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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. 2 — Lethal GvHD in fully MHC-matched MCM post-allogeneic HSCT. a Longitudinal white blood cell (WBC), T cell, and platelet absolute counts in recipient macaques 33450 and 33455. Immune conditioning regimens are indicated in top panels (Bu = busulfan, Flu = fludarabine, TBI = total body irradiation, CD3-IT = CD3-immunotoxin). *Fludarabine was administered only to 33450. Colored circumflexes (^) indicate timepoints at which donor-derived cells were detected in whole blood by chimerism assays. Crosses (†) indicate time of necropsy. Normal WBC reference range = 5,600–18,4000/μl; normal platelet reference range = 134,000–564,000/μl. Shaded gray boxes in a, b indicate tacrolimus treatment period. b Longitudinal donor chimerism levels as measured by Illumina sequencing whole blood genomic DNA across SNPs differing between donor and recipient. Graphs display mean ± SEM frequencies of donor-derived cells as measured by two SNPs. c Longitudinal clinical GvHD scoring. Scoring criteria described in Methods section. d Multifocally extensive exfoliative dermatitis of thoracic limb of recipient macaque 33450. e Blinded tissue histopathology GvHD scoring at time of necropsy. Scoring criteria described in Methods section. f Tissue donor chimerism levels at time of necropsy as measured by Illumina sequencing genomic DNA across SNPs differing between donor and recipient. Graphs display mean ± SEM frequencies of donor-derived cells as measured by sequencing two SNPs. To eliminate non-immune cell contamination, colon, jejunum, and liver cell preparations were sorted for CD45+ cells prior to genomic DNA extraction. g Frequencies of Ki67+ T cells in tissues at time of necropsy, as determined by intracellular Ki67 flow cytometric staining. h Mixed lymphocyte reactions assessing levels of CD4+ and CD8+ T cell alloreactivity in recipient macaques 33450 and 33455 post-HSCT (at time of necropsy). 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.