mouse cd137  (R&D Systems)

Journal: iScience

Article Title: Tumor targeted 4-1BB agonist antibody-albumin fusions with high affinity to FcRn induce anti-tumor immunity without toxicity

doi: 10.1016/j.isci.2022.104958

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Article Snippet: Recombinant mouse 4-1BB/CD137 Fc chimera , SinoBiological , Cat#50811-M02H.

Techniques: Recombinant, Cell Isolation, Enzyme-linked Immunosorbent Assay, Luciferase, Expressing, Software

Journal: Cancer Immunology, Immunotherapy

Article Title: Anti-tumor effects of RTX-240: an engineered red blood cell expressing 4-1BB ligand and interleukin-15

doi: 10.1007/s00262-021-03001-7

Figure Lengend Snippet: 4-1BBL and IL-15TP on RTX-240 stimulate receptor-specific activation in reporter cells. a Representative flow cytometry plots of RTX-240 stained for 4-1BBL and IL-15TP. b Fold change in NFkB activation in 4-1BB/NFkB reporter HEK293 cells incubated with engineered RBCs or 4-1BB agonistic antibody + cross-linking antibody (10 nM and 1 nM agonist with 25 nM and 2.5 nM cross-linker, respectively) compared with media alone. c Fold change in JAK/STAT activation by IL-15 in reporter HEK-Blue-IL-2 cells incubated with engineered RBCs or recombinant (r) IL-15 (threefold dilutions from 100 pg/mL), compared with media alone. Bars indicate SD of 2–3 technical replicates. IL-15TP, trans-presented interleukin 15; RBC, red blood cell; SD, standard deviation

Article Snippet: For mRBC, 5 × 10 6 mRBCs were incubated with 1 μg of recombinant mouse 4-1BB human Fc chimera protein (R&D Systems™, Inc.) for 30 min.

Techniques: Activation Assay, Flow Cytometry, Staining, Incubation, Recombinant, Standard Deviation

Journal: Cancer Immunology, Immunotherapy

Article Title: Anti-tumor effects of RTX-240: an engineered red blood cell expressing 4-1BB ligand and interleukin-15

doi: 10.1007/s00262-021-03001-7

Figure Lengend Snippet: Direct activation of CD8 + T cells and NK cells in vitro. ( a i) PBMCs were labeled with CTFR and incubated with the indicated treatments for 8 days, then analyzed by flow cytometry for: a memory CD8 + T cell numbers; b memory CD8 + T cell proliferation (percentage of cells that went through at least one division); c CD8 + effector memory differentiation; d NK cell numbers; e NK cell proliferation (percentage of cells that went through at least one division); f – i expression of the following molecules on NK cells: f TRAIL, g NKp44, h Granzyme B and i 4-1BB. j Purified NK cells were incubated in vitro overnight with the indicated treatments, then incubated with labeled K562 target cells for 4 h. Target cell killing was measured by flow cytometry. Bars indicate a – i SD of 3 biological replicates j or SD of 3 technical replicates. Flow plots are representative data from one donor. CTFR, CellTrace Far Red dye; GzmB, Granzyme B; IL-15TP, trans-presented interleukin-15; NK, natural killer; PBMC, peripheral blood mononuclear cell; rIL-15, recombinant IL-15; SD, standard deviation

Article Snippet: For mRBC, 5 × 10 6 mRBCs were incubated with 1 μg of recombinant mouse 4-1BB human Fc chimera protein (R&D Systems™, Inc.) for 30 min.

Techniques: Activation Assay, In Vitro, Labeling, Incubation, Flow Cytometry, Expressing, Purification, Recombinant, Standard Deviation

Journal: Cancer Immunology, Immunotherapy

Article Title: Anti-tumor effects of RTX-240: an engineered red blood cell expressing 4-1BB ligand and interleukin-15

doi: 10.1007/s00262-021-03001-7

Figure Lengend Snippet: mRBC-240 is tolerated in mice. a Quantification of liver macrophages (F4/80 +). b Representative images of IHC staining of macrophages (F4/80) from selected mice. c Quantification of liver-infiltrating CD8 + T cells and d liver-infiltrating cytotoxic CD8 + T cells (CD8 + /Eomes + /KLRG1 +). Quantification was performed on liver samples of C57BL/6 wild-type mice by flow cytometry on day 18 following 4 doses of either mRBC-240 (1 × 10 9 , 3 × 10 8 , or 1 × 10 8 ), mRBC-CTRL, 4-1BB agonistic antibody (10 mg/kg or 2.5 mg/kg) ( n = 8 mice/group) or PBS. e ALT liver enzyme levels (U/L) in serum on day 18. f Inflammation scoring performed on H&E stained liver sections. g Representative images of H&E staining of liver sections from selected mice. All comparisons were analyzed by a one-way ANOVA and compared with control groups and showing as * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ALT, alanine aminotransferase; IHC, immunohistochemistry; H&E, hematoxylin and eosin; mRBC, mouse red blood cell; PBS, phosphate-buffered saline

Article Snippet: For mRBC, 5 × 10 6 mRBCs were incubated with 1 μg of recombinant mouse 4-1BB human Fc chimera protein (R&D Systems™, Inc.) for 30 min.

Techniques: Immunohistochemistry, Flow Cytometry, Staining, Saline