Supplementary Data from Complete Remission with Reduction of High-Risk Clones following Haploidentical NK-Cell Therapy against MDS and AML

Abstract

<p>Supplementary Figures and Tables Supplemental Figure 1. Persistence of donor NK cells in vivo. (A) Frequency of patients with detectable donor NK cells in peripheral blood (PB) at day (d)7 and/or 14 after NK cell infusion in nonresponders (NR) and responders (R). (B) Identification of CD56+ donor NK cells based on expression of donor-unique HLA class I molecules (HLA-A2 for P3 and HLA-11 for P11). Shown are donor NK cells in the cell product prior to infusion (upper part) and in the recipient (lower part) day 21 (P3) or day 14 (P11) after NK cell infusion. Supplemental Figure 2. NK cell product repertoires. (A) Representative staining for CD56dim NK cell marker expression. Summary of results for (B) geometrical mean fluorescence intensity (GeoMFI) of perforin, Granzyme B, CXCR4, NKG2D and DNAM expression for CD56dim NK cells. (C) Frequencies of CD56dim NK cells responding with degranulation (CD107a) and IFN-γ production following coculture with K562 target cells. (D) NK cell differentiation stages as defined by CD57, KIR and NKG2A expression. (E) Frequencies of CD56dim NK cells expressing zero, one or multiple KIRs. Non-responders (NR; filled circles, n=3-5) and responders (R; open squares, n=5). Supplemental Figure 3. Clonal evolution in relapse patients. Visualization of presence and allelic burden of clones present in bone marrow of (A) P5 and (B) P14, pre and post NK cell treatment and at relapse. Supplemental Figure 4. Recipient NK cell responses. (A) Separation of donor (Do) and recipient NK cells based on donor unique HLA class I molecules. Shown are representative examples for HLA-A2, HLA-11, HLA-B7 and HLA-B13 with donor NK cells in donor product (gated upper part) and recipient NK cells (gated lower part) at day 14/21 post NK cell infusion. (B) Representative staining of CD69 and Ki-67 on CD3- CD56dim NK cells shown for donor (HLA+) cells and patient (HLA) cells from a non-responder (NR) and a responder (R). (C-D) Summary of results for frequencies of Ki-67+ and CD69+ CD56dim NK cells for all evaluable patients. (Do, filled triangles, n=9), (NR, filled circles, n=4) and (R, open squares, n=3). Supplemental Figure 5. Activation of Treg post NK cell infusion. (A) Representative staining for expression of Ki-67, PD-1 and CTLA-4 for CD3+CD4+CD127- FoxP3+ Treg pre and d28 post NK cell infusion. (B-E) Treg marker expression for non-responder (NR; filled circles, n=6-7) and responders (R; open squares, n=5). Supplemental Tables 1-3</p>