Intestinal epithelial Ceacam1 deficiency prevents steroid-refractory acute gut graft-versus-host disease
Qingxiao Song, Moqian Zheng, Qinjian Li, Xiwei Wu, Boxi Lin, Tae Hyuk Kang, Hanjun Qin, Maciej Kujawski, Raju K. Pillai, James L. Lin, Ryotaro Nakamura, John Shively, Paul J. Martin, Defu Zeng
Qingxiao Song, Moqian Zheng, Qinjian Li, Xiwei Wu, Boxi Lin, Tae Hyuk Kang, Hanjun Qin, Maciej Kujawski, Raju K. Pillai, James L. Lin, Ryotaro Nakamura, John Shively, Paul J. Martin, Defu Zeng
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Research Article Hematology Immunology Transplantation

Intestinal epithelial Ceacam1 deficiency prevents steroid-refractory acute gut graft-versus-host disease

Abstract

Steroid-refractory gut acute graft-versus-host disease (SR-Gut-aGVHD) is the major cause of nonrelapse death after allogeneic hematopoietic cell transplantation. High numbers of donor-type IL-22+ T cells, IL-22–dependent dysbiosis, and loss of antiinflammatory CX3CR1hi mononuclear phagocytes (MNPs) play critical roles in SR-Gut-aGVHD pathogenesis. CEACAM1 on intestinal epithelial cells (IECs) is proposed to regulate bacterial translocation and subsequent immune responses in the intestine. Here, with imaging mass cytometry (IMC), combined scRNA-Seq with ATAC-Seq, and high-dimensional flow cytometry analysis, we show that CEACAM1 expression was enhanced on IECs in murine and human SR-Gut-aGVHD. Ceacam1 deficiency on host IECs effectively prevented SR-Gut-aGVHD in murine models. Ceacam1 deficiency on IECs resulted in (i) higher numbers of IL-22+IL-10+Foxp3+CD4+ peripheral Tregs (pTregs) and lower numbers of conventional IL-22+CD4+ T (Tcon), Th/Tc1, and Th17 cells in the intestine; (ii) higher prevalence of beneficial commensal bacteria that augment colonic pTreg expansion, with lower prevalence of pathogenic bacteria; and (iii) higher numbers of antiinflammatory CD103–CX3CR1hi MNPs that produce indoleamine 2,3-dioxygenase (IDO) and IL-10, with lower numbers of proinflammatory CD103+CX3CR1lo MNPs that produce IL-6. Thus, specifically targeting IEC CEACAM1 represents a promising approach for prevention of SR-Gut-aGVHD.

Authors

Qingxiao Song, Moqian Zheng, Qinjian Li, Xiwei Wu, Boxi Lin, Tae Hyuk Kang, Hanjun Qin, Maciej Kujawski, Raju K. Pillai, James L. Lin, Ryotaro Nakamura, John Shively, Paul J. Martin, Defu Zeng

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Figure 9

Amelioration of SR-Gut-aGVHD by Ceacam1 deficiency in IECs is associated with higher numbers of antiinflammatory CD103–CX3CR1hi MNPs and lower numbers of proinflammatory CD103+CX3CR1lo MNPs in the colon.

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Amelioration of SR-Gut-aGVHD by Ceacam1 deficiency in IECs is associated...
Lethally irradiated WT and IEC-Ceacam1–/– chimeric recipients were engrafted with splenocytes and TCD-BM from WT C57BL/6 donors as described for Figure 2. On day 25, CD103+CX3CR1lo and CD103–CX3CR1hi MNPs in the colons were analyzed, and cytokines were measured in homogenized colon tissue. (A) Representative flow cytometry pattern and gating strategy for CD103+CX3CR1lo and CD103–CX3CR1hi MNP. (B) Plots of mean ± SEM of %CD103+CX3CR1lo and CD103–CX3CR1hi MNPs among MNCs and their respective yields. n = 6–7. (C) Representative flow cytometry pattern and gating strategy showing IDO+IL-10+ population in CD103+CX3CR1lo and CD103–CX3CR1hi MNP. (D) Percentages of CD103+CX3CR1lo and CD103–CX3CR1hi MNPs expressing both IL-10 and IDO. Data are presented as mean ± SEM; n = 6–7. (E and F) Representative flow cytometry patterns and percentages of IL-6 in CD103+CX3CR1lo and CD103–CX3CR1hi MNPs. Data are presented as mean ± SEM; n = 13. Comp, compensation. (G) Concentration of active TGF-β, IFN-γ, IL-6, TNF, and IL-2 in colon tissue homogenate. n = 3–4. (H and I) Representative flow cytometry patterns and percentages of IL-2+CD4+ T cells. n = 12–16. Combined from 2 experiments. P values are shown in B, D, F, G, and I. Unpaired 2-tailed Student’s t test was used to compare 2 means in B, F, and I. Two-way ANOVA was used to compare means in D and G.