Addition of 10-Day Decitabine to Fludarabine/Total Body Irradiation Conditioning is Feasible and Induces Tumor-Associated Antigen-Specific T Cell Responses  Marjan Cruijsen, Willemijn Hobo, Walter J.F.M. van der Velden, Manita E.J. Bremmers, Rob Woestenenk, Brigitte Bär, J.H. Frederik Falkenburg, Michel Kester, Nicolaas P.M. Schaap, Joop Jansen, Nicole N.M. Blijlevens, Harry Dolstra, Gerwin Huls  Biology of Blood and Marrow Transplantation  Volume 22, Issue 6, Pages 1000-1008 (June 2016) DOI: 10.1016/j.bbmt.2016.02.003 Copyright © 2016 The American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 1 The Dec/Flu/TBI conditioning regimen. In mismatched unrelated transplants, ATG 2 mg/kg/day was added on days -8, -7, -6, and -5. Biology of Blood and Marrow Transplantation 2016 22, 1000-1008DOI: (10.1016/j.bbmt.2016.02.003) Copyright © 2016 The American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 2 OS after Dec/Flu/TBI conditioning at a median follow-up of 443 days post–allogeneic HCT. Biology of Blood and Marrow Transplantation 2016 22, 1000-1008DOI: (10.1016/j.bbmt.2016.02.003) Copyright © 2016 The American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 3 Detection of TAA-reactive CD8+ T cell responses in peripheral blood post–allogeneic HCT. Patient PBMCs collected post-allografting were analyzed for the presence of TAA-specific CD8+ T cell responses using combinatorial encoded tetramer-based flow cytometry. Patients were classified with a TAA-specific T cell response when double tetramer-positive CD8+ T cells were found directly after thawing or after 1 week of peptide stimulation. (A) Data for all patients. Color-coding is as follows: dark green indicates a specific T cell response ≥0.01%; light green, a T cell response ≥0.001 to <0.01%; gray, no T cell response detected or not a relevant HLA type. (B) TAA tetramer staining for 4 different patients. Black dots represent tetramer-positive CD8+ T cells within the total CD8+ T cell population (gray). Text in plots indicates the day of analysis on which TAA-reactive T cells were detected. Survival (in weeks) and time point of tetramer analysis (in months) post-allografting are indicated. Pt, patient; y, yes; n, no. *Indicates evaluation of the TAA-specific CD8+ T cell response at another time point post allografting. +Indicates patients with a positive response in this second sample: patient 7 (3 months post-HCT: 0.008% at day 7), patient 15 (6 months post-HCT: MAGE-A3 FLW, 0.012%; MAGE-A2 YLQ, 0.020% at day 0), patient 16 (6 months post-HCT: PRAME, 0.005%; MAGE-A2 YLQ, 0.014%, both at day 7), and patient 18 (8 months post-HCT: 0.046% at day 7). This Figure is available in color online at www.bbmt.org. Biology of Blood and Marrow Transplantation 2016 22, 1000-1008DOI: (10.1016/j.bbmt.2016.02.003) Copyright © 2016 The American Society for Blood and Marrow Transplantation Terms and Conditions