Immune profile differences between chronic GVHD and late acute GVHD: results of the ABLE/PBMTC 1202 studies

Abstract

Human graft-versus-host disease (GVHD) biology beyond 3 months after hematopoietic stem cell transplantation (HSCT) is complex. The Applied Biomarker in Late Effects of Childhood Cancer study (ABLE/PBMTC1202, NCT02067832) evaluated the immune profiles in chronic GVHD (cGVHD) and late acute GVHD (L-aGVHD). Peripheral blood immune cell and plasma markers were analyzed at day 100 post-HSCT and correlated with GVHD diagnosed according to the National Institutes of Health consensus criteria (NIH-CC) for cGVHD. Of 302 children enrolled, 241 were evaluable as L-aGVHD, cGVHD, active L-aGVHD or cGVHD, and no cGVHD/L-aGVHD. Significant marker differences, adjusted for major clinical factors, were defined as meeting all 3 criteria: receiver-operating characteristic area under the curve ≥0.60, P ≤ .05, and effect ratio ≥1.3 or ≤0.75. Patients with only distinctive features but determined as cGVHD by the adjudication committee (non-NIH-CC) had immune profiles similar to NIH-CC. Both cGVHD and L-aGVHD had decreased transitional B cells and increased cytolytic natural killer (NK) cells. cGVHD had additional abnormalities, with increased activated T cells, naive helper T (Th) and cytotoxic T cells, loss of CD56bright regulatory NK cells, and increased ST2 and soluble CD13. Active L-aGVHD before day 114 had additional abnormalities in naive Th, naive regulatory T (Treg) cell populations, and cytokines, and active cGVHD had an increase in PD-1− and a decrease in PD-1+ memory Treg cells. Unsupervised analysis appeared to show a progression of immune abnormalities from no cGVHD/L-aGVHD to L-aGVHD, with the most complex pattern in cGVHD. Comprehensive immune profiling will allow us to better understand how to minimize L-aGVHD and cGVHD. Further confirmation in adult and pediatric cohorts is needed.

“Age Related Differences in the Biology of Chronic Graft-Versus-Host Disease After Hematopoietic Stem Cell Transplantation”

“…The remaining cytokines were measured by standard colorimetric ELISA (RnD Systems, Minneapolis, US). We found a high accuracy, reproducibility, and linearity for all assays measuring soluble biomarkers and a high stability of analytes upon 24 shipment as have been previously described in adults (4) and children (8).…”

“…Centers assessed children for cGvHD according to the 2005 National Institutes of Health Consensus Criteria, with review by a central study adjudication committee composed of experts in cGvHD. Samples were evaluated by a broad immunophenotyping and plasma cytokine strategy (8).…”

“…FCS files were analyzed using Kaluza software v2 (Beckman Coulter, INC. Mississauga, Canada). Flow cytometry accuracy, reproducibility was ensured by the detailed approaches as previously described (8).…”

“…Samples were collected and shipped as previously described (4,8). Platelet depleted plasmas were isolated and frozen within 24 h of collection, as previously described (4).…”

“…Previous evaluations of cellular and plasma markers of cGvHD in adults, by our group and others, have identified 3 primary cellular populations that characterize cGvHD, including CD21 lo B cells, NK reg cells, and naïve T cells; as well as consistent changes in ST2 (2)(3)(4)(5)(6). Our group recently concluded the Applied Biomarkers of Late Effects of Childhood Cancer (ABLE)/Pediatric Blood and Marrow Transplant Consortium (PBMTC) 1,202 study, which included 302 pediatric transplant patients, 52 of whom had cGvHD (7,8). This has allowed us to compare and contrast potential differences between children and adults in the immune cell and plasma cytokine profiles seen in our previous adults studies that correlate with the development of cGvHD.…”