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 1

Colonic epithelial expression of CEACAM1 and infiltration of IL-22+IFN-γ+ T cells are enhanced in patients with SR-Gut-GVHD.

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Colonic epithelial expression of CEACAM1 and infiltration of IL-22+IFN-γ...
Using H&E staining and image mass cytometry (IMC), we evaluated the severity of tissue damage indicated by immune cell infiltration and CEACAM1 expression on epithelial cells in colon biopsies from SR-Gut-GVHD (n = 3) and Non–SR-GVHD patients (n = 3). The IMC scan was based on 1 µm per pixel. The H&E overview was acquired withZEISS ZEN Tile Scan, and the magnification of the indicated regions are shown with yellow boxes (scale bars) on the right. (A) Representative IMC staining patterns of CEACAM1, Ki-67, and E-cadherin in patient colon biopsy samples. Scale bar: 200 μm. (B) Percentage of CEACAM1+Ki-67+E-cadherin+ cells among total MNCs. The P value is shown. (C) Representative IMC staining pattern of colon biopsy from a non-SR-Gut-GVHD patient. The panel shows staining for DAPI (blue), CD3 (green), CD8α (cyan), and IL-22 (red). White arrows indicate representative CD8+ T cells. (D) Representative IMC staining patterns for analysis of IL-22+CD4+ and IL-22+CD8+T cells in a colon biopsy sample from a patient with SR-Gut-GVHD. The arrows in the upper rows of the two arrays of panels indicate representative IL-22+CD4+ or IL-22+CD8+ T cells. The lower rows provide a more detailed analysis of T cell subsets from the same patient. The white arrows indicate IL-22+CD8+ T cells with expression of Ahr, IFN-γ, T-bet, or IL-17A, while the green arrows indicate CD4+ T cells with or without expression of Ahr, IFN-γ, T-bet, or IL-17A. (32. AUTHOR: Please edit to specify the total magnifications for the images in C and D.) (E) General cellular distribution in the t-distributed stochastic neighbor embedding (t-SNE) map of all MNCs (left panel) and IL-22+CD4+ and IL-22+CD8+ T cells (right panel). (F) Percentages of IL-22+ T cells among total MNCs (left panel), CD4+ and CD8+ T among IL-22+ T cells in SR-Gut-GVHD, and non-SR-Gut-GVHD tissues. (G) Percentages of Ahr+ and T-bet+ cells among IL-22+ T cells. (H) Percentages of IFN-γ+IL-17A+Ahr+T-bet+IL-22+ T cells among total MNCs. P values were calculated by unpaired 2-tailed Student’s t tests (B, F-H). *P < 0.05; ***P < 0.001.