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 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

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 4

GVHD macrophages activate allogeneic T cells.

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GVHD macrophages activate allogeneic T cells. (A) Proliferation of allog...
(A) Proliferation of allogeneic CD4+ and CD8+ cells estimated by CFSE dilution after coculture with DC and macrophage subsets sorted from GVHD or healthy controls. (B) Summary of T cell proliferation (percentage of CFSE dilution) and activation (percentage of CD69+CD8+ T cells and percentage of HLA-DR+ CD4+ T cells) from n = 3 experiments. *P < 0.05, unpaired t test. (C) Heatmap of genes differentially expressed between CD11c+CD14+ monocyte-derived macrophages sorted from healthy control skin (n = 4) and GVHD skin (n = 3) with fold difference in log2 gene expression of greater than 1.3. P < 0.05, unpaired t test. Annotations show functional attributes of genes (based on Entrez Gene summaries) upregulated in GVHD macrophages. (D) CD11c+CD14+ monocyte–derived macrophages sorted from GVHD (n = 3) and healthy control dermis (n = 4) were stimulated with LPS in culture over 10 hours. Chemokine and cytokine production were quantified in supernatants by Luminex assay. Data are represented as mean ± SEM. *P < 0.05; ***P < 0.001, unpaired t test.