monoclonal anti 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: mBio

Article Title: Metformin as antiviral therapy protects hyperglycemic and diabetic patients

doi: 10.1128/mbio.00634-25

Figure Lengend Snippet: Treatment effectiveness of metformin against lethal SFTSV infection in BKS-db/db mice. ( A ) Survival curves of metformin- or vehicle-treated BKS-db/bd mice ( n = 10 per group) that were pretreated with anti-IFNAR1 IgG antibody and infected with SFTSV (4 × 10 4 PFU per mouse). The Kaplan-Meier method was used to analyze time-to-event data. ( B–E ) Viral loads in serum ( B ), spleen ( C ), liver ( D ), and lung ( E ) from SFTSV-infected SK-db/bd mice with or without (control group) metformin treatment at 5 days post-infection (dpi) ( n = 6 per group). Data were presented as mean ± s.d. The two-sided P values were examined using Student’s t test. ( F ) Representative hematoxylin and eosin (H&E) images from three biologically independent samples of spleen, liver, and lung sections from SFTSV-challenged BKS-db/bd mice with or without metformin treatment at 5 dpi. Scare bar: 500 µm. ( G, H ) Liver ( G ) and spleen samples ( H ) from SFTSV-challenged BKS-db/bd mice with or without metformin treatment at 5 dpi were analyzed by immunoblotting with the indicated antibodies ( n = 3 biologically independent samples).

Article Snippet: To evaluate the antiviral effect of metformin, a lethal mouse model was established using an anti-interferon α/β receptor subunit 1 (IFNAR1) blocking antibody (Bio X Cell, Cat. BE0241) pretreatment ( ).

Techniques: Infection, Control, Western Blot

Journal: Vaccine

Article Title: Immunogenicity and protection of a triple repeat domain III mRNA vaccine against Zika virus.

doi: 10.1016/j.vaccine.2024.126518

Figure Lengend Snippet: Fig. 4. Analysis of long-term immunity by ZIKV-3xEIII against ZIKV. (A) Group design and (B) immunization and challenge schedule of mice. The anti-IFNAR1 blocking antibody was intraperitoneally injected at 1 day before the virus challenge. (C) The levels of ZIKV E-specific antibodies had been measured in serum since boosting. (D) Neutralizing antibodies in serum 40 weeks after boosting by the PRNT. (E) The ZIKV E-specific IgG1 level was measured in serum after challenge. (F) Percentages of body weight of challenged mice were measured every day. (G) Viral titer in various organs of mice were measured by real-time PCR 9 days after virus challenges. (H) Effector memory and (I) effector CD8+ and CD4+ T cells in the spleen and (J) the expression of CD25 and CD69 in CD8+ and CD4+ T cells and (K) The percentages of IFN-γ- and TNF-α-producing CD8+ and CD4+ T cell in the spleen were analyzed using flow cytometry. The isolated splenocytes were stimulated with 500 ng/well of ZIKV E protein or 1 μg/well of peptide mixture. n = 5, Data represent mean ± SD. *p ≤0.05, **p ≤0.01, ***p ≤0.005 and ****p ≤0.001.

Article Snippet: To study the effects of ZIKV challenge in C57BL/6 mice, 2 mg of anti-IFNAR1 blocking antibody (clone MAR1-5A3, BioXCell, Lebanon, NH, USA) was intraperitoneally injected one day before the viral infection.

Techniques: Blocking Assay, Injection, Virus, Real-time Polymerase Chain Reaction, Expressing, Flow Cytometry, Isolation

Journal: Nature

Article Title: Lysine Catabolism Reprograms Tumour Immunity through Histone Crotonylation

doi: 10.1038/s41586-023-06061-0

Figure Lengend Snippet: a, IB analysis of GSC23 after GCDH KD at the indicated time. b, ELISA quantification of IFNα and IFNβ in culture supernatants from GSCs with or without GCDH KD. nd, not detected. c, d, IB analysis of GSC23 treated with IFNβ (5 ng/ml, c) or culture supernatants from GSC23 with or without GCDH KD (d) for indicated time. IFNβ was added every 2 days for the duration of experiments. The supernatants with or without IFNAR blocking antibody were replaced every 2 days. e, IF staining for IFNα, Kcr, Kac, Kglu and GCDH in indicated sections from GSC23-derived intracranial tumours (n = 3 biologically independent mice). Scale bar, 20 μm. f, RT-qPCR analysis of human ISGs in GSC23-derived intracranial tumour tissues (n = 4 biologically independent mice). g, The gating strategy of GSCs in flow cytometric analysis. h, i, Flow cytometry plots (h) and quantification (i, n = 4 biologically independent mice) of SOX2+ CD133+ GSCs in CD147+ human tumour cells as indicated. Representative of two independent experiments in a, c and d. Data are presented from three independent experiments in b. In b, f and i, data are presented as mean ± SEM. One-way ANOVA followed by multiple comparisons with adjusted p values for f and i.

Article Snippet: To block GCDH loss induced activation of IFN signalling, GSCs were preincubated with 10 μg/ml of human IFNAR blocking antibody (cat: MAB4015; R&D Systems) for 2 h in 37 °C, followed with stimulation of the culture supernatants from control or GCDH KD GSCs.

Techniques: Enzyme-linked Immunosorbent Assay, Blocking Assay, Staining, Derivative Assay, Quantitative RT-PCR, Flow Cytometry