Quantitative Assessment of T Cell Clonotypes in Human Acute Graft-versus-Host Disease Tissues Daisuke Koyama, Makoto Murata, Ryo Hanajiri, Tomohiro Akashi,

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Quantitative Assessment of T Cell Clonotypes in Human Acute Graft-versus-Host Disease Tissues Daisuke Koyama, Makoto Murata, Ryo Hanajiri, Tomohiro Akashi, Shingo Okuno, Sonoko Kamoshita, Jakrawadee Julamanee, Erina Takagi, Kotaro Miyao, Reona Sakemura, Tatsunori Goto, Seitaro Terakura, Tetsuya Nishida, Hitoshi Kiyoi Biology of Blood and Marrow Transplantation Volume 25, Issue 3, Pages 417-423 (March 2019) DOI: 10.1016/j.bbmt.2018.10.012 Copyright © 2018 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure1 Comparison of clonotype distributions among GVHD tissues and peripheral blood (PB). (A) Clonotype distribution plots of T cells in the skin and concurrent PB obtained from patient 1. Each dot represents a distinct CDR3 amino acid sequence. Arrows indicate the most frequent and second most frequent clonotypes in the skin. (B) Diversity of the TCR repertoire (1/Ds) of T cells in the GVHD tissues (n = 12), concurrent PB (n = 8), and normal PB from healthy volunteers (n = 3). *P < .05; ****P < .0001. Biology of Blood and Marrow Transplantation 2019 25, 417-423DOI: (10.1016/j.bbmt.2018.10.012) Copyright © 2018 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure2 Allogeneic responses of isolated T cell clones. Cytotoxicities of T cell clones 1 and 2 against patient PBMCs (solid squares) and donor EBV-LCLs (open squares) were determined by a chromium release assay. Specific lysis is shown as the mean of triplicate cultures at various E:T ratios. Biology of Blood and Marrow Transplantation 2019 25, 417-423DOI: (10.1016/j.bbmt.2018.10.012) Copyright © 2018 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure3 Clonotype distribution plots of T cells in the skin, stomach, colon, and concurrent PB obtained from patient 4. Each dot represents a distinct CDR3 amino acid sequence. Arrows indicate the most frequent and second most frequent clonotypes in the skin and stomach and the most frequent clonotype in the colon. Biology of Blood and Marrow Transplantation 2019 25, 417-423DOI: (10.1016/j.bbmt.2018.10.012) Copyright © 2018 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure4 Comparison of frequencies of clonotypes in each GVHD tissue and in PB. The dot plot shows clonotype distributions in T cells that accounted for ≥5% in the skin (n = 41), stomach (n = 19), or colon (n = 7) and their frequencies in the concurrent PB of the corresponding patient. The number in the upper right corner is the Spearman correlation coefficient, ρ. Biology of Blood and Marrow Transplantation 2019 25, 417-423DOI: (10.1016/j.bbmt.2018.10.012) Copyright © 2018 American Society for Blood and Marrow Transplantation Terms and Conditions