Limits...
An IL-27/NFIL3 signalling axis drives Tim-3 and IL-10 expression and T-cell dysfunction.

Zhu C, Sakuishi K, Xiao S, Sun Z, Zaghouani S, Gu G, Wang C, Tan DJ, Wu C, Rangachari M, Pertel T, Jin HT, Ahmed R, Anderson AC, Kuchroo VK - Nat Commun (2015)

Bottom Line: IL-27-conditioned T helper 1 cells exhibit reduced effector function and are poor mediators of intestinal inflammation.This inhibitory effect is NFIL3 dependent.Thus, our data identify an IL-27/NFIL3 signalling axis as a key regulator of effector T-cell responses via induction of Tim-3, IL-10 and T-cell dysfunction.

View Article: PubMed Central - PubMed

Affiliation: Evergrande Center for Immunologic Diseases, Ann Romney Center for Neurologic Diseases, Harvard Medical School and Brigham and Women's Hospital, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA.

ABSTRACT
The inhibitory receptor T-cell immunoglobulin and mucin domain-3 (Tim-3) has emerged as a critical regulator of the T-cell dysfunction that develops in chronic viral infections and cancers. However, little is known regarding the signalling pathways that drive Tim-3 expression. Here, we demonstrate that interleukin (IL)-27 induces nuclear factor, interleukin 3 regulated (NFIL3), which promotes permissive chromatin remodelling of the Tim-3 locus and induces Tim-3 expression together with the immunosuppressive cytokine IL-10. We further show that the IL-27/NFIL3 signalling axis is crucial for the induction of Tim-3 in vivo. IL-27-conditioned T helper 1 cells exhibit reduced effector function and are poor mediators of intestinal inflammation. This inhibitory effect is NFIL3 dependent. In contrast, tumour-infiltrating lymphocytes from IL-27R(-/-) mice exhibit reduced NFIL3, less Tim-3 expression and failure to develop dysfunctional phenotype, resulting in better tumour growth control. Thus, our data identify an IL-27/NFIL3 signalling axis as a key regulator of effector T-cell responses via induction of Tim-3, IL-10 and T-cell dysfunction.

No MeSH data available.


Related in: MedlinePlus

Ectopic expression of NFIL3 in CD4+ T cells attenuates gut pathology in adoptively transferred enteritis/colitis. Naïve CD4+ T cells from C57BL/6 mice were transduced with NFIL3-expressing retrovirus (NFIL3) or control empty retrovirus (GFP). Cells were injected i.p. into Rag1−/− recipient mice to induce gut inflammation (0.7 × 107 cells/mouse). (a) Wasting disease was monitored for 10 weeks after transfer. Statistical analysis was performed on the combination of total animals from two independent experiments. Data are shown as mean ± SEM. Mann Whitney test two-tailed P=0.0064. (b) Immunopathology of gut inflammation. Two randomly selected tissue sections from each animal were graded for crypt damage and T cell infiltration. Two scores were given to each animal to represent the overall gut immunopathology. Data are shown as mean ± SEM. Unpaired t test two-tailed P=0.0056. (c) The recipient mice were sacrificed 6 weeks after T cell transfer for ex vivo analysis of cytokine production (IL-2, IL-10, and IFN-γ) and Tim-3 expression by flow cytometry. (d) Naïve CD4+ T cells from WT FoxP3-GFP KI and NFIL3−/− x FoxP3-GFP KI mice (CD4+CD62L+GFP−) were differentiated into Th1 cells with or without the presence of IL-27. On day 6, GFP+ (FoxP3+) cells were isolated by cell sorting and were injected i.p. into Rag1−/− recipients to induce colitis. Colon pathology was analyzed 8 weeks after transfer. Each score represents the pathology from one recipient mouse (NFIL3−/− Th1 n=8; NFIL3−/− Th1+IL-27 n=7; WT Th1 n=7; WT Th1+IL-27 n=9. Data are shown as mean ± SEM, t test).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4311884&req=5

Figure 6: Ectopic expression of NFIL3 in CD4+ T cells attenuates gut pathology in adoptively transferred enteritis/colitis. Naïve CD4+ T cells from C57BL/6 mice were transduced with NFIL3-expressing retrovirus (NFIL3) or control empty retrovirus (GFP). Cells were injected i.p. into Rag1−/− recipient mice to induce gut inflammation (0.7 × 107 cells/mouse). (a) Wasting disease was monitored for 10 weeks after transfer. Statistical analysis was performed on the combination of total animals from two independent experiments. Data are shown as mean ± SEM. Mann Whitney test two-tailed P=0.0064. (b) Immunopathology of gut inflammation. Two randomly selected tissue sections from each animal were graded for crypt damage and T cell infiltration. Two scores were given to each animal to represent the overall gut immunopathology. Data are shown as mean ± SEM. Unpaired t test two-tailed P=0.0056. (c) The recipient mice were sacrificed 6 weeks after T cell transfer for ex vivo analysis of cytokine production (IL-2, IL-10, and IFN-γ) and Tim-3 expression by flow cytometry. (d) Naïve CD4+ T cells from WT FoxP3-GFP KI and NFIL3−/− x FoxP3-GFP KI mice (CD4+CD62L+GFP−) were differentiated into Th1 cells with or without the presence of IL-27. On day 6, GFP+ (FoxP3+) cells were isolated by cell sorting and were injected i.p. into Rag1−/− recipients to induce colitis. Colon pathology was analyzed 8 weeks after transfer. Each score represents the pathology from one recipient mouse (NFIL3−/− Th1 n=8; NFIL3−/− Th1+IL-27 n=7; WT Th1 n=7; WT Th1+IL-27 n=9. Data are shown as mean ± SEM, t test).

