PILRA Antibody

What is PILRA and why is it significant in immunological research?

PILRA (paired immunoglobulin-like type 2 receptor alpha) is an immune inhibitory receptor possessing an immunoreceptor tyrosine-based inhibitory motif (ITIM) in its cytoplasmic domain that enables it to deliver inhibitory signals. PILRA belongs to the paired receptor family that consists of highly related activating and inhibitory receptors widely involved in immune system regulation . Its significance in immunological research stems from its role in modulating immune responses by interacting with ligands including CD99, PILR-associating neural protein (PANP), and HSV-1 glycoprotein B . Recent research has revealed PILRA's involvement in neuroinflammation processes relevant to Alzheimer's disease and its potential roles in cancer immunity, making it an increasingly important target for therapeutic development .

What are the structural and biochemical characteristics of PILRA protein?

PILRA is characterized by:

• Calculated molecular weight: 34 kDa

• Observed molecular weight: 44-50 kDa (due to glycosylation)

• Length: 303 amino acids

• Gene ID (NCBI): 29992

• UniProt ID: Q9UKJ1

The protein features extracellular ligand-binding domains similar to its paired receptor PILRβ, but contains divergent intracellular signaling domains. The presence of the ITIM motif in its cytoplasmic domain is critical for its inhibitory function in immune regulation . The discrepancy between calculated and observed molecular weights is attributed to post-translational modifications, particularly glycosylation, which affects the protein's migration pattern in gel electrophoresis .

How do PILRA antibodies differ in their applications for immunological research?

PILRA antibodies are designed for different research applications with varying specificity and sensitivity profiles. Commercial antibodies like 11818-1-AP have been validated for Western Blot (WB) and ELISA applications with recommended dilutions of 1:500-1:2000 for WB . Monoclonal antibodies such as MAB64841 have demonstrated efficacy in multiple applications including:

Each antibody may require optimization depending on the specific experimental conditions and sample types . Therapeutic antibodies against PILRA are being developed to mimic the effects of loss-of-function variants that appear protective against Alzheimer's disease .

How does PILRA function relate to Alzheimer's disease pathology?

PILRA has emerged as a significant factor in Alzheimer's disease (AD) pathology through several mechanisms:

• Loss-of-function variants in the PILRA gene, particularly the common G78R variant (SNP rs1859788), protect against AD development .

• In microglia, PILRA knockout induces metabolic changes including:

  • Enhanced mitochondrial respiration and ATP production

  • Reduced inflammatory cytokine production

  • Improved protein degradation through lysosomal activity

  • Altered lipid metabolism favoring antioxidant lipids

These changes suggest that PILRA normally constrains microglial function, and its inhibition may represent a therapeutic approach for AD. Researchers at Denali Therapeutics have developed an anti-PILRA antibody that blocks ligand binding and replicates the beneficial effects observed in PILRA knockout models, including boosted mitochondrial respiration, tempered cytokine production, and enhanced cell migration .

What experimental models are most effective for studying PILRA's role in neuroinflammation?

Current research indicates several effective experimental models for investigating PILRA in neuroinflammation:

• Human iPSC-derived microglia (iMG): These provide a physiologically relevant human cellular model for studying PILRA function. Knockout experiments in these cells have revealed significant changes in transcriptome, metabolome, and lipidome profiles related to PILRA function .

• Chimeric mouse models: Xenotransplantation models where murine microglia are replaced with human PILRA knockout microglia allow for in vivo assessment of PILRA's functions. These models enable researchers to study effects on amyloid pathology and neuroinflammation in a complex central nervous system environment .

• Antibody-based intervention models: Treatment with anti-PILRA antibodies in cell culture or animal models can mimic genetic loss-of-function and provide insights into potential therapeutic approaches .

For optimal results, researchers should consider combining these models to gain comprehensive insights into both cellular mechanisms and systemic effects of PILRA modulation in neuroinflammatory contexts .

How do APOE and PILRA genotypes interact in Alzheimer's disease risk profiles?

Recent studies have identified a complex interaction between APOE genotype and PILRA variants in modulating Alzheimer's disease risk:

• PILRA SNP rs1859788 has been confirmed to reduce AD risk, with a particularly significant association observed in men who are homozygous for the APOE ε4/ε4 isoform .

