Donor monocyte–derived macrophages promote human acute graft-versus-host disease
Laura Jardine, … , A.J. Simpson, Matthew Collin
Laura Jardine, … , A.J. Simpson, Matthew Collin
Published September 1, 2020; First published May 26, 2020
Citation Information: J Clin Invest. 2020;130(9):4574-4586. https://doi.org/10.1172/JCI133909.
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Research Article Immunology Transplantation

Donor monocyte–derived macrophages promote human acute graft-versus-host disease

Abstract

Myelopoiesis is invariably present and contributes to pathology in animal models of graft-versus-host disease (GVHD). In humans, a rich inflammatory infiltrate bearing macrophage markers has also been described in histological studies. In order to determine the origin, functional properties, and role in pathogenesis of these cells, we isolated single-cell suspensions from acute cutaneous GVHD and subjected them to genotype, transcriptome, and in vitro functional analysis. A donor-derived population of CD11c+CD14+ cells was the dominant population of all leukocytes in GVHD. Surface phenotype and NanoString gene expression profiling indicated the closest steady-state counterpart of these cells to be monocyte-derived macrophages. In GVHD, however, there was upregulation of monocyte antigens SIRPα and S100A8/9 transcripts associated with leukocyte trafficking, pattern recognition, antigen presentation, and costimulation. Isolated GVHD macrophages stimulated greater proliferation and activation of allogeneic T cells and secreted higher levels of inflammatory cytokines than their steady-state counterparts. In HLA-matched mixed leukocyte reactions, we also observed differentiation of activated macrophages with a similar phenotype. These exhibited cytopathicity to a keratinocyte cell line and mediated pathological damage to skin explants independently of T cells. Together, these results define the origin, functional properties, and potential pathogenic roles of human GVHD macrophages.

Authors

Laura Jardine, Urszula Cytlak, Merry Gunawan, Gary Reynolds, Kile Green, Xiao-Nong Wang, Sarah Pagan, Maharani Paramitha, Christopher A. Lamb, Anna K. Long, Erin Hurst, Smeera Nair, Graham H. Jackson, Amy Publicover, Venetia Bigley, Muzlifah Haniffa, A.J. Simpson, Matthew Collin

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Figure 1

Mononuclear infiltrates in GVHD contain abundant CD14+CD11c+ myeloid cells.

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Mononuclear infiltrates in GVHD contain abundant CD14+CD11c+ myeloid cel...
Microscopic and flow cytometric evaluation of cutaneous GVHD lesions. (A) Acute GVHD (top row) and healthy control skin (bottom row). Immunohistochemistry with antibodies to CD3, CD11c, CD163, and factor XIIIa (red chromagen) costained with antibody to Ki67 (brown chromagen). Scale bar: 100 μm. (B) Whole-mount immunofluorescence of dermis from healthy controls and patients with GVHD, as indicated with antibodies to CD3 (red), CD11c, (green), and FXIIIA (blue). Scale bar: 50 μm. (C) Enzymatically digested dermis analyzed by flow cytometry from patients with GVHD, patients without GVHD (BMT), or healthy controls (HC), as indicated. Starting from CD45+ mononuclear cells (purple gate), HLA-DR+ cells were gated as shown to arrive at CD11c–CD14+ resident macrophages (brown), CD11c+CD14+CD1c– monocyte-macrophages (red), CD11c+CD14+CD1c+ double-positive cells (pink), CD1c+CD14– cDC2 (cyan), and CD141+ cDC1 (yellow; from the CD14–CD11c– gate). Representative samples of more than 60 experiments are shown. (D) Quantification of digested dermal mononuclear cells from patients with GVHD (n = 39), patients without GVHD (n = 16), or healthy controls (n = 21) as percentages of live cells. Mean + SEM for each group is shown. Groups were compared by 1-way ANOVA, and P values from Tukey’s multiple comparisons tests are shown. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. (E) Ratio of CD11c+CD14+ cells to CD1c+CD14– cells in digests of GVHD, BMT control, or healthy control dermis (14:1c ratio). Median and interquartile range for each group are shown. Groups were compared by Kruskal-Wallis test, and P values from Dunn’s multiple comparisons test are shown. (F) ROC curve analysis of 14:1c ratio in digested cells from GVHD versus BMT controls. AUC = 0.85. Maximal sensitivity and specificity occurred at a ratio of greater than 0.55.