an important regulator of immune and inflammatory responses, including the induction of proinflammatory cytokines and osteoclastic bone resorption. Evidence for the expression and proinflammatory activity of IL-17 has been demonstrated in RA synovium and in animal models of RA. Although some cytokines (IL-15 and IL-23) have been reported to regulate IL-17 production, the intracellular signaling pathways that regulate IL-17 production remain unknown. In the present study, we investigated the role of the phosphoinositide 3-kinase (PI3K)/Akt pathway in the regulation of IL-17 production in RA. Peripheral blood mononuclear cells (PBMC) from patients with RA (n = 24) were separated, then stimulated with various agents including anti-CD3, anti-CD28, phytohemagglutinin (PHA) and several inflammatory cytokines and chemokines. IL-17 levels were determined by sandwich enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction. The production of IL-17 was significantly increased in cells treated with anti-CD3 antibody with or without anti-CD28 and PHA (P < 0.05). Among tested cytokines and chemokines, IL-15, monocyte chemoattractant protein-1 and IL-6 upregulated IL-17 production (P < 0.05), whereas tumor necrosis factor-alpha, IL-1beta, IL-18 or transforming growth factor-beta did not. IL-17 was also detected in the PBMC of patients with osteoarthritis, but their expression levels were much lower than those of RA PBMC. Anti-CD3 antibody activated the PI3K/Akt pathway; activation of this pathway resulted in a pronounced augmentation of nuclear factor kappaB (NF-kappaB) DNA-binding activity. IL-17 production by activated RA PBMC is completely or partly blocked in the presence of the NF-kappaB inhibitor pyrrolidine dithiocarbamate and the PI3K/Akt inhibitor wortmannin and LY294002, respectively. However, inhibition of activator protein-1 and extracellular signal-regulated kinase 1/2 did not affect IL-17 production. These results suggest that signal transduction pathways dependent on PI3K/Akt and NF-kappaB are involved in the overproduction of the key inflammatory cytokine IL-17 in RA.

Show MeSH
Related in: MedlinePlus