• Both PILRA and APOE are involved in key AD pathological processes:

  • Amyloid-beta metabolism

  • Neuroinflammation regulation

  • Glial cell activation

• Mechanistically, PILRA may modulate the inflammatory response to amyloid-beta accumulation, potentially affecting the severity of inflammation and subsequent neuronal damage in APOE ε4 carriers .

This interaction suggests potential for personalized medicine approaches that consider both PILRA and APOE status when developing therapeutic strategies. Understanding these genetic interactions provides insights into AD heterogeneity and may help identify novel therapeutic targets specific to certain genetic profiles .

What is the significance of PILRA expression in cancer immunosurveillance?

PILRA has emerged as a potentially important factor in cancer immunity with significant implications for immunosurveillance mechanisms:

• Pan-cancer analysis has revealed that PILRA is significantly dysregulated and frequently mutated across multiple cancer types .

• Both PILRA expression levels and mutation status have been shown to significantly impact patient prognosis in several cancers, suggesting its role in modulating anti-tumor immune responses .

• PILRA expression positively correlates with:

  • ESTIMATE scores (reflecting tumor immune infiltration)

  • Abundance of tumor-infiltrating immune cells

  • Predictive biomarkers of cancer immunotherapy

  • Prognostic outcomes in patients receiving immunotherapy

These findings suggest that PILRA may function as an immune checkpoint molecule in the tumor microenvironment, potentially regulating the efficacy of anti-tumor immune responses. The correlation between PILRA expression and immunotherapy outcomes makes it a promising biomarker and potential therapeutic target for cancer immunotherapy optimization .

How can researchers effectively analyze PILRA's role in tumor microenvironment and immune cell infiltration?

To effectively analyze PILRA's role in the tumor microenvironment and immune cell infiltration, researchers should employ a multi-faceted approach:

• Transcriptomic analysis: Utilize RNA-sequencing data from resources like The Cancer Genome Atlas (TCGA) to analyze PILRA expression across cancer types and correlate with immune signatures .

• Immune cell deconvolution: Apply computational methods such as CIBERSORT, xCell, or MCP-counter to estimate immune cell populations from bulk RNA-seq data and correlate with PILRA expression .

• Spatial transcriptomics and immunohistochemistry: Combine PILRA antibody staining with immune cell markers to visualize spatial relationships between PILRA-expressing cells and tumor-infiltrating immune cells. This approach has been validated in tissues such as human tonsil where PILRA expression localizes to dendritic cells .

• Functional assays: Implement PILRA knockout or antibody blockade in co-culture systems with cancer cells and immune cells to assess functional consequences on immune activation, cytokine production, and cancer cell killing .

• Correlation with immunotherapy biomarkers: Analyze relationships between PILRA expression and established immunotherapy response biomarkers such as PD-L1 expression, tumor mutational burden, and microsatellite instability status .

By integrating these approaches, researchers can comprehensively characterize how PILRA influences the tumor immune microenvironment and potentially identify patient populations who might benefit from PILRA-targeted therapies .

What are the optimal conditions for using PILRA antibodies in Western blot applications?

For optimal Western blot results with PILRA antibodies, researchers should consider the following protocol recommendations:

• Sample preparation:

  • Successfully detected in human PBMCs, spleen tissue, lung tissue, and cell lines including CEM and U937

  • Use RIPA buffer with protease inhibitors for efficient protein extraction

  • Load 20-30 μg of total protein per lane

• Electrophoresis conditions:

  • Use reducing conditions for best results

  • Expect PILRA to migrate between 35-50 kDa (significantly higher than the calculated 34 kDa due to glycosylation)

• Transfer and blocking:

  • PVDF membrane is recommended over nitrocellulose

  • Block with 5% non-fat milk in TBST for 1 hour at room temperature

• Antibody incubation:

  • Primary antibody dilution ranges from 1:500-1:2000 for polyclonal antibodies

  • For monoclonal antibodies, 2 μg/mL concentration has been validated

  • Incubate overnight at 4°C for best signal-to-noise ratio

• Detection system:

  • HRP-conjugated secondary antibodies work well with PILRA detection

  • Use Immunoblot Buffer Group 1 for optimal results with MAB64841

It's important to note that glycosylation patterns may vary between cell types and sample conditions, potentially affecting the observed molecular weight. Therefore, validation with positive controls is recommended when working with new sample types .

How can researchers troubleshoot inconsistent PILRA antibody staining in immunohistochemistry?

