Journal: Cancer Immunology, Immunotherapy

Article Title: IL-15 superagonist N-803 improves IFNγ production and killing of leukemia and ovarian cancer cells by CD34 + progenitor-derived NK cells

doi: 10.1007/s00262-020-02749-8

Figure Lengend Snippet: N-803 enhances HPC-NK cell proliferation, IFNγ production, and leukemia killing. a–b ( a ) Percentage of proliferating HPC-NK cells based on proliferation dye eFluor450 staining ( n = 3–4), ( b ) number of HPC-NK cells based on CD56 antibody staining ( n = 4–5) 6 days after incubation with no cytokine (white), rhIL-15 (grey) or N-803 (black). c Percentage of IFNγ + HPC-NK cells 4 h after incubation with K562 or THP-1, with (black) or without (white) 1 nM N-803 combined for NK only ( n = 6), K562 ( n = 6) or THP-1 ( n = 4). d IFNγ concentration (pg/ml) after overnight co-culture of HPC-NK cells and K562 ( n = 3) or THP-1 ( n = 4) with or without 1 nM N-803 (without, white; with N-803, black, ND = not detectable). e Geometric mean fluorescence intensity (MFI) of ICAM-1 expression after overnight culture of K562 ( n = 3) or THP-1 ( n = 4, white), and addition of N-803 (light grey), HPC-NK cells (dark grey) or both (black). f – g Percentage of ( f ) K562 ( n = 3) or ( g ) THP-1 ( n = 4) killing after overnight co-culture with HPC-NK cells and 0 (grey) or 1 nM N-803 (black). h IFNγ concentration (pg/ml) after 48 h co-culture of HPC-NK cells and primary AML (pAML) cells ( n = 5) with 1 nM N-803 (black) or without cytokine (white). i Delta median fluorescence intensity (ΔMFI) of ICAM-1 expression after 48 h culture of pAML cells ( n = 4, white), addition of N-803 (lightest grey), HPC-NK cells or both (different shades of grey/black) compared to a backbone sample for each condition. j Percentage of pAML cell killing ( n = 5) after 48 h co-culture with HPC-NK cells and 0 (white) or 1 nM N-803 (black). Graphs show mean ± SEM for a–g , i – j. One-way ANOVA with Bonferroni correction was used (repeated measures for f – g , i – j ) to test for statistical significance

Article Snippet: NK cells were labeled with eFluor450 (eBioScience, 65-0842-85) and cultured in NK MACS/10% HS with/without rhIL-15 or N-803 (ImmunityBio).

Techniques: Staining, Incubation, Concentration Assay, Co-Culture Assay, Fluorescence, Expressing

Journal: Cancer Immunology, Immunotherapy

Article Title: IL-15 superagonist N-803 improves IFNγ production and killing of leukemia and ovarian cancer cells by CD34 + progenitor-derived NK cells

doi: 10.1007/s00262-020-02749-8

Figure Lengend Snippet: HPC-NK cells combined with N-803 and nanogam show anti-tumor effects in mice bearing human OC. ( a ) Experiment 1: number of HPC-NK cells per ml in the peritoneal wash of SKOV-3 bearing NSG mice (0.2 million tumor cells injected 4 days before HPC-NK cell treatment) 15/16 days after treatment with HPC-NK cells in combination with phosphate-buffered saline (PBS), 2.5 µg rhIL-15, or 50 or 200 µg/kg N-803 (6 mice per group). (b ) Experiment 2: number of HPC-NK cells per ml in the peritoneal wash of SKOV-3 bearing NSG mice (0.2 million tumor cells injected 4 days before HPC-NK cell treatment) 14/15 days after treatment with HPC-NK cells in combination with 50 µg/kg N-803, 2.5 µg rhIL-15, 50 µg/kg N-803 + nanogam, 50 µg/kg N-803 + 2.25 Gy irradiation, or 2.5 µg rhIL-15 + 2.25 Gy irradiation (6 mice per group). (c ) Experiment 3: radiance (photons/second/cm 2 /steradian) of tumors in SKOV-3 bearing NSG mice (0.2 million tumor cells injected 4 days before the first HPC-NK cell infusion) treated with nanogam + PBS (white, control), nanogam + 2 HPC-NK cell infusions + 2.5 µg rhIL-15 (grey) or nanogam + 2 HPC-NK cell infusions + 50 µg/kg N-803 (black) over time (7 mice per group). (d ) BLI images from ( c ) acquired over time. One-way ANOVA with Bonferroni correction was used for a – b (after log transformation for a and b ), and a repeated-measures two-way ANOVA with Bonferroni correction for c to test for statistical significance

Article Snippet: NK cells were labeled with eFluor450 (eBioScience, 65-0842-85) and cultured in NK MACS/10% HS with/without rhIL-15 or N-803 (ImmunityBio).

Techniques: Injection, Irradiation, Transformation Assay