title: Citrulline and Monocyte-Derived Macrophage Reactivity before Conditioning Predict Acute Graft-versus-Host Disease.
abstract: During conditioning, intestinal damage induces microbial translocation which primes macrophage reactivity and leads to donor-derived T cell stimulation. Little is known about the role of intestinal health and macrophage reactivity before conditioning in the development of acute graft-versus-host disease (aGVHD) in patients undergoing allogeneic hematopoietic cell transplantation (allo-HCT). We assessed (1) citrulline, a surrogate marker of functional enterocyte mass and (2) circulating monocyte-derived macrophage reactivity, before allo-HCT. Forty-seven consecutive patients were prospectively included. Citrulline levels from blood samples withdrawn 30 days before transplantation were assessed using liquid chromatography combined with mass spectrometry. Monocyte-derived macrophages were isolated and incubated with 5 pathogen-associated molecular patterns: lipopolysaccharide, PamCSK4, flagellin, muramyl dipeptide, and curdlan. Multiplex fluorescent immunoassay on culture supernatant assessed levels of TNF-α, IL-1β, IL-6, and IL-10 in each condition. Citrulline and cytokine levels were analyzed relatively to aGVHD onset within 100 days after transplantation. Citrulline levels were lower in the aGVHD group (n = 20) than in the no-aGVHD group (n = 27) (P = .005). Conversely, IL-6 and IL-10 were greater in aGVHD group, especially after curdlan stimulation (P = .005 and P = .012). Citrulline levels ≤20 µmol/L, IL-6 ≥ 332 pg/mL, and IL-10 ≥ 90 pg/mL were associated with aGVHD development (log-rank test, P = .002, P = .041, and P < .0001, respectively). In multivariate analysis, IL-10 ≥ 90 pg/mL, myeloablative conditioning, and citrulline ≤20 µmol/L remained independent factors of aGVHD development (hazard ratio [HR],  8.18, P = .0003; HR, 4.28, P = .006; and HR, 4.43, P = .01, respectively). Preconditioning citrulline and monocyte-derived macrophage reactivity are objective surrogate markers suitable to identify patients at risk of developing aGVHD. This work highlights the influence of preconditioning status in aGVHD development.
authors.affiliation: LIRIC UMR 995 Inserm, University of Lille, Lille, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Hueso
authors.affiliation: Stem Cell Transplantation Unit, CHU Lille, Lille, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Coiteux
authors.affiliation: Department of Biochemistry and Molecular Biology, CHU Lille, Lille, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Joncquel Chevalier Curt
authors.affiliation: Department of Biostatistics, CHU Lille, Lille, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Labreuche
authors.affiliation: LIRIC UMR 995 Inserm, University of Lille, Lille, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Jouault
authors.affiliation: LIRIC UMR 995 Inserm, University of Lille, Lille, France; Stem Cell Transplantation Unit, CHU Lille, Lille, France.
authors.forename: Ibrahim
authors.initials: I
authors.lastname: Yakoub-Agha
authors.affiliation: LIRIC UMR 995 Inserm, University of Lille, Lille, France; Department of Nutrition, CHU Lille, Lille, France. Electronic address: david.seguy@chru-lille.fr.
authors.forename: David
authors.initials: D
authors.lastname: Seguy
mesh_terms: D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000097:blood; D002956:Citrulline / Q000097:blood*; D005260:Female; D006086:Graft vs Host Disease / Q000175:diagnosis* / Q000209:etiology; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects / Q000379:methods; D006801:Humans; D007410:Intestinal Diseases / Q000139:chemically induced / Q000382:microbiology; D008213:Lymphocyte Activation / Q000276:immunology; D008264:Macrophages / Q000276:immunology* / Q000382:microbiology; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000166:cytology; D011237:Predictive Value of Tests; D019172:Transplantation Conditioning / Q000009:adverse effects / Q000379:methods
chemical_list: D015415:Biomarkers; D002956:Citrulline
keywords: Citrulline; Interleukin; Intestinal damage; Macrophage