IFNAR1 Antibody

What is IFNAR1 and what is its role in immune signaling?

IFNAR1 (interferon alpha and beta receptor subunit 1) is a critical component of the type I interferon receptor complex. It functions as a transmembrane receptor that binds type I interferons (including IFN-alpha, beta, and omega), initiating a signaling cascade vital for antiviral responses, immune regulation, and cellular growth control. IFNAR1 has a molecular weight of approximately 63.5 kilodaltons and forms a heterodimeric receptor complex with IFNAR2 to facilitate signal transduction through JAK-STAT pathways . This signaling activates transcription of interferon-stimulated genes (ISGs) that mediate various biological effects, including antiviral, antiproliferative, and immunomodulatory functions.

How does IFNAR1 signaling affect immune checkpoint pathways?

Research demonstrates that IFNα can actually promote immunosuppression through IFNAR1 signaling by transcriptionally activating programmed death ligand 1 (PDL1) expression through phosphorylated STAT1 (Tyr701). Additionally, IFNAR1 signaling can promote programmed cell death protein 1 (PD1) expression in immune cells. This represents a previously underappreciated mechanism of immunosuppression, particularly relevant in head and neck squamous cell carcinomas (HNSCC) . The dual upregulation of both PDL1 on tumor cells and PD1 on immune cells creates a potent immunosuppressive environment that facilitates tumor immune evasion.

What is Anifrolumab and how does it relate to IFNAR1?

Anifrolumab is an FDA-approved monoclonal antibody that specifically targets IFNAR1. It functions by binding to and blocking the type I interferon receptor, thereby inhibiting type I IFN signaling . Originally developed for treating systemic lupus erythematosus (SLE), Anifrolumab represents a significant therapeutic advance in targeting the type I interferon pathway. For research purposes, Anifrolumab and its biosimilars serve as valuable tools for studying IFNAR1 biology and developing experimental models of type I interferon blockade .

What are the primary applications for IFNAR1 antibodies in research?

IFNAR1 antibodies can be employed across multiple experimental platforms including:

• Western blot (WB) for protein expression analysis

• Immunoprecipitation (IP) for protein complex studies

• Flow cytometry (FCM) for cell surface expression analysis

• Immunocytochemistry (ICC) and immunofluorescence (IF) for localization studies

• Immunohistochemistry (IHC) for tissue expression patterns

• Neutralization assays to block IFN-α/β signaling

• ELISA for quantitative detection

The choice of application depends on the specific research question and experimental system being investigated.

How should researchers optimize Western blot protocols for IFNAR1 detection?

For optimal IFNAR1 detection via Western blot, consider the following methodological approach:

• Sample preparation: Use RIPA or NP-40 lysis buffers with protease inhibitors

• Protein loading: 20-50μg total protein per lane depending on expression level

• Gel selection: 8-10% SDS-PAGE gels for optimal resolution of the 63.5 kDa IFNAR1 protein

• Transfer conditions: Semi-dry or wet transfer to PVDF membranes (preferred over nitrocellulose)

• Blocking: 5% non-fat milk or BSA in TBST for 1-2 hours

• Primary antibody: Dilute according to manufacturer recommendations (typically 1:500-1:2000)

• Incubation: Overnight at 4°C with gentle rocking

• Secondary antibody: Use species-appropriate HRP-conjugated secondary antibody

• Detection: Enhanced chemiluminescence with appropriate exposure time

When selecting between the multiple commercially available antibodies, prioritize those with demonstrated specificity in Western blot applications and relevant citations in systems similar to yours .

What controls should be included when using IFNAR1 antibodies for functional studies?

Rigorous experimental design for IFNAR1 antibody studies should include:

• Positive control: Cell lines with confirmed high IFNAR1 expression (e.g., certain immune cell lines)

• Negative control: IFNAR1 knockout cells or cells treated with IFNAR1 siRNA

• Isotype control antibody: Matching isotype (e.g., human IgG1 for Anifrolumab-based studies)

• Specificity validation: Peptide competition assay to confirm antibody specificity

• Functional validation: Confirm blocking activity using a reporter cell line such as HEK-Blue™ IFN-α/β cells to measure inhibition of IFN-α/β signaling

Including these controls ensures the validity of experimental findings and facilitates accurate interpretation of results.

How can IFNAR1 antibodies be used to study immunosuppressive mechanisms in cancer?

Researchers investigating cancer immunology can employ IFNAR1 antibodies to:

• Characterize the paradoxical immunosuppressive effects of type I IFNs in the tumor microenvironment

• Investigate the relationship between IFNAR1 signaling and immune checkpoint expression

• Develop strategies to counteract IFNα-induced PDL1 and PD1 expression

Methodologically, this involves treating cancer cells with type I IFNs (IFNα or IFNβ) followed by analysis of PDL1 expression, then using IFNAR1-blocking antibodies to demonstrate the specificity of this effect. Studies in HNSCC have shown that IFNα transcriptionally activates PDL1 expression through p-Stat1 (Tyr701) and promotes PD1 expression in immune cells through IFNAR1 . This approach has revealed that inhibition of IFNα signaling can enhance the cytotoxic activity of natural killer cells, suggesting a potential therapeutic strategy.

