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. 2010 Nov;45(11):834-41.
doi: 10.1016/j.exger.2010.06.007. Epub 2010 Jul 1.

Prostaglandin E2-dependent IL-23 production in aged murine dendritic cells

Rebecca G Myer  1 , Rabab El Mezayen, Kevin P High
Affiliations

Prostaglandin E2-dependent IL-23 production in aged murine dendritic cells

Rebecca G Myer et al. Exp Gerontol. 2010 Nov.

Abstract

CD4+ T cells of the Th17 subtype are over-represented in the aged immune system. Dendritic cells (DC) play a critical role in naïve CD4+ T cell differentiation. However, expression of cytokines by aged DC that promote differentiation or survival of Th17 cells has not been extensively investigated. Using bone marrow-derived DC from C57BL/6 mice of different ages we compared cytokine production after DC activation by Toll-like receptor agonists for TLR4 and/or TLR7/8. DC-derived TNF-α and IL-12p70 production and expression of DC co-stimulatory molecules did not vary significantly by age indicating that TLR expression, function and signal transduction were intact in aged DC. There were relatively minor age-related changes in TGF-β and IL-6 which promote Th17 differentiation, but IL-23, a Th17-suvival cytokine, increased more than 40-fold across the lifespan. DC-derived prostaglandin E2 (PGE2) also increased with age and the up-regulation of IL-23 expression by aged DC was blocked by indomethacin that prevents PGE2 production, and by antagonists of PGE2 receptors. Exogenous PGE2 added to DC cultures further enhanced IL-23 production from aged but not young DCs. These data indicate that age-related changes in DC PGE2 production are necessary, but not sufficient to induce DC IL-23 production. Such changes may play a role in the expansion of Th17 cells in the aged immune system.

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Figures

Figure 1
Figure 1. Age-related changes in DCs expression of TNF-α, IL-12 and IL-10
Bone marrow progenitor cells were isolated from C57BL/6 mice (n = 4–5 mice), differentiated with GM-CSF for six days, and then stimulated with the indicated TLR agonist, individually or in combination. After 12 h, culture supernatants were collected, and cytokines levels were measured by ELISA. Samples were assayed in triplicate. Data represent the mean ± SEM from three experiments. *, statistically significant p≤0.05 (18.months vs. 2, 6, or 12 months).
Figure 2
Figure 2. Expression of CD80 and CD86 on DCs from young and old mice after TLR stimulation
Bone marrow progenitor cells from 2 and 18 months old C57BL/6 mice were differentiated with GM-CSF for six days, and then stimulated with the indicated TLR agonist, individually or in combination. After 24 hours (A) CD80 or (B) CD86 was measured by flow cytometry after labeling with fluorescent antibodies. These are representative experiments. (CD80 n= 3, CD86 n= 4).
Figure 3
Figure 3. Expression of Th17-favorable mediators (Compounds) IL-23, PGE2 and TGF-β is significantly increased in aged DCs upon TLR activation
Bone marrow progenitor cells were isolated from C57BL/6 mice (n = 4–5 mice), differentiated with GM-CSF for six days, and then stimulated with the indicated TLR agonist, individually or in combination. After 9 h, total RNA was isolated and analyzed for gene expression by real-time PCR. Culture supernatants were collected after 12 hours, and cytokines levels were measured by ELISA. Samples were assayed in triplicate. Data represent the mean ± SEM from three experiments. *, statistically significant p≤0.05 (18 months vs. 2, 6, or 12 months).
Figure 4
Figure 4. PGE2 upregulates IL-23 production from aged DCs
(A) PGE2 increases IL-23 production by aged DCs. Bone marrow progenitor cells were isolated from C57BL/6 mice (n = 5 mice), differentiated with GM-CSF for six days then cultured in media with or without 10 mM PGE2. After 1h, LPS+R848 was added for an additional 6 h. IL-23 levels in the culture supernatants were determined by ELISA. (B) Indomethacin decreases IL-23 production that is induced by LPS+R848 in aged DCs. DCs were isolated and grown with GMCSF for 6 days then treated with 10 μM Indomethacin, After 24 h, LPS+R848 was added for an additional 6 h. IL-23 levels in the culture supernatants were determined by ELISA. (C) EP2 and/or EP4 antagonist reduces IL-23 induction by LPS+R848. DCs were isolated and stimulated as above then treated with 50μM of EP2 antagonist (AH6809), EP4 antagonist (AH 23848), or EP2+EP4 antagonists. After 1 h, LPS+R848 was added for an additional 6 h, IL-23 levels in the culture supernatants were determined by ELISA. Samples were assayed in triplicate. Data represent the mean ± SEM from three experiments. *, statistically significant (p<0.05), 18 months vs. 2 months.
Figure 5
Figure 5. Aging does not influence EP2 or EP4 expression in DCs
Bone marrow progenitor cells were isolated from C57BL/6 mice (n = 5 mice), differentiated with GM-CSF for six days were left unstimulated or stimulated with LPS+R848 for 6 hours. Total RNA was isolated and analyzed for EP2 and EP4 mRNA by real-time PCR and protein by western blotting. Data represent the mean ± SEM from three experiments. *, statistically significant (p≤0.05; 2 vs. 18 months).
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References

    1. Agrawal A, Agrawal S, Cao JN, Su H, Osann K, Gupta S. Altered innate immune functioning of dendritic cells in elderly humans: a role of phosphoinositide 3-kinase-signaling pathway. J Immunol. 2007;178:6912–6922. - PubMed
    1. Beharka AA, Wu D, Serafini M, Meydani SN. Mechanism of vitamin E inhibition of cyclooxygenase activity in macrophages from old mice: role of peroxynitrite. Free Radic Biol Med. 2002;32:503–511. - PubMed
    1. Bettelli E, Korn T, Kuchroo VK. Th17: the third member of the effector T cell trilogy. Curr Opin Immunol. 2007;19:652–657. - PMC - PubMed
    1. Bettelli E, Korn T, Oukka M, Kuchroo VK. Induction and effector functions of T(H)17 cells. Nature. 2008;453:1051–1057. - PMC - PubMed
    1. Betz M, Fox BS. Prostaglandin E2 inhibits production of Th1 lymphokines but not of Th2 lymphokines. J Immunol. 1991;146:108–113. - PubMed

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