Bone marrow transplantation for MHC class I deficiency corrects T-cell immunity but dissociates natural killer cell repertoire formation from function

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Bone marrow transplantation for MHC class I deficiency corrects T-cell immunity but dissociates natural killer cell repertoire formation from function Yifang Gao, PhD, Peter D. Arkwright, FRCPCH, DPhil, Rachel Carter, MSc, Angelica Cazaly, PhD, Rebecca J. Harrison, BSc, Alexandra Mant, PhD, Andrew J. Cant, FRCP, FRCPCH, MD, Stephan Gadola, FRCP, PhD, Tim J. Elliott, PhD, Salim I. Khakoo, FRCP, MD, Anthony P. Williams, MRCP, FRCPath, PhD Journal of Allergy and Clinical Immunology Volume 138, Issue 6, Pages 1733-1736.e2 (December 2016) DOI: 10.1016/j.jaci.2016.06.029 Copyright © 2016 The Authors Terms and Conditions

Fig 1 Clinical presentation of the patient showing pretransplant necrotic ulcer of the right elbow (A) and posttransplant healed ulcer of the right elbow at 12 months (B). C, Flow cytometry of MHC class I expression on PBMCs taken pretransplantation and posttransplantation, compared with that of the father (HLA-A2/24; HLA-B35/61; HLA-Cw4/Cw15). Isotype control (filled histograms) and specific anti-HLA antibodies (black unfilled histograms) are shown together with the MFI of MHC class I expression. D, TAP expression by intracellular flow cytometry of patient BLCL pretransplantation and posttransplantation compared with that of the father, mother, and healthy control. Cells were stained for TAP1 (TAP1.28, unfilled histograms black line), TAP2 (TAP2.17, unfilled histograms gray line), or relevant isotype control (filled histograms). Histograms were gated on live lymphocytes. The MFI of the TAP1 and TAP2 expression is shown. BLCL, EBV transformed lymphoblastoid B cell line; MFI, mean fluorescence intensity. Journal of Allergy and Clinical Immunology 2016 138, 1733-1736.e2DOI: (10.1016/j.jaci.2016.06.029) Copyright © 2016 The Authors Terms and Conditions

Fig 2 A, Flow cytometric analysis of T-cell receptor Vβ family expression on pretransplant and 12-month posttransplant T cells (left panel) and on the CD4 or CD8 T-cell subsets posttransplant (right panel). The frequencies of the individual Vβ family within each subpopulation are shown as a percentage of that particular subset. B, Comparison of NK-cell function pretransplant and posttransplant (18 months) with 3 healthy controls. Shown are the percentages of CD3−veCD56+veNK cells expressing either IFN-γ by intracytoplasmic cytokine staining or degranulating (CD107a expression) in response to IL-15, K562 or IL-15 and K562, as determined by flow cytometry. Journal of Allergy and Clinical Immunology 2016 138, 1733-1736.e2DOI: (10.1016/j.jaci.2016.06.029) Copyright © 2016 The Authors Terms and Conditions

Fig E1 A, RT-PCR of the different exons of TAP1 and TAP2 showing absence of TAP1 genes beyond exon 3 and complete absence of TAP2. RNA was extracted from BLCL of the patient, TAP positive (T0) and TAP-negative (T2) control lines. Tapasin (*) was amplified as a loading control. B, PCR from genomic DNA showing the presence of TAP1 up to exon 3 and the absence of TAP2 in the same cell lines as in Fig E1, A. Journal of Allergy and Clinical Immunology 2016 138, 1733-1736.e2DOI: (10.1016/j.jaci.2016.06.029) Copyright © 2016 The Authors Terms and Conditions