What are the methodological considerations when using IFNAR1 antibodies in patient-derived xenograft models?

When employing IFNAR1 antibodies in patient-derived xenograft (PDX) models, researchers should consider:

• Species specificity: Ensure the anti-IFNAR1 antibody recognizes human IFNAR1 if studying human tumors in immunocompromised mice

• Dosing regimen: Establish appropriate dosing based on antibody half-life and previous literature

• Administration route: Intraperitoneal or intravenous injection depending on model requirements

• Sampling timeline: Collect tissues at multiple timepoints to assess dynamic changes

• Combined analyses: Integrate immunohistochemistry, flow cytometry, and functional assays

Research has demonstrated the utility of this approach, with upregulation of PDL1 and PD1 in response to IFNα treatment confirmed in both conventional xenograft tumor models and patient-derived xenograft models . This provides a system to test combination therapies involving IFNAR1 blockade and other immunomodulatory approaches.

How can researchers analyze the impact of IFNAR1 blockade on interferon-stimulated gene expression?

To comprehensively assess the effects of IFNAR1 blockade on downstream gene expression:

• Treat cells with type I IFNs with or without IFNAR1-blocking antibodies

• Extract RNA at multiple timepoints (e.g., 2, 6, 12, 24 hours)

• Perform qRT-PCR for key interferon-stimulated genes (ISGs) such as MX1, OAS1, and STAT1

• Alternatively, conduct RNA-sequencing for genome-wide ISG expression analysis

• Validate protein-level changes via Western blot or proteomics approaches

• Correlate gene expression changes with functional outcomes in relevant assays

This approach allows researchers to establish the temporal dynamics of IFNAR1-mediated signaling and identify key regulatory nodes that might represent therapeutic targets. Evidence of endogenous IFNα activation in tumor microenvironments has been associated with overexpression of IFNAR1, MX1, and STAT1, correlating with immunosuppression status in HNSCC patients .

What are common pitfalls when using IFNAR1 antibodies and how can they be addressed?

Several technical challenges may arise when working with IFNAR1 antibodies:

When selecting an anti-IFNAR1 antibody, prioritize products with demonstrated specificity and relevant citations in applications similar to your experimental design .

How should researchers validate the specificity of IFNAR1 antibodies?

Comprehensive validation strategy for IFNAR1 antibodies should include:

• Positive and negative cell lines: Compare detection in cells known to express IFNAR1 versus those that do not

• Knockdown/knockout validation: Test antibody in IFNAR1 siRNA-treated or CRISPR-Cas9 knockout cells

• Peptide competition assay: Pre-incubate antibody with immunizing peptide to block specific binding

• Multiple application testing: Confirm consistent results across different experimental platforms

• Orthogonal methods: Correlate protein detection with mRNA expression via qRT-PCR

• Cross-reactivity assessment: Test on closely related proteins, particularly IFNAR2

What are the key considerations for storage and handling of IFNAR1 antibodies?

To maintain antibody integrity and performance:

• Storage temperature: Follow manufacturer recommendations (typically -20°C for long-term)

• Aliquoting: Divide stock solution into single-use aliquots to avoid freeze-thaw cycles

• Reconstitution: For lyophilized antibodies, use sterile water as specified

• Working dilution: Prepare fresh working dilutions for each experiment

• Stabilizers: Consider adding BSA (0.1-1%) to diluted antibodies for stability

• Contamination prevention: Use sterile technique when handling antibody solutions

• Quality control: Test performance periodically against a reference sample

Many IFNAR1 antibodies are provided in lyophilized form and require reconstitution before use . Optimal reconstitution and storage practices significantly impact experimental reproducibility.

How do IFNAR1 antibodies contribute to research on autoimmune diseases?

IFNAR1 antibodies have become instrumental in studying autoimmune pathologies:

• Mechanistic studies: IFNAR1 blocking antibodies help delineate the role of type I IFN signaling in disease pathogenesis

• Biomarker identification: Anti-IFNAR1 antibodies facilitate detection of receptor expression levels as potential disease biomarkers

• Therapeutic development: Research using anti-IFNAR1 antibodies has led to clinical applications, as evidenced by the FDA approval of Anifrolumab for systemic lupus erythematosus (SLE)

• Patient stratification: Studying IFNAR1 expression and signaling helps identify patient subgroups likely to respond to IFN-targeting therapies

• Preclinical models: IFNAR1 antibodies enable creation of relevant disease models through selective pathway inhibition

The translation of basic research findings to clinical applications demonstrates the significant impact of fundamental studies using IFNAR1 antibodies on patient care.

