1. Supplementary Figure S1
Step 1 (single)
Step 2 (triple)
1x106/kg, single
n = 3
Cohort 1
Cohort 3
1x106/kg, Triple
n = 3
Cohort 4
3x106/kg, Triple
n = 3
Cohort 2
1x107/kg, single
n = 3
1x107/kg, Triple
n = 4
Cohort 5
Cohort 6
3x107/kg, Triple
n = 4
Supplementary Figure S1. Clinical treatment protocol. Step 1 includes single-dose infusion of 1×106 or 1×107 allogeneic NK cells/kg. When each single dose was determined to be safe, step 2, a triple infusion, was started. Arrows indicate the sequential order of administration procedure among cohorts. 2

2. Supplementary Figure S2B
CD16+CD56+
CD3+
CD14+
CD19+ 20 40 60 80
100
% of expression
92.93 ± 2.1%
Viability 20 40 60 80
100
Viability (%) C D D0
D14
100
101
102
103
104
Fold expansion
3:1
1:1
0.3:1 20 40 60 80
100
E:T ratio
% lysis
757.5 ± 223.2
Supplementary Figure S2. Characterization of ex vivo-expanded NK cells. (A) T cell-depleted PBMCs from healthy donors were expanded for 14 days in a GMP-compliant facility. The percentages of CD16+CD56+, CD3+, CD14+ and CD19+ cells were analyzed by flow cytometric analyses (n = 42). (B) The viability of expanded NK cells was evaluated by staining with propidium iodide (n = 16). (C) NK cell numbers were assessed before (D0) and after (D14) NK cell expansion and are expressed as fold expansion (n = 16). (D) Cytotoxicity of expanded NK cells against the K562 cell line was analyzed by flow cytometric cytotoxicity assay with the indicated E:T ratio in triplicate (n = 42). Each point represents mean ± SD. 1

3. Supplementary Figure S3
donor KIR A/A
donor KIR A/B B
Supplementary Figure S3. Analysis of donor KIR genotype and clinical outcome. Donors with KIR genotype A/A (n=7) or A/B (n=10) were reclassified by clinical outcome of recipients (A). Each number of patients with SD (open bar) or PD (solid bar) from donor KIR genotype A/A or KIR genotype A/B is presented (B). SD, stable disease; PD, progressive disease. 3

4. Supplementary Figure S4
Cohort 6-2
Sensitivity
1st
2nd
3rd therapy
HGH
Donor
HLA-DRB1 B
Cohort 6-4
Supplementary Figure S4. Kinetics of donor NK cells in the peripheral blood of recipients. Genomic DNA was extracted from serially-acquired PBMCs of the recipients. The persistence of donor NK cells in the peripheral blood of recipients was determined by allo-HLA-DRB1-specific PCR as described in the Materials and Methods. The sensitivity of nested PCR was analyzed by mixing decreasing amount of DNA from each donor with DNA of a recipient to give final concentrations of 10%, 1%, 0.1%, 0.01% and 0.001% (vol/vol). In Figure A and B, each gel picture shows result of three independent PCR reactions from serially obtained sera of cohort 6-2 (A) and 6-4 (B). The upper band for the HGH gene served as an internal positive control. Lower band indicates donor HLA-DRB1. Each assay was performed three times. 4

5. Supplementary Figure S5
FSC-A F S C - H
98.5 A
5.24 :
CD14&CD19
91.4
CD3
CD56
9.76
4.13
75.2
10.9
CD4
CD8
51.3
0.4
32.2
16.1
NKG2D
99.1
0.286
97.7 NK
CD8+ T
CD4+ T B
Supplementary Figure S5. NKG2D expression on NK and T cells in patients’ peripheral blood. The percentage of NKG2D+ NK cells, CD8 + T cell and CD4 + T cells was analyzed in lympho-gating of patients’ PBMCs by flow cytometry. Representative FACS dot plots and histograms are presented (B). Gating strategy and representative FACS dot plots are presented (A). 5