Mentions: Our data indicate that IL-27-induced NFIL3 plays a key role in the induction of Tim-3 and IL-10 in effector T cells and increases dysfunctional phenotype in effector CD4+ T cells. We therefore hypothesized that overexpression of NFIL3 could dampen effector T cell function. Thus, we transduced naïve CD4+ T cells with NFIL3-expressing retrovirus (NFIL3) and transferred these cells into Rag1−/−13 recipient mice and observed these mice for the development of gut inflammation. Recipient mice that received NFIL3-transduced CD4+ T cells failed to develop wasting disease 10 weeks after transfer (Fig. 6a), when compared with the control mice that received empty virus-transduced (GFP) CD4+ T cells. Histological analysis further revealed that empty virus-transduced T cells induced significantly higher mucosal inflammation than NFIL3 transduced T cells. After grading the crypt damage and cell infiltration (as described in Supplementary Fig. 8a), we observed that almost all of the recipient mice that received empty virus-transduced CD4+ T cells developed clear histological signs of both enteritis and colitis. In contrast, about half of the mice transferred with NFIL3-transduced T cells did not have any signs of gut inflammation and the rest of the mice had a milder inflammation when compared to the mice transferred with control empty virus-transduced T cells (Fig. 6b). We also found that NFIL3-transduced T cells were localized in Peyer’s patches, suggesting that failure to induce inflammation in the gut was not due to a defect of cell migration (Supplementary Fig. 8b). In addition, when we harvested empty virus- and NFIL3-transduced cells from the mesenteric lymph nodes of recipient mice 6 weeks post transfer, we found that NFIL3-transduced cells exhibited high Tim-3 expression and IL-10 production, but low IFN-γ production. Although not statistically significant, a clear trend towards low IL-2 production was also observed (Fig. 6c). Collectively, these data indicate that the effector function of NFIL3-transduced CD4+ T cells was suppressed in vivo, thereby attenuating the induction of gut inflammation.


An IL-27/NFIL3 signalling axis drives Tim-3 and IL-10 expression and T-cell dysfunction.

Zhu C, Sakuishi K, Xiao S, Sun Z, Zaghouani S, Gu G, Wang C, Tan DJ, Wu C, Rangachari M, Pertel T, Jin HT, Ahmed R, Anderson AC, Kuchroo VK - Nat Commun (2015)

Ectopic expression of NFIL3 in CD4+ T cells attenuates gut pathology in adoptively transferred enteritis/colitis. Naïve CD4+ T cells from C57BL/6 mice were transduced with NFIL3-expressing retrovirus (NFIL3) or control empty retrovirus (GFP). Cells were injected i.p. into Rag1−/− recipient mice to induce gut inflammation (0.7 × 107 cells/mouse). (a) Wasting disease was monitored for 10 weeks after transfer. Statistical analysis was performed on the combination of total animals from two independent experiments. Data are shown as mean ± SEM. Mann Whitney test two-tailed P=0.0064. (b) Immunopathology of gut inflammation. Two randomly selected tissue sections from each animal were graded for crypt damage and T cell infiltration. Two scores were given to each animal to represent the overall gut immunopathology. Data are shown as mean ± SEM. Unpaired t test two-tailed P=0.0056. (c) The recipient mice were sacrificed 6 weeks after T cell transfer for ex vivo analysis of cytokine production (IL-2, IL-10, and IFN-γ) and Tim-3 expression by flow cytometry. (d) Naïve CD4+ T cells from WT FoxP3-GFP KI and NFIL3−/− x FoxP3-GFP KI mice (CD4+CD62L+GFP−) were differentiated into Th1 cells with or without the presence of IL-27. On day 6, GFP+ (FoxP3+) cells were isolated by cell sorting and were injected i.p. into Rag1−/− recipients to induce colitis. Colon pathology was analyzed 8 weeks after transfer. Each score represents the pathology from one recipient mouse (NFIL3−/− Th1 n=8; NFIL3−/− Th1+IL-27 n=7; WT Th1 n=7; WT Th1+IL-27 n=9. Data are shown as mean ± SEM, t test).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4311884&req=5