What methodological approaches are recommended for studying IFNAR1 in cancer immunotherapy research?

For cancer immunotherapy investigations involving IFNAR1:

• Expression analysis: Quantify IFNAR1 levels in tumor vs. normal tissues using IHC, flow cytometry, and Western blot

• Functional assays: Assess the impact of IFNAR1 blockade on immune cell recruitment and activation in tumor models

• Combination studies: Test IFNAR1-blocking antibodies in combination with immune checkpoint inhibitors

• Biomarker correlation: Relate IFNAR1 expression to response to immunotherapy

• Signaling pathway analysis: Evaluate downstream effects on STAT1 phosphorylation and target gene expression

Research has demonstrated that IFNα-induced PDL1 and PD1 expression represents a previously unrecognized mechanism of immunosuppression in HNSCC, suggesting that blocking IFNα signaling may enhance the efficacy of immune checkpoint blockade therapies . This approach could potentially overcome resistance mechanisms in cancer immunotherapy.

How can researchers best study the relationship between IFNAR1 and PDL1/PD1 expression?

To investigate the regulatory relationship between IFNAR1 signaling and immune checkpoint molecules:

• Dose-response studies: Treat cells with increasing concentrations of type I IFNs and measure PDL1/PD1 expression

• Temporal analysis: Determine the kinetics of PDL1/PD1 upregulation following IFN stimulation

• Signaling inhibition: Use IFNAR1 antibodies alongside JAK/STAT inhibitors to delineate the precise signaling cascade

• Chromatin immunoprecipitation (ChIP): Assess p-STAT1 binding to PDL1 promoter regions

• Reporter assays: Construct PDL1 promoter-reporter systems to measure transcriptional activation

• Co-culture systems: Evaluate the functional impact on T cell or NK cell activity

This multimodal approach has revealed that IFNα transcriptionally activates PDL1 expression through p-Stat1 (Tyr701) and promotes PD1 expression in immune cells through IFNAR1 signaling . These findings suggest novel therapeutic strategies combining IFNAR1 blockade with immune checkpoint inhibitors.

What emerging technologies can enhance IFNAR1 antibody-based research?

Several cutting-edge approaches are advancing IFNAR1 research:

• Single-cell analysis: Combining IFNAR1 antibodies with single-cell RNA-seq to understand cellular heterogeneity in response to type I IFNs

• Spatial transcriptomics: Correlating IFNAR1 protein expression with spatial gene expression patterns in tissues

• CRISPR screens: Identifying modulators of IFNAR1 signaling through genome-wide functional screens

• Antibody engineering: Developing bispecific antibodies targeting IFNAR1 and other immune regulators

• Real-time imaging: Using fluorescently-labeled IFNAR1 antibodies for intravital microscopy to track receptor dynamics

• Protein interaction mapping: Applying proximity labeling techniques to identify novel IFNAR1 interaction partners

These technologies promise to reveal new insights into IFNAR1 biology and potential therapeutic applications beyond current understanding.

How might combinatorial approaches using IFNAR1 antibodies advance cancer immunotherapy?

Innovative combination strategies include:

• IFNAR1 blockade + PD1/PDL1 inhibitors: Targeting both IFN-induced immunosuppression and established checkpoint pathways

• IFNAR1 inhibition + radiation therapy: Modulating the IFN response to enhance radiation sensitivity

• Sequential therapy: Temporary IFNAR1 blockade followed by checkpoint inhibition to reshape the tumor microenvironment

• Cell-specific targeting: Developing approaches to selectively inhibit IFNAR1 on specific cell populations

• Biomarker-guided combinations: Using IFNAR1 expression or IFN signature as predictive biomarkers for combination selection

Research in HNSCC models has demonstrated that inhibition of IFNα signaling enhances the cytotoxic activity of natural killer cells, suggesting that blocking IFNα signaling may enhance the efficacy of immune checkpoint blockade . This provides a foundation for rational design of combination immunotherapies targeting the IFN pathway.

What are the key considerations for developing next-generation therapeutic IFNAR1 antibodies?

Advancing IFNAR1-targeted therapeutics requires addressing:

• Receptor subtype selectivity: Engineering antibodies with specificity for distinct functional epitopes

• Tissue-specific targeting: Developing delivery systems that target specific anatomical sites

• Reduced immunogenicity: Optimizing antibody humanization to minimize anti-drug antibody responses

• Pharmacokinetic enhancement: Modifying Fc regions to extend half-life and improve tissue penetration

• Effector function engineering: Tailoring Fc-mediated functions for specific therapeutic contexts

• Combination potential: Designing antibodies compatible with standard-of-care treatments

Anifrolumab's success in SLE treatment provides a foundation for developing next-generation anti-IFNAR1 therapeutics with enhanced efficacy and safety profiles . These developments will build on fundamental research utilizing IFNAR1 antibodies in experimental settings.