When troubleshooting inconsistent PILRA antibody staining in immunohistochemistry (IHC), researchers should systematically address several key factors:

• Tissue fixation and processing:

  • Optimal fixation: Use 10% neutral buffered formalin for 24-48 hours

  • Over-fixation can mask epitopes, requiring more aggressive antigen retrieval

  • Paraffin embedding provides better morphology than frozen sections for PILRA detection

• Antigen retrieval optimization:

  • Heat-induced epitope retrieval (HIER) using citrate buffer (pH 6.0) is generally effective

  • For difficult samples, try alternative buffers (Tris-EDTA, pH 9.0)

  • Optimize retrieval time (10-30 minutes) based on tissue type and fixation conditions

• Antibody selection and validation:

  • Monoclonal antibodies like MAB64841 have shown specific staining in human tonsil tissue, particularly in dendritic cells

  • Always include positive controls (e.g., human tonsil tissue) alongside experimental samples

  • Negative controls should include isotype antibodies and PILRA-negative tissues

• Detection system considerations:

  • Polymer-based detection systems (e.g., VisUCyte™ HRP Polymer) provide high sensitivity with low background

  • DAB (3,3'-diaminobenzidine) chromogen produces stable brown staining ideal for PILRA visualization

  • Consider tissue autofluorescence when selecting between chromogenic and fluorescent detection methods

• Troubleshooting specific issues:

  • Weak signal: Increase antibody concentration, extend incubation time, or enhance detection system

  • High background: Increase blocking time/concentration, reduce antibody concentration, include additional washing steps

  • Non-specific staining: Validate antibody specificity using knockout controls or peptide competition assays

By systematically addressing these factors, researchers can achieve consistent and specific PILRA staining in IHC applications .

What controls are essential when validating PILRA knockout or knockdown experiments?

When validating PILRA knockout or knockdown experiments, implementing comprehensive controls is crucial for experimental rigor and reproducibility:

• Genetic validation controls:

  • Sequence verification of the targeted PILRA locus in engineered cells

  • Multiple guide RNAs or siRNAs targeting different regions of PILRA to rule out off-target effects

  • Analysis of potential compensatory changes in related genes (especially PILRB expression)

• Protein expression validation controls:

  • Western blot analysis using validated PILRA antibodies to confirm protein depletion

  • Flow cytometry to assess cell surface PILRA expression in individual cells

  • Immunofluorescence to visualize subcellular localization changes

• Functional validation controls:

  • Rescue experiments with wild-type PILRA to confirm phenotype specificity

  • Comparison with pharmacological inhibition (e.g., anti-PILRA antibody treatment)

  • Analysis of established PILRA downstream signaling pathways

• Experimental design controls:

  • Wild-type isogenic controls processed in parallel

  • Empty vector or non-targeting guide RNA/siRNA controls

  • Multiple independent knockout/knockdown clones or populations to account for clonal variation

• Phenotypic assay controls:

  • For microglia studies: assess metabolic parameters, inflammatory cytokine production, and migration capacity

  • For cancer studies: measure immune cell interactions and cytokine production

  • Positive controls known to affect the same pathways (e.g., other inhibitory receptors)

Researchers studying PILRA in the context of Alzheimer's disease should particularly focus on validating microglial activation states, mitochondrial function, and inflammatory responses, as these parameters have been shown to be significantly altered in PILRA knockout models .

How do glycosylation patterns of PILRA affect antibody binding and functional outcomes?

The glycosylation status of PILRA represents a critical factor affecting both antibody recognition and functional activity:

PILRA has a calculated molecular weight of 34 kDa but typically migrates at 44-50 kDa in gel electrophoresis due to glycosylation . This substantial difference suggests extensive post-translational modification that may have functional significance. Research considerations regarding PILRA glycosylation include:

• Impact on antibody recognition:

  • Glycan structures may mask or alter epitopes recognized by different antibodies

  • Deglycosylation experiments with enzymes like PNGase F can help distinguish antibodies that recognize glycan-dependent versus protein-backbone epitopes

  • Multiple antibodies targeting different epitopes should be used when analyzing PILRA in different cellular contexts

• Functional consequences:

  • Glycosylation patterns may alter PILRA's binding affinity for its ligands including CD99, PANP, and HSV-1 gB

  • Cell-type specific glycosylation could create functional heterogeneity across different immune cell populations

  • Disease states may alter glycosylation patterns, potentially affecting PILRA inhibitory function

• Experimental considerations:

  • Researchers should be aware that reducing conditions in Western blotting may affect glycoprotein migration

  • Expression systems used for recombinant PILRA production may not recapitulate native glycosylation patterns

  • Mass spectrometry approaches can be used to characterize site-specific glycosylation profiles

Understanding the relationship between PILRA glycosylation, antibody binding, and functional outcomes will be crucial for developing effective therapeutic antibodies and interpreting research findings across different experimental systems .

