Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2013 Apr 1;190(7):3225-34.
doi: 10.4049/jimmunol.1200141. Epub 2013 Mar 1.

Independent and Interdependent Immunoregulatory Effects of IL-27, IFN-β, and IL-10 in the Suppression of Human Th17 Cells and Murine Experimental Autoimmune Encephalomyelitis

Affiliations
Free PMC article

Independent and Interdependent Immunoregulatory Effects of IL-27, IFN-β, and IL-10 in the Suppression of Human Th17 Cells and Murine Experimental Autoimmune Encephalomyelitis

Denise C Fitzgerald et al. J Immunol. .
Free PMC article

Abstract

IFN-β, IL-27, and IL-10 have been shown to exert a range of similar immunoregulatory effects in murine and human experimental systems, particularly in Th1- and Th17-mediated models of autoimmune inflammatory disease. In this study we sought to translate some of our previous findings in murine systems to human in vitro models and delineate the interdependence of these different cytokines in their immunoregulatory effects. We demonstrate that human IL-27 upregulates IL-10 in T cell-activated PBMC cultures and that IFN-β drives IL-27 production in activated monocytes. IFN-β-driven IL-27 is responsible for the upregulation of IL-10, but not IL-17 suppression, by IFN-β in human PBMCs. Surprisingly, IL-10 is not required for the suppression of IL-17 by either IL-27 or IFN-β in this model or in de novo differentiating Th17 cells, nor is IL-27 signaling required for the suppression of experimental autoimmune encephalomyelitis (EAE) by IFN-β in vivo. Furthermore, and even more surprisingly, IL-10 is not required for the suppression of Th17-biased EAE by IL-27, in sharp contrast to Th1-biased EAE. In conclusion, IFN-β and IL-27 both induce human IL-10, both suppress human Th17 responses, and both suppress murine EAE. However, IL-27 signaling is not required for the therapeutic effect of IFN-β in EAE. Suppression of Th17-biased EAE by IL-27 is IL-10-independent, in contrast to its mechanism of action in Th1-biased EAE. Taken together, these findings delineate a complex set of interdependent and independent immunoregulatory mechanisms of IFN-β, IL-27, and IL-10 in human experimental models and in murine Th1- and Th17-driven autoimmunity.

Figures

Figure 1
Figure 1. IL-27 upregulates IL-10 production by human PBMCs in vitro
PBMCs from healthy donors were activated with anti-CD2/anti-CD3/anti-CD28 antibodies for 5 days and supernatants were assayed for IL-10 by ELISA. IL-27 upregulated IL-10 production in PBMC cultures (A) but not in purified CD4+ T cells (B). Addition of FBS to serum-free X-VIVO 15 medium potentiated IL-10 production in whole PBMC cultures (C). Data representative of at least two independent experiments (C) or 5–7 donors (A, B).
Figure 2
Figure 2. Endogenous IL-27 drives IL-10 production in human PBMCs in vitro
PBMCs from healthy donors were activated with anti-CD2/anti-CD3/anti-CD28 antibodies for up to 7 days (A) or 5 days (B, C, D) and supernatants were assayed for IL-10 by ELISA (A) Time course analysis shows maximal IL-10 upregulation at 120hrs (n = 4 from each of 3 donors). Anti-IL-27 neutralizing antibody inhibits IL-10 production in the presence of exogenous IL-27 (B) or in basal conditions (C). Exogenous IFN-β also upregulated IL-10 production (D). n = 4–7 donors.
Figure 3
Figure 3. IFN-β upregulates IL-27 production in human monocyte cultures
CD14+ monocytes were purified form PBMCs by immunomagnetic separation and activated with LPS (A) or PGN (B) in the presence or absence of IFN-β for 2 days. Supernatants were assayed for heterodimeric IL-27 by ELISA. PBMCs were activated with anti-CD2/anti-CD3/anti-CD28 in the presence or absence of IFN-β and neutralizing anti-IL-27 antibody for 5 days and supernatants were assayed for IL-10 by ELISA. n = 3–5 donors, data representative of at least 2 experiments.
Figure 4
Figure 4. IFN-β inhibits clinical EAE independent of IL-27 signaling
WT and IL-27R-deficient mice were immunized to develop EAE and treated with IFN-β (50,000U/mouse/day i.p.) from day 0–19 post-immunization. IFN-β effectively suppressed clinical EAE in WT (A; n=8) and il-27rα−/− (B; n=6) mice. Cumulative (C) and mean (D) clinical scores were significantly reduced in WT mice treated with IFN-β and a similar but not quite significant trend in il-27rα−/− mice was observed. N = 6–8 mice per experiment and data are representative of two independent experiments.
Figure 5
Figure 5. Inhibition of Th17 development by IFN-β and IL-27
Naïve CD4+ T cells from cord blood were activated with anti-CD2/anti-CD3/anti-CD28 in the presence or absence of IFN-β (A) or IL-27 (C) in a range of Th17 polarizing cytokine cocktails and IL-17 detection in supernatants was suppressed in all conditions (n=3). Expression of IL-17 by PBMCs from healthy adult donors activated as above was also suppressed by IFN-β (B) and IL-27 as determined by flow cytometry (D, E) and ELISA (F). Neutralization of endogenous IL-27 increased IL-17 production in activated adult PBMC cultures (G) but did not overcome the suppressive effect of IFN-β on IL-17 production by PBMCs (H). Data representative of at least two and up to four independent experimens.
Figure 6
Figure 6. Suppression of Th17 differentiation by IFN-β and IL-27 and of Th17-driven EAE by IL-27 does not require IL-10
Naïve CD4+ T cells from cord blood were activated with anti-CD2/anti-CD3/anti-CD28 in the presence or absence of IFN-β (A) or IL-27 (B) in a range of Th17 polarizing cytokine cocktails +/− neutralizing anti-IL-10 and IL-17 in supernatants was measured by ELISA (n = 3). (C–G) Splenocytes and lymph node cells from immunized donor WT and IL-10-deficient mice were reactivated with MOG35-55 and IL-23 (10 ng/ml) in the presence or absence of IL-27 (10 ng/ml) for 3 days prior to purification of CD4+ cell by immunomagnetic separation and i.v. injection to naïve recipient WT C57BL/6 mice. Recipient animals were treated with PT on days 0 and 2 post transfer (200ng/mouse/day) and clinical EAE was scored daily. Data are pooled from 4 independent experiments (see table 2).

Similar articles

See all similar articles

Cited by 20 articles

See all "Cited by" articles

Publication types

MeSH terms

Feedback