ifn i receptor subunit 1 ifnar1 blocking antibody  (Bio X Cell)

Journal: Cancer Research

Article Title: Targeting Intracellular Innate RNA-Sensing Systems Overcomes Resistance to CAR T-cell Therapy in Solid Tumors

doi: 10.1158/0008-5472.CAN-24-3425

Figure Lengend Snippet: Tumor-intrinsic MAVS signaling induces susceptibility to CAR T-cell killing via soluble factors and auto-/paracrine IFN signaling that can spread to bystander tumor cells. A, B16-EpCAM cells were pretreated with anti-IFNaR1 blocking antibody before 3pRNA transfection and subsequent coculture with anti-EpCAM CAR or UTD T cells. Apoptosis induction in tumor cells after 24 hours was determined by flow cytometry. Data are mean ± SEM of n = 3 replicates. B, WT or Mavs − / − B16-EpCAM cells were either directly transfected with 3pRNA or exposed to conditioned culture medium from 3pRNA-activated WT B16-EpCAM cells. Relative expression of Puma , Bid , and Ifnb1 in bystander B16-EpCAM cells was determined by RT-PCR. Data were normalized to unstimulated (unstim.) B16 cells and are mean ± SEM of n = 3 replicates. C, WT or Mavs − / − B16-EpCAM cells were exposed to anti-EpCAM CAR or UTD T cells in the presence of conditioned culture medium from 3pRNA-activated WT B16-EpCAM cells. Apoptosis induction by flow cytometry is presented as mean ± SEM of n = 6 replicates. All data are representative of or pooled from at least two independent experiments.

Article Snippet: For some experiments, tumor cells or CAR T cells were incubated for a minimum of 2 hours at 37°C with 20 μg/mL monoclonal anti-IFNaR1 blocking antibody (clone MAR1-5A3, RRID: AB_2687723, Bio X Cell).

Techniques: Blocking Assay, Transfection, Flow Cytometry, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Cancer Research

Article Title: Targeting Intracellular Innate RNA-Sensing Systems Overcomes Resistance to CAR T-cell Therapy in Solid Tumors

doi: 10.1158/0008-5472.CAN-24-3425

Figure Lengend Snippet: Therapeutically activated tumor-intrinsic RIG-I/MAVS signaling imprints a cytolytic phenotype on CAR T cells. A and B, B16-EpCAM tumor cells were transfected with 3pRNA prior to exposure to anti-EpCAM CAR T cells or UTD control T cells. Expression of CD69, IFNγ, and granzyme B ( A ) and Fas ligand (FasL; B ) on T cells was measured by flow cytometry 24 hours after coculture. Data are mean ± SEM of n = 6 replicates. C, Anti-EpCAM CAR or UTD T cells were cocultured with 3pRNA-treated WT or Mavs − / − B16-EpCAM for 24 hours before rechallenge with steady-state WT B16-EpCAM cells. Apoptosis induction in initial and rechallenge B16 cells was determined by flow cytometry. All data are mean ± SEM of n = 6 replicates. D, Anti-EpCAM CAR T cells were exposed to conditioned culture medium from RIG-I–treated B16-EpCAM cells. Expression of CD25, PD-1, CD69, and granzyme B on CAR T cells is presented as mean ± SEM of n = 3 replicates. E, Apoptosis induction in 3pRNA-treated B16-EpCAM cells 24 hours after coculture with WT or Ifnar1 − / − anti-EpCAM CAR or UTD T cells. Data are mean ± SEM of n = 4 replicates. F, Heatmap showing the relative release of cytokines and chemokines from B16-EpCAM, Panc02-EpCAM, and MG846-MSLN cells in response to RIG-I activation. All data are representative of or pooled from at least two independent experiments. Unstim., unstimulated. C, Created in BioRender. Soliman, N. (2025) https://BioRender.com/l0n2a28 .

Article Snippet: For some experiments, tumor cells or CAR T cells were incubated for a minimum of 2 hours at 37°C with 20 μg/mL monoclonal anti-IFNaR1 blocking antibody (clone MAR1-5A3, RRID: AB_2687723, Bio X Cell).

Techniques: Transfection, Control, Expressing, Flow Cytometry, Activation Assay

Journal: Cells

Article Title: TLR9 Monotherapy in Immune-Competent Mice Suppresses Orthotopic Prostate Tumor Development

doi: 10.3390/cells13010097

Figure Lengend Snippet: RM1 cells are sensitive to the growth inhibitory effects of IFN-β secreted by CPG-1668-stimulated macrophages. ( A ) RM1 cells were treated with various concentrations of recombinant cytokines (IFN-β, IFN-γ, or TNFα) for 24, 48, or 72 h; then, cell viability was determined by MTS absorbance. ( B ) Cell death (measured by AnnexinV-FITC and propidium iodide (PI) staining) or ( C ) proliferation (measured by the total number of live cells) was also assessed after incubation with IFN-β at the same time points. ( D ) The colony-forming potential of RM1 cells was determined after 24 h incubation with recombinant cytokines. ( E ) The clonogenicity of cells following cotreatment with CPG-1668 (10 µg/mL) and IFN-γ (100 ng/mL) or IFN-β (100 ng/mL) was also determined. ( F ) The amount of IFN-β secreted into the culture media of BMDMs grown in various concentrations of CPG-1668 was determined by ELISA. ( G ) RM1 cells were grown in BMDM-conditioned media (CM) for 24 h; then, the colony-forming potential was determined. ( H ) Some cells were pretreated with 50 µg/mL IFNAR1 blocking monoclonal antibody for 1 h before the addition of BMDM-derived CM or recombinant IFN-β (1 ng/mL). The colony-forming potential after 24 h or cell proliferation after 48 h was determined. Data represent n = 3–4 per group and expressed as mean ± SEM. Statistical analysis was conducted using ( D – G ) ordinary one-way ANOVA followed by Dunnett’s post hoc test for multiple comparisons or ( A – C , H ) two-way ANOVA followed by Sidak’s post hoc test for multiple comparisons (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: In some experiments, cells were pretreated with 50 μg/mL antimouse IFNAR1 blocking monoclonal antibody (BioXcell, Lebanon, NH, USA) prior to treatment.

Techniques: Recombinant, Staining, Incubation, Enzyme-linked Immunosorbent Assay, Blocking Assay, Derivative Assay

Journal: Transfusion

Article Title: Type 1 IFN signaling critically regulates influenza-induced alloimmunization to transfused KEL RBCs in a murine model.

doi: 10.1111/trf.15482

Figure Lengend Snippet: Fig. 4. Type 1 IFN receptor blockade prevents influenza-induced alloimmune responses to transfused K1 RBCs. (A) Wild-type C57BL/6 mice were treated with IFNAR1 blocking antibody (MAR1-5A3) or an isotype matched IgG1 control 24 hours before influenza infection,

Article Snippet: For blocking experiments, mice were intraperitoneally injected with 600 μg anti-mouse IFNAR1 blocking antibodies (clone: MAR1-5A3, BioXCell), isotype control IgG1 (MOPC-21, BioXCell) or phosphate-buffered saline 24 hours before PR8 infection, 24 hours before transfusion, and 3 days after transfusion.

Techniques: Blocking Assay, Control, Infection