Figure 1.
Systemic IL-6 dysregulation originates in recipient DCs. (A,C,F-J) Lethally irradiated recipient CD11cCre (intact IL-6 signaling), CD11cCre × IL-6Rfl (DCs are deficient in classical and unable to induce cluster signaling), CD11cCre × IL-6fl (DCs are deficient in IL-6 and unable to induce cluster signaling), and IL-6−/− (all recipient cells are deficient in IL-6) mice received transplants of 5 × 106 T cells from wild-type BALB/c (BALB/c.WT) donors. (B) DCs can signal via IL-6 or induce IL-6 signaling to gp130 on T cells through 3 pathways: classical (involving DC-derived IL-6), trans (requiring DC-derived IL-6 ligated to any source of sIL-6R), and cluster (requiring DC-derived IL-6 and IL-6R). (C) Peripheral blood serum levels of IL-6 from recipients at day 4 and day 7 after transplantation (n = 5-8 mice per group from 2 experiments). (D-E) B6.WT mice were irradiated (1000 cGy), and 8 hours later, CD103+CD11b−, CD103+CD11b+, and CD103−CD11b+ DCs (CD11c+MHCIIhiCD64−F4/80−) and non-DC boolean-gated CD64+F4/80+ macrophages were sort purified from the mLNs (D) and IL6 mRNA levels measured by quantitative polymerase chain reaction and quantified relative to CD103+CD11b− DCs (n = 3, each pooled from mLNs from 2-5 mice) (E). (F) Representative flow cytometry plots of IL-22 and IL-17A expression in CD4+ T cells from the mLNs (concatenated from 4 mice per group) and stacked bar graphs of the frequency (G) and total numbers (H) of donor Th17 (CD4+IL-17A+) and Th22 (CD4+IL-22+IL-17A−) cells in the mLNs and spleen at day 7 posttransplantation (n = 4-14 mice per group from 2-3 experiments). (I-J) Representative flow cytometry plots (I) and frequency (J) of interferon-γ (IFNγ), tumor necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and IL-10 expression in CD4+ T cells from the mLNs (concatenated from 4 mice per group; n = 8-14 mice per group from 3 experiments). Data presented as mean ± standard error of the mean. *P < .05, **P < .01, ***P < .001. †P < .05, ††P < .01, †††P < .001 for Th22. ‡P < .05, ‡‡P < .01, ‡‡‡P < .001 for Th17. MHC, major histocompatibility complex; n.d., not detectable; n.s., not significant; TBI, total-body irradiation.

Systemic IL-6 dysregulation originates in recipient DCs. (A,C,F-J) Lethally irradiated recipient CD11cCre (intact IL-6 signaling), CD11cCre × IL-6Rfl (DCs are deficient in classical and unable to induce cluster signaling), CD11cCre × IL-6fl (DCs are deficient in IL-6 and unable to induce cluster signaling), and IL-6−/− (all recipient cells are deficient in IL-6) mice received transplants of 5 × 106 T cells from wild-type BALB/c (BALB/c.WT) donors. (B) DCs can signal via IL-6 or induce IL-6 signaling to gp130 on T cells through 3 pathways: classical (involving DC-derived IL-6), trans (requiring DC-derived IL-6 ligated to any source of sIL-6R), and cluster (requiring DC-derived IL-6 and IL-6R). (C) Peripheral blood serum levels of IL-6 from recipients at day 4 and day 7 after transplantation (n = 5-8 mice per group from 2 experiments). (D-E) B6.WT mice were irradiated (1000 cGy), and 8 hours later, CD103+CD11b−, CD103+CD11b+, and CD103−CD11b+ DCs (CD11c+MHCIIhiCD64−F4/80−) and non-DC boolean-gated CD64+F4/80+ macrophages were sort purified from the mLNs (D) and IL6 mRNA levels measured by quantitative polymerase chain reaction and quantified relative to CD103+CD11b− DCs (n = 3, each pooled from mLNs from 2-5 mice) (E). (F) Representative flow cytometry plots of IL-22 and IL-17A expression in CD4+ T cells from the mLNs (concatenated from 4 mice per group) and stacked bar graphs of the frequency (G) and total numbers (H) of donor Th17 (CD4+IL-17A+) and Th22 (CD4+IL-22+IL-17A−) cells in the mLNs and spleen at day 7 posttransplantation (n = 4-14 mice per group from 2-3 experiments). (I-J) Representative flow cytometry plots (I) and frequency (J) of interferon-γ (IFNγ), tumor necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and IL-10 expression in CD4+ T cells from the mLNs (concatenated from 4 mice per group; n = 8-14 mice per group from 3 experiments). Data presented as mean ± standard error of the mean. *P < .05, **P < .01, ***P < .001. †P < .05, ††P < .01, †††P < .001 for Th22. ‡P < .05, ‡‡P < .01, ‡‡‡P < .001 for Th17. MHC, major histocompatibility complex; n.d., not detectable; n.s., not significant; TBI, total-body irradiation.

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