What are the mechanisms behind PILRA's differential effects in neurodegenerative versus oncogenic contexts?

PILRA appears to play context-dependent roles in neurodegenerative diseases and cancer, suggesting complex underlying mechanisms:

• In neurodegenerative contexts:

  • Loss-of-function PILRA variants are protective against Alzheimer's disease

  • PILRA inhibition in microglia leads to:

    • Enhanced mitochondrial function and ATP production

    • Reduced inflammatory cytokine production

    • Improved protein degradation capacity

    • Metabolic shifts favoring neuroprotective functions

• In oncogenic contexts:

  • PILRA expression correlates with immune infiltration in tumors

  • PILRA may function as an immune checkpoint molecule

  • PILRA expression associates with immunotherapy response

  • Both expression levels and mutation status affect cancer prognosis

• Potential explanatory mechanisms:

  • Tissue-specific ligand expression patterns may drive different outcomes

  • Microenvironmental factors (inflammatory vs. immunosuppressive) may alter PILRA signaling

  • Cell-specific PILRA expression levels may reach different thresholds for inhibitory function

  • Interaction with distinct downstream signaling networks in different cell types

• Convergent pathways:

  • Both contexts involve PILRA's role in regulating inflammatory responses

  • Metabolic modulation appears central to PILRA function in both settings

  • Immune cell activation status is affected by PILRA in both disease contexts

Future research should focus on comparative studies examining PILRA function across these contexts, potentially revealing therapeutic strategies that could be tailored to either promote or inhibit PILRA activity depending on the disease context .

How might targeting PILRA with therapeutic antibodies affect age-related conditions beyond Alzheimer's disease?

Given PILRA's emerging role in immune regulation and neuroinflammation, therapeutic antibodies targeting this receptor could potentially impact various age-related conditions:

• Other neurodegenerative disorders:

  • Parkinson's disease, where microglial activation contributes to pathology

  • Frontotemporal dementia, which shares inflammatory components with AD

  • Amyotrophic lateral sclerosis, where altered immune function affects disease progression

• Age-related inflammatory conditions:

  • Research has revealed connections between PILRA and megaloblastic anemia

  • PILRA modulation could potentially affect other age-related inflammatory conditions such as:

    • Chronic low-grade inflammation ("inflammaging")

    • Autoimmune disorders more prevalent in elderly populations

    • Age-related macular degeneration, which involves microglial dysfunction

• Therapeutic considerations:

  • Anti-PILRA antibodies are being developed with blood-brain barrier penetration capabilities through transferrin receptor targeting ("antibody transport vehicle")

  • Combination approaches with anti-amyloid therapies are being explored for AD

  • Similar combination strategies could be relevant for other age-related pathologies

• Potential mechanisms of action in age-related conditions:

  • Enhanced mitochondrial function in immune cells, potentially counteracting age-related bioenergetic decline

  • Reduced inflammatory cytokine production, addressing a hallmark of aging

  • Improved protein degradation capacity, potentially affecting age-related proteostasis defects

As research progresses, therapeutic strategies targeting PILRA will likely need to balance beneficial immune modulation against potential adverse effects, particularly given PILRA's role in regulating responses to certain pathogens .

What are the key considerations when selecting between monoclonal and polyclonal PILRA antibodies for specific applications?

When selecting between monoclonal and polyclonal PILRA antibodies, researchers should consider several application-specific factors:

For PILRA specifically:

• Monoclonal antibodies like MAB64841 have been validated for Western blot, flow cytometry, and IHC applications, demonstrating good specificity across multiple sample types

• Polyclonal antibodies like 11818-1-AP have shown utility in Western blot applications with recommended dilutions of 1:500-1:2000

• Consider the target region - antibodies recognizing the extracellular domain (Gln20-Thr196) are suitable for detecting cell surface PILRA in live cells

The experimental question should guide selection - for functional blocking studies, monoclonals with characterized epitope specificity are preferred, while for initial protein detection in new sample types, polyclonals may offer greater detection probability .

