Cytomegalovirus Viral Load and Virus-Specific Immune Reconstitution after Peripheral Blood Stem Cell versus Bone Marrow Transplantation Abraham Guerrero,

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Cytomegalovirus Viral Load and Virus-Specific Immune Reconstitution after Peripheral Blood Stem Cell versus Bone Marrow Transplantation Abraham Guerrero, Stanley R. Riddell, Jan Storek, Terry Stevens-Ayers, Barry Storer, John A. Zaia, Stephen Forman, Robert S. Negrin, Thomas Chauncey, William Bensinger, Michael Boeckh Biology of Blood and Marrow Transplantation Volume 18, Issue 1, Pages 66-75 (January 2012) DOI: 10.1016/j.bbmt.2011.05.010 Copyright © 2012 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 1 Differences in CMV infection and disease between BM and PBSC recipients. Time to positive antigenemia or PCR positivity at any level (A), antigenemia >100 positive cells per slide or CMV DNA >100,000 copies/mL plasma (B), and CMV disease (C) among CMV seropositive recipients (BM, n = 50; PBSC, n = 46). Biology of Blood and Marrow Transplantation 2012 18, 66-75DOI: (10.1016/j.bbmt.2011.05.010) Copyright © 2012 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 2 Effect of G-CSF on donor peripheral blood mononuclear cells (PBMC) and stem cell (SC) product. (A) Represents the number of CMV-specific CD4+ T helper cells in donor PBMC and the stem cell product, pre- and post-G-CSF treatment (n = 12). P value is from the Wilcoxon matched pairs test. Biology of Blood and Marrow Transplantation 2012 18, 66-75DOI: (10.1016/j.bbmt.2011.05.010) Copyright © 2012 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 3 Differences in CMV-specific T cell immune reconstitution between BM and PBSC recipients. (A) Represents the proportion of patients with CMV-specific cell lysis greater than 10% between BM and PBSC recipients at day 30 (BM, n = 8; PBSC, n = 8), 80 (BM, n = 9; PBSC, n = 8), 180 (BM, n = 6; PBSC, n = 7), and 365 (BM, n = 6; PBSC, n = 4) posttransplantation. (B) Represents the proportion of patients with ≥33 CMV-specific CD4+ T cells per 106 PBMC as determined by limiting dilution assay at day 30, 80, 180, and 365 (BM, n = 22; PBSC, n = 22) posttransplantation. Biology of Blood and Marrow Transplantation 2012 18, 66-75DOI: (10.1016/j.bbmt.2011.05.010) Copyright © 2012 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 4 Differences in polyfunctional CMV-specific CD8+ T cells between BM and PBSC recipients. (A, B) Represents the absolute number of CD8+ T cells in BM (n = 7†) and PBSC (n = 6†) recipients positive for CD107a, IFN-γ, MIP-1β, and TNF-α at day 30 and day 80 posttransplantation. (C, D) Represents the absolute proportion of polyfunctional CMV-specific CD8+ T cells within BM and PBSC recipients at day 30 and day 80. (E, F) Represents the MFI of IFN-γ based on the degree of decreasing polyfunctionality of CMV-specific CD8+ T cells by the loss of one marker (CD107a, MIP-1β, and TNF-α). Polyfunctional T cells grouped by T cells positive for 1, 2, 3, or 4 of the markers based on the 16 possible combinations according to SPICE. The P value is a result of the Student test. Bars represent the mean value within each group, and error bars are the standard error of the mean. Biology of Blood and Marrow Transplantation 2012 18, 66-75DOI: (10.1016/j.bbmt.2011.05.010) Copyright © 2012 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 5 Comparison of CMV-specific polyfunctional CD8+ T cells between patients with no antigenemia and those with antigenemia. (A) Represents the number of polyfunctional T cells/L at day 30 in patients with no (n = 5†) and any subsequent antigenemia (n = 7†) at day 30 regardless of stem cell source. (B) Represents the number of CMV-specific CD4 T cells/L (by LDA) at day 30 in patients without (n = 20†) and any subsequent antigenemia (n = 11†). †Includes patients who were eligible for randomization but did not participate in the randomized trial. Bars represent the mean value within each group, and error bars are the standard error of the mean. Biology of Blood and Marrow Transplantation 2012 18, 66-75DOI: (10.1016/j.bbmt.2011.05.010) Copyright © 2012 American Society for Blood and Marrow Transplantation Terms and Conditions

Supplemental Figure S1 Biology of Blood and Marrow Transplantation 2012 18, 66-75DOI: (10.1016/j.bbmt.2011.05.010) Copyright © 2012 American Society for Blood and Marrow Transplantation Terms and Conditions