Figure 6: Ectopic expression of NFIL3 in CD4+ T cells attenuates gut pathology in adoptively transferred enteritis/colitis. Naïve CD4+ T cells from C57BL/6 mice were transduced with NFIL3-expressing retrovirus (NFIL3) or control empty retrovirus (GFP). Cells were injected i.p. into Rag1−/− recipient mice to induce gut inflammation (0.7 × 107 cells/mouse). (a) Wasting disease was monitored for 10 weeks after transfer. Statistical analysis was performed on the combination of total animals from two independent experiments. Data are shown as mean ± SEM. Mann Whitney test two-tailed P=0.0064. (b) Immunopathology of gut inflammation. Two randomly selected tissue sections from each animal were graded for crypt damage and T cell infiltration. Two scores were given to each animal to represent the overall gut immunopathology. Data are shown as mean ± SEM. Unpaired t test two-tailed P=0.0056. (c) The recipient mice were sacrificed 6 weeks after T cell transfer for ex vivo analysis of cytokine production (IL-2, IL-10, and IFN-γ) and Tim-3 expression by flow cytometry. (d) Naïve CD4+ T cells from WT FoxP3-GFP KI and NFIL3−/− x FoxP3-GFP KI mice (CD4+CD62L+GFP−) were differentiated into Th1 cells with or without the presence of IL-27. On day 6, GFP+ (FoxP3+) cells were isolated by cell sorting and were injected i.p. into Rag1−/− recipients to induce colitis. Colon pathology was analyzed 8 weeks after transfer. Each score represents the pathology from one recipient mouse (NFIL3−/− Th1 n=8; NFIL3−/− Th1+IL-27 n=7; WT Th1 n=7; WT Th1+IL-27 n=9. Data are shown as mean ± SEM, t test).
Mentions: Our data indicate that IL-27-induced NFIL3 plays a key role in the induction of Tim-3 and IL-10 in effector T cells and increases dysfunctional phenotype in effector CD4+ T cells. We therefore hypothesized that overexpression of NFIL3 could dampen effector T cell function. Thus, we transduced naïve CD4+ T cells with NFIL3-expressing retrovirus (NFIL3) and transferred these cells into Rag1−/−13 recipient mice and observed these mice for the development of gut inflammation. Recipient mice that received NFIL3-transduced CD4+ T cells failed to develop wasting disease 10 weeks after transfer (Fig. 6a), when compared with the control mice that received empty virus-transduced (GFP) CD4+ T cells. Histological analysis further revealed that empty virus-transduced T cells induced significantly higher mucosal inflammation than NFIL3 transduced T cells. After grading the crypt damage and cell infiltration (as described in Supplementary Fig. 8a), we observed that almost all of the recipient mice that received empty virus-transduced CD4+ T cells developed clear histological signs of both enteritis and colitis. In contrast, about half of the mice transferred with NFIL3-transduced T cells did not have any signs of gut inflammation and the rest of the mice had a milder inflammation when compared to the mice transferred with control empty virus-transduced T cells (Fig. 6b). We also found that NFIL3-transduced T cells were localized in Peyer’s patches, suggesting that failure to induce inflammation in the gut was not due to a defect of cell migration (Supplementary Fig. 8b). In addition, when we harvested empty virus- and NFIL3-transduced cells from the mesenteric lymph nodes of recipient mice 6 weeks post transfer, we found that NFIL3-transduced cells exhibited high Tim-3 expression and IL-10 production, but low IFN-γ production. Although not statistically significant, a clear trend towards low IL-2 production was also observed (Fig. 6c). Collectively, these data indicate that the effector function of NFIL3-transduced CD4+ T cells was suppressed in vivo, thereby attenuating the induction of gut inflammation.

Bottom Line: IL-27-conditioned T helper 1 cells exhibit reduced effector function and are poor mediators of intestinal inflammation.This inhibitory effect is NFIL3 dependent.Thus, our data identify an IL-27/NFIL3 signalling axis as a key regulator of effector T-cell responses via induction of Tim-3, IL-10 and T-cell dysfunction.

View Article: PubMed Central - PubMed

Affiliation: Evergrande Center for Immunologic Diseases, Ann Romney Center for Neurologic Diseases, Harvard Medical School and Brigham and Women's Hospital, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA.

ABSTRACT
The inhibitory receptor T-cell immunoglobulin and mucin domain-3 (Tim-3) has emerged as a critical regulator of the T-cell dysfunction that develops in chronic viral infections and cancers. However, little is known regarding the signalling pathways that drive Tim-3 expression. Here, we demonstrate that interleukin (IL)-27 induces nuclear factor, interleukin 3 regulated (NFIL3), which promotes permissive chromatin remodelling of the Tim-3 locus and induces Tim-3 expression together with the immunosuppressive cytokine IL-10. We further show that the IL-27/NFIL3 signalling axis is crucial for the induction of Tim-3 in vivo. IL-27-conditioned T helper 1 cells exhibit reduced effector function and are poor mediators of intestinal inflammation. This inhibitory effect is NFIL3 dependent. In contrast, tumour-infiltrating lymphocytes from IL-27R(-/-) mice exhibit reduced NFIL3, less Tim-3 expression and failure to develop dysfunctional phenotype, resulting in better tumour growth control. Thus, our data identify an IL-27/NFIL3 signalling axis as a key regulator of effector T-cell responses via induction of Tim-3, IL-10 and T-cell dysfunction.

No MeSH data available.


Related in: MedlinePlus