Cytokine and Chemokine Patterns Across 100 Days after Hematopoietic Stem Cell Transplantation in Children Joseph DiCarlo, Rajni Agarwal-Hashmi, Ami Shah,

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Cytokine and Chemokine Patterns Across 100 Days after Hematopoietic Stem Cell Transplantation in Children Joseph DiCarlo, Rajni Agarwal-Hashmi, Ami Shah, Paul Kim, Laila Craveiro, Renna Killen, Yael Rosenberg-Hasson, Holden Maecker Biology of Blood and Marrow Transplantation Volume 20, Issue 3, Pages 361-369 (March 2014) DOI: 10.1016/j.bbmt.2013.11.026 Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 1 WBC count (y-axis; cells per microliter) from all study patients (N = 51) plotted against number of weeks after transplantation (x-axis). Total WBC count rebounded more rapidly in children who developed GVHD or VOD. Biology of Blood and Marrow Transplantation 2014 20, 361-369DOI: (10.1016/j.bbmt.2013.11.026) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 2 Data from children who did not develop complications after HSCT (n = 33), depicting several cytokines in very low concentrations compared with normal control subjects. This graph depicts each cytokine MFI as a multiple of the concurrent control value (ie, “control” is the horizontal line at y = 1, whereas individual cytokine MFIs are depicted either above or below that reference; eg, .2 would represent a cytokine secreted at 20% of control rate). MFI for many cytokines was well below that of concurrently measured control subjects. Biology of Blood and Marrow Transplantation 2014 20, 361-369DOI: (10.1016/j.bbmt.2013.11.026) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 3 Further data from the same children who did not develop complications after HSCT (n = 33), depicting the few cytokines present in concentrations greater than in concurrent control subjects. Each cytokine MFI is depicted as a multiple of the concurrent control value (ie, “control” is the horizontal line at y = 1, whereas individual cytokine MFIs are depicted as multiples of the control value; we measured MFI values as high as 15 × control baseline, whereas values below “1” denotes decreased secretion; eg, “.2” would represent a cytokine secreted at 20% of control rate). Biology of Blood and Marrow Transplantation 2014 20, 361-369DOI: (10.1016/j.bbmt.2013.11.026) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 4 All transplant recipients exhibited increased MFI for a limited number of cytokines. Recipients who developed VOD had HGF and IL-6 activity that was significantly higher than in recipients without complications. IP-10 and MCP-1 varied greatly between subjects but not between groups (noncomplication versus GVHD versus VOD). Biology of Blood and Marrow Transplantation 2014 20, 361-369DOI: (10.1016/j.bbmt.2013.11.026) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 5 Cytokine profile in children who developed GVHD (n = 10). Increases in HGF and IL-6 activity were not as high as in recipients with VOD. Biology of Blood and Marrow Transplantation 2014 20, 361-369DOI: (10.1016/j.bbmt.2013.11.026) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 6 Median fluorescence activity. Number of weeks after transplantation along the x-axis, with day of transplant samples collected at x = 1. Patients without complications (n = 33, MFI – SD) dark gray band in foreground; GVHD (n = 10, MFI – SD) light gray with black outline; VOD (n = 5) (MFI + SD) medium gray band in background. Biology of Blood and Marrow Transplantation 2014 20, 361-369DOI: (10.1016/j.bbmt.2013.11.026) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 7 Receiver operating characteristic curves were constructed for prediction of vaso-occlusive disease by MFI measured 21 days after transplantation. HGF, GRO-α, and MCP-1 were superior to IL-6. Biology of Blood and Marrow Transplantation 2014 20, 361-369DOI: (10.1016/j.bbmt.2013.11.026) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 8 Receiver operating characteristic curves were constructed for prediction of GVHD by MFI measured 21 days after transplantation. HGF and GRO-α demonstrated acceptable sensitivity but poor specificity. Biology of Blood and Marrow Transplantation 2014 20, 361-369DOI: (10.1016/j.bbmt.2013.11.026) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions