Journal: Journal for immunotherapy of cancer

Article Title: CD155 blockade enhances allogeneic natural killer cell-mediated antitumor response against osteosarcoma.

doi: 10.1136/jitc-2023-008755

Figure Lengend Snippet: Figure 1 IL-15 expansion of murine NK cells results in activated murine NK. B6 and BALB/c NK cells cultured with IL-15/IL- 15Rα conjugate with additional media and cytokine repletion every 2–3 days are shown. (A) Cell count and (B) fold expansion at days 0, 7, 9, 12, and 14. Percentage of (C) B6 and BALB/c NK cells with marker expression at day 0 and day 12 shown with individual experimental replicates plotted with mean and SEM. (D) Fold change of marker expression MFI at day 12 compared with day 0 shown for B6 NK cells and BALB/c NK cells. Mean with SEM shown for experimental replicates (n=5). Two-sided two-sample t-tests were performed (*p<0.05, **p<0.01, ***p<0.001, ****p>0.0001). IL, interleukin; MFI, median fluorescence intensity; NK, natural killer.

Article Snippet: NK cells were expanded and activated for 14 days using mouse 10 ng/mL IL- 15/ IL- 15Rα conjugate, comprised of recombinant mouse IL- 15 protein (R&D Systems, Minneapolis, Minnesota, USA) and recombinant mouse IL- 15R alpha Fc chimera protein (R&D Systems), in Roswell Park Memorial Institute medium (Corning Life Sciences, Durham, North Carolina, USA) with 10% fetal bovine serum (GeminiBio, Sacramento, California, USA) with 50 IU/mL penicillin/ streptomycin (Lonza), 0.11 mM 2- betamercaptoethanol (Gibco, Carlsbad, California, USA), 1X MEM nonessential amino acids (Corning), 25 mM HEPES buffer P ro tected b y co p yrig h t, in clu d in g fo r u ses related to text an d d ata m in in g , A I train in g , an d sim ilar tech n o lo g ies. . b y g u est o n A p ril 20, 2025 h ttp ://jitc.b m j.co m / D o w n lo ad ed fro m 15 A p ril 2025.

Techniques: Cell Culture, Cell Counting, Marker, Expressing, Fluorescence

Journal: Journal for immunotherapy of cancer

Article Title: CD155 blockade enhances allogeneic natural killer cell-mediated antitumor response against osteosarcoma.

doi: 10.1136/jitc-2023-008755

Figure Lengend Snippet: Figure 4 AlloBMT and single or multiple dose CD155 blockade with alloNK effectively treat relapsed OS pulmonary metastases. (A) Schematic showing alloBMT followed by K7M2 OS inoculation and single dose or multiple doses of alloNK infusion with CD155 axis blocking antibodies. (B) Serum levels of mouse IL-15/IL-15R⍺ pre- and post-BMT and single dose alloNK and CD155 axis blockade treatment at time points post alloBMT (n=6–15 mice per time point) (C) EFS for mice treated with alloBMT and single dose alloNK with isotype control or anti-CD155 blockade are shown at time points post alloBMT (n=8 or 10 mice per group). (D) Clinical GVHD scores for mice treated with alloBMT and single dose alloNK with isotype control or anti- CD155 blockade are shown (n=8 or 10 mice per group). (E) EFS for mice treated with alloBMT and multiple doses of alloNK with isotype control or anti-CD155 blockade are shown at time points post alloBMT (n=3 mice per group). (F) Clinical GVHD scores for mice treated with alloBMT and multiple doses of alloNK with isotype control or anti-CD155 blockade are shown (n=3 mice per group). Bars and points with error bars depict mean with SEM. Log-rank test performed and student’s t-tests were performed (*p<0.05). alloBMT, allogeneic bone marrow transplant; alloNK, allogeneic natural killer; BMT, bone marrow transplant; EFS, event-free survival; GVHD, graft-versus-host disease; IL, interleukin; NK, natural killer; OS, osteosarcoma.

Article Snippet: NK cells were expanded and activated for 14 days using mouse 10 ng/mL IL- 15/ IL- 15Rα conjugate, comprised of recombinant mouse IL- 15 protein (R&D Systems, Minneapolis, Minnesota, USA) and recombinant mouse IL- 15R alpha Fc chimera protein (R&D Systems), in Roswell Park Memorial Institute medium (Corning Life Sciences, Durham, North Carolina, USA) with 10% fetal bovine serum (GeminiBio, Sacramento, California, USA) with 50 IU/mL penicillin/ streptomycin (Lonza), 0.11 mM 2- betamercaptoethanol (Gibco, Carlsbad, California, USA), 1X MEM nonessential amino acids (Corning), 25 mM HEPES buffer P ro tected b y co p yrig h t, in clu d in g fo r u ses related to text an d d ata m in in g , A I train in g , an d sim ilar tech n o lo g ies. . b y g u est o n A p ril 20, 2025 h ttp ://jitc.b m j.co m / D o w n lo ad ed fro m 15 A p ril 2025.

Techniques: Blocking Assay, Control

Journal: Journal for immunotherapy of cancer

Article Title: CD155 blockade enhances allogeneic natural killer cell-mediated antitumor response against osteosarcoma.

doi: 10.1136/jitc-2023-008755

Figure Lengend Snippet: Figure 5 CD155 blockade with alloNK shows limited efficacy against established OS disease after alloBMT. (A) Schematic showing K7M2 OS inoculation, alloBMT, and alloNK infusion with CD155 axis blocking antibodies. (B) Blood serum levels of mouse IL-15/IL-15R⍺ pre-BMT and post-BMT and alloNK and CD155 axis blockade treatment (n=7–8 mice per time point) (C) EFS for mice treated with alloBMT and alloNK with isotype control, anti-DNAM-1 blockade, anti-CD155 blockade, or anti- DNAM and anti-CD155 blockade are shown (n=7–8 mice per group). (D) Clinical GVHD scores for mice treated with alloBMT and alloNK with isotype control, anti-DNAM-1 blockade, anti-CD155 blockade, or anti-DNAM and anti-CD155 blockade are shown (n=7–8 mice per group). (E) Tumor burden measured by IVIS shown as total flux (photons/s) for mice treated with alloBMT and alloNK with isotype control or anti-CD155 blockade (n=6 per group). (F) Recovery of infused alloNK expressing CD45.1 from spleens harvested from mice treated with alloBMT and alloNK with isotype control or anti-CD155 blockade (n=3 mice per time point). Bars and points with error bars depict mean with SEM. alloBMT, allogeneic bone marrow transplant; alloNK, allogeneic natural killer; BMT, bone marrow transplant; EFS, event-free survival; GVHD, graft-versus-host disease; IL, interleukin; IVIS, in vivo imaging system; NK, natural killer; OS, osteosarcoma.

Article Snippet: NK cells were expanded and activated for 14 days using mouse 10 ng/mL IL- 15/ IL- 15Rα conjugate, comprised of recombinant mouse IL- 15 protein (R&D Systems, Minneapolis, Minnesota, USA) and recombinant mouse IL- 15R alpha Fc chimera protein (R&D Systems), in Roswell Park Memorial Institute medium (Corning Life Sciences, Durham, North Carolina, USA) with 10% fetal bovine serum (GeminiBio, Sacramento, California, USA) with 50 IU/mL penicillin/ streptomycin (Lonza), 0.11 mM 2- betamercaptoethanol (Gibco, Carlsbad, California, USA), 1X MEM nonessential amino acids (Corning), 25 mM HEPES buffer P ro tected b y co p yrig h t, in clu d in g fo r u ses related to text an d d ata m in in g , A I train in g , an d sim ilar tech n o lo g ies. . b y g u est o n A p ril 20, 2025 h ttp ://jitc.b m j.co m / D o w n lo ad ed fro m 15 A p ril 2025.

Techniques: Blocking Assay, Control, Expressing, In Vivo Imaging