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. 2017 Nov 10;8:1418. doi: 10.1038/s41467-017-01631-z

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — Stable, multi-lineage donor chimerism in MHC-matched MCM post-allogeneic HSCT. a Longitudinal white blood cell (WBC), T cell, and platelet absolute counts in recipient macaques 33454 and 34666. Immune conditioning regimens are indicated in top panels (Bu = busulfan, TBI = total body irradiation, CD3-IT = CD3-immunotoxin). Normal WBC reference range = 5,600–184,000/μl; normal platelet reference range = 134,000–564,000/μl. Shaded gray boxes in a, b indicate tacrolimus treatment period. b, c Donor chimerism levels as measured by Illumina sequencing genomic DNA across SNPs differing between donor and recipient. Specific populations were isolated by flow cytometric cell sorting of ACK-treated whole blood prior to genomic DNA extraction. Longitudinal whole blood (WB), granulocyte, and T cell donor chimerism levels shown in b. Timepoints of donor lymphocyte infusions (DLIs) into 33454 are indicated with arrows in b (bottom panel); DLI #1 = 1 × 10⁷ CD3+ cells/kg, DLI #2 = 5 × 10⁷ CD3+ cells/kg, DLI #3 = 5 × 10⁷ CD3+ cells/kg. Comprehensive immune cell subset donor chimerism levels in whole blood (top) and lymph node (bottom) shown in c, measured 428 days (33454) or 193 days (34666) post-HSCT. N.D. = not determined due to insufficient cell numbers. d Mixed lymphocyte reactions assessing levels of CD4+ and CD8+ T cell alloreactivity in recipient macaques 33454 and 34666 post-HSCT (354 and 298 days post-HSCT, respectively). Plots display proliferation of CFSE-labeled recipient T cells in response to irradiated donor or pre-HSCT recipient cells, as measured by frequency of CFSE-lo cells. Plots are gated on live, CD3+ singlets, and either CD4+ or CD8+ cells. Recipient cells cultured alone (no stim) or with SEB served as negative and positive controls, respectively.