anti ifnar1 monoclonal antibody  (Bio X Cell)

Journal: NPJ Vaccines

Article Title: Durable humoral immunity and long-term protection induced by a Crimean-Congo hemorrhagic fever virus replicon particle vaccine in mice

doi: 10.1038/s41541-025-01293-9

Figure Lengend Snippet: A Groups of male and female C57BL/6J mice received either prime-only or prime-boost immunization with 1 × 10 5 TCID 50 of CCHFV VRP (subcutaneous [SC]). At designated timepoints following vaccination (prime-only or prime-boost: 6, 21, 28, or 42 days [D]; and 2, 3, 4, 5, 6, 9, 12, or 18 months [M]) whole blood was collected from the submandibular vein (vaccinated and unvaccinated controls: n = 10–20 [5–10 male, 5–10 female], each) for assessment of humoral immunity. Lethal challenge studies were conducted with cohorts of vaccinated (prime-only: n = 6–16; prime-boost: n = 8 per group) and unvaccinated control mice ( n = 6–12 per group) at a subset of timepoints used to evaluate humoral responses [(2, 4, 6, 9, and 12M (prime-only or prime-boost) and 18M (prime-only)]. On the day of challenge (0 dpi) mice were transiently immunosuppressed with 2.5 mg of anti-mouse IFNAR1 MAR1-5A3 monoclonal antibody (intraperitoneally [IP]), challenged SC with a target dose of 1 × 10 TCID 50 of CCHFV strain IbAr10200, and followed for up to 14 days post-infection (dpi). B Anti-NP IgG antibody response kinetics were quantified and included anti-nucleoprotein (NP) antibody titer, subclass composition (IgG1 and IgG2c), avidity, and the Fc-mediated effector functions antibody-dependent complement deposition (ADCD) and antibody-dependent cellular phagocytosis (ADCP). Each circle represents an individual animal; points are aligned for clarity, and some overlap may occur. The solid line indicates the mean. Where appropriate, the dashed grey line indicates the assay mean from control naïve sera (unvaccinated), and the shaded grey area represents ±1 standard deviation. See Supplemental Figs. and for statistical analyses.

Article Snippet: At the time of challenge (day 0) mice were transiently immunosuppressed with 2.5 mg of anti-mouse IFNAR1 monoclonal antibody administered intraperitoneally (MAR1-5A3; Leinco Technologies; Cat. No. I-401, Lot No. 1223L560, 0823L230, 0822L285, and 0323L565).

Techniques: Control, Infection, Standard Deviation

Journal: bioRxiv

Article Title: ADAR1p150 RNA binding, independent of A-to-I RNA editing, buffers immunogenicity from the tonic type I IFN induced transcriptome in vivo

doi: 10.1101/2025.10.14.682456

Figure Lengend Snippet: ( A ) Representative FACS plots from an Adar1 +/+ Ifih1 -/- Eif2ak2 -/- ( A1 +/+ ) showing the gating strategy to identify the HSC and progenitor fraction. ( B ) Representative FACS plots from an Adar1 p150-/- Ifih1 -/- Eif2ak2 -/- ( A1 p150-/- ) animal. Experiment repeated 4 independent times with 4 pairs of control and A1 p150-/- males each time. ( C ) Frequency of each population in each genotype; each data point represents as individual animal and mean ± SEM as indicated, n=4 per genotype. ( D ) Representative microscope images for each indicated cell population in each treatment arm. ( E ) The cell number for each population expressed as a percentage of untreated cells (left panel) and viability as determined by trypan blue cell counting expressed as a percentage of untreated cells (right panel); data expressed as mean ± SEM, n=4 per genotype. ( F ) Schematic outline of in vivo experiment to block IFNAR1 using monoclonal antibody administration. ( G ) Body weight of each animal administered αIgG 1 or αIFNAR1 monoclonal antibody in each experiment. Each experiment was performed with an independent batch of αIFNAR1 with n per genotype as indicated. ( H ) Representative FACS plots of peripheral blood T cell populations. ( I ) Quantitation of the peripheral blood CD8 (left panel) and CD4 (right panel) T cells in each genotype and treatment cohort; Data from each individual animal is linked by a line and all animals across both replicates pooled. Statistical significance determined by multiple paired T test with FDR correction for multiple comparisons. ( J ) Representative FACS plots of bone marrow HSPC populations; populations gated on lineage - cKit + Sca1 + CD135 - population. (K) Frequency of each indicated population in each genotype and treatment cohort; each data point represents as individual animal and mean ± SEM as indicated and all animals across both replicates pooled, Statistical significance determined by multiple paired t-tests with FDR correction for multiple comparisons.

Article Snippet: Mice were weighed and treatment commenced with anti-IFNAR1 monoclonal antibody (MAR1-5A3; isotype mouse IgG 1 ; Leinco Technologies Inc) or anti-mouse IgG 1 control (Isotype Control; Leinco Technologies Inc) , .

Techniques: Control, Microscopy, Cell Counting, In Vivo, Blocking Assay, Quantitation Assay

Journal: bioRxiv

Article Title: ADAR1p150 RNA binding, independent of A-to-I RNA editing, buffers immunogenicity from the tonic type I IFN induced transcriptome in vivo

doi: 10.1101/2025.10.14.682456

Figure Lengend Snippet: ( A ) Replicate independent experiment to data in on A1 triple myeloid cells treated with 50U/mL rmIFNβ; data presented as percentage of live cells (compared to untreated time 0; trypan blue staining) following treatment for 48 hours; each individual cell line indicated by a dot; statistics calculated using 2-way ANOVA with Tukey’s multiple comparison correction. ( B ) Comparisons by indicated genotype (using same data as panel A). ( C ) Viable cell number of each indicated genotype treated with 50U/mL rmIFNβ for 48 hours; each individual cell line indicated by a dot; statistics calculated using 2-way ANOVA with Tukey’s multiple comparison correction. ( D ) Cell surface IFNAR1 expression on the myeloid cell lines compared to anti-IgG 1 κ PE isotype control; each profile is from an independent cell line of the indicated genotype and quantitation and analysis of the mean fluorescence index of IFNAR1 expression (based on analysis in FlowJo). ( E ) Transcript counts of Ifnar1 , Ifnar2 , Ifngr1 and Ifngr2 from RNA-seq of untreated and cells treated with 50U/mL rmIFNβ for 48 hours; each individual cell line indicated by a dot. Data expressed as mean ± sem normalized counts per million reads (CPM). ( F-G ) The effect of increasing doses of rmIFNβ on A1 +/+ or A1 p150-/- cells as measured using continuous monitoring with Incucyte across time. Data show increase in cell death with dose and time as measured by Cytotox area over phase normalised area at Time 0. ( H ) Effect of 50U/mL rmIFNβ treatment on the indicated genotypes for up to 7 days; Cell counts and viability from Countess/Trypan blue. Cell lines - A1 +/+ = 213 (derived from mouse #213); A1 -/- = 616; A1 p150-/- = 207. ( I ) Treatment of cells with 25U/mL rmIFNψ. Each individual cell line indicated by a dot; statistics calculated using 2-way ANOVA with Tukey’s multiple comparison correction. ***p<0.001.

Article Snippet: Mice were weighed and treatment commenced with anti-IFNAR1 monoclonal antibody (MAR1-5A3; isotype mouse IgG 1 ; Leinco Technologies Inc) or anti-mouse IgG 1 control (Isotype Control; Leinco Technologies Inc) , .

Techniques: Staining, Comparison, Expressing, Control, Quantitation Assay, Fluorescence, RNA Sequencing, Derivative Assay

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