How can researchers effectively design experiments to investigate PILRA-ligand interactions?

Designing robust experiments to investigate PILRA-ligand interactions requires careful consideration of multiple factors:

• Protein expression and purification:

  • Express the extracellular domain of PILRA (Gln20-Thr196) as a recombinant protein

  • Consider using mammalian expression systems to maintain native glycosylation patterns

  • Include appropriate tags (His, Fc) for purification while ensuring they don't interfere with binding

  • Prepare both wild-type PILRA and the G78R variant to compare binding properties

• Binding assay selection:

  • Surface Plasmon Resonance (SPR) for real-time kinetic analysis of PILRA-ligand interactions

  • Enzyme-Linked Immunosorbent Assay (ELISA) for high-throughput screening

  • Bio-Layer Interferometry (BLI) for label-free interaction analysis

  • Flow cytometry using fluorescently-labeled PILRA to detect binding to cell-surface ligands

• Ligand considerations:

  • Include established PILRA ligands as positive controls: CD99, PANP, HSV-1 gB

  • Test binding in both directions (immobilized PILRA vs. immobilized ligand)

  • Use appropriate negative controls (non-binding proteins of similar size/structure)

  • Consider testing tissue-specific ligands relevant to your research context

• Functional validation:

  • Cell-based reporter assays measuring ITIM-dependent signaling

  • Competition assays with anti-PILRA antibodies to confirm binding specificity

  • Mutagenesis studies to identify critical binding residues

  • Co-immunoprecipitation experiments to confirm complex formation in cellular contexts

• Physiological relevance:

  • Test binding under varying pH and ionic conditions relevant to different cellular compartments

  • Investigate how inflammation or cellular activation affects ligand expression and binding

  • Consider the effects of glycosylation on binding by using enzymatic deglycosylation

These approaches can be particularly valuable when evaluating therapeutic antibodies designed to block PILRA-ligand interactions, as is being explored for Alzheimer's disease treatment .

What technical challenges exist in studying PILRA expression in primary human microglia and how can they be overcome?

Studying PILRA expression in primary human microglia presents several technical challenges that require specialized approaches:

• Tissue access and sample quality:

  • Human brain tissue availability is limited and often represents end-stage disease

  • Postmortem delay can affect protein quality and RNA integrity

  • Solution: Establish collaborations with brain banks that minimize postmortem delay; consider rapid autopsy programs; implement strict quality control for RNA integrity number (RIN) values

• Microglial isolation challenges:

  • Microglia represent a small percentage of total brain cells

  • Traditional isolation methods may activate microglia, altering their phenotype

  • Solution: Use gentle mechanical dissociation followed by immunomagnetic separation with microglial markers (CD11b, CD45); perform isolations at 4°C with RNA stabilization reagents; consider single-cell approaches to avoid activation artifacts

• Expression level detection:

  • PILRA expression may be heterogeneous across microglial subpopulations

  • Protein detection requires sensitive methods due to potentially low expression levels

  • Solution: Implement droplet digital PCR for accurate quantification; use high-sensitivity Western blotting protocols; employ flow cytometry with signal amplification for protein detection

• Alternative approaches:

  • iPSC-derived microglia models: As demonstrated in recent PILRA research, human iPSC-derived microglia provide a renewable source of cells for mechanistic studies

  • Xenotransplantation models: Chimeric mice with human microglia allow for in vivo assessment of PILRA function in a complex environment

  • Single-cell RNA sequencing: Enables characterization of PILRA expression across microglial subtypes and activation states

• Validation strategies:

  • Compare findings across multiple methodologies (RNA, protein)

  • Include appropriate controls (isotype antibodies, non-microglial CNS cells)

  • Validate antibodies for specificity using PILRA knockout controls

  • Correlate with established microglial markers (IBA1, TMEM119, P2RY12)

By addressing these challenges with appropriate methodological approaches, researchers can generate more reliable data on PILRA expression and function in human microglia, particularly in the context of neurodegenerative diseases where PILRA variants appear to play significant roles .

How does PILRA function integrate with other risk genes in Alzheimer's disease pathogenesis?

PILRA functions within a complex genetic landscape influencing Alzheimer's disease pathogenesis:

• APOE interactions:

  • PILRA SNP rs1859788 shows particularly significant protective effects in men who are homozygous for the APOE ε4/ε4 isoform

  • Both genes influence key pathological processes including amyloid-beta metabolism and neuroinflammation

  • PILRA may modulate the inflammatory response to amyloid-beta accumulation in an APOE-dependent manner

• Microglial gene networks:

  • PILRA functions within the context of other microglial risk genes for AD including:

    • TREM2: Another innate immune receptor affecting microglial function

    • CD33: Like PILRA, an inhibitory receptor on myeloid cells

    • PLCG2: Involved in immune cell signaling

  • These genes collectively regulate microglial activation states, phagocytosis, and inflammatory responses

• Pathway integration:

  • PILRA modulation affects mitochondrial function and metabolism in microglia

  • This metabolic regulation may intersect with pathways influenced by other risk genes

  • Lipid metabolism changes induced by PILRA knockout may interact with APOE-mediated lipid processing

• Therapeutic implications:

  • Combined targeting of multiple risk pathways may provide synergistic benefits

  • Anti-PILRA antibodies are being considered in combination with anti-amyloid therapies

  • Genetic profiling for PILRA variants alongside other risk genes may inform personalized treatment approaches

Understanding these integrative aspects of PILRA function can inform more comprehensive models of AD pathogenesis and potentially lead to more effective combination therapeutic strategies targeting multiple disease pathways simultaneously .

What role might PILRA play in the relationship between megaloblastic anemia and Alzheimer's disease risk?

Recent research has uncovered intriguing connections between PILRA, megaloblastic anemia, and Alzheimer's disease:

• Epidemiological connections:

  • A phenome-wide association study (PheWAS) revealed that PILRA SNP rs1859788 is associated with megaloblastic anemia

  • Individuals with anemia have been reported to have a 41% increased risk of developing AD

  • People with AD are often anemic, though not specifically iron deficient

• Potential mechanistic links:

  • Megaloblastic anemia typically results from vitamin B12 or folate deficiency

  • These deficiencies can lead to elevated homocysteine levels, which have been associated with cognitive decline and dementia

  • PILRA's involvement in immune regulation might affect hematopoietic processes

• Diagnostic implications:

  • Pernicious anemia can present with confusion, memory loss, cognitive decline, and be mistaken for AD

  • PILRA genetic screening might help identify individuals with anemia who are at higher risk for developing AD

  • Monitoring for anemia in AD patients might be warranted based on these connections

• Research directions:

  • Investigate whether PILRA expression or function differs in hematopoietic cells of individuals with megaloblastic anemia

  • Determine if vitamin B12/folate supplementation affects PILRA expression or function

  • Explore whether treating anemia in at-risk individuals influences AD progression

This emerging connection between PILRA, megaloblastic anemia, and AD risk highlights the complex interplay between immune function, hematological parameters, and neurodegeneration, suggesting new avenues for both diagnostic approaches and potential therapeutic interventions .

How might single-cell technologies advance our understanding of cell-specific PILRA functions?

Single-cell technologies offer powerful approaches to unravel the complexity of PILRA function across different cell types and disease states:

• Single-cell RNA sequencing (scRNA-seq) applications:

  • Map PILRA expression across diverse immune cell populations with unprecedented resolution

  • Identify previously unrecognized cell subpopulations with distinct PILRA expression patterns

  • Track dynamic changes in PILRA expression during disease progression

  • Correlate PILRA expression with global transcriptional programs to identify functional networks

• Spatial transcriptomics advantages:

  • Preserve tissue architecture while obtaining transcriptomic data

  • Visualize spatial relationships between PILRA-expressing cells and their microenvironment

  • Particularly valuable for understanding PILRA's role in tissue contexts like brain (AD) or tumors (cancer)

  • Can be combined with multiplex immunofluorescence to simultaneously detect protein and RNA

• Mass cytometry (CyTOF) contributions:

  • Simultaneously measure PILRA protein expression alongside dozens of other surface and intracellular proteins

  • Create high-dimensional profiles of PILRA-expressing cells

  • Particularly useful for analyzing phosphorylation events downstream of PILRA activation

  • Can reveal differential signaling patterns in health versus disease states

• Single-cell ATAC-seq insights:

  • Map chromatin accessibility changes associated with PILRA expression

  • Identify potential regulatory elements controlling cell-specific PILRA expression

  • Discover transcription factors involved in PILRA regulation

• Integrated multi-omics approaches:

  • Combine multiple single-cell technologies on the same samples for comprehensive characterization

  • Link PILRA genetic variants to expression and functional outcomes at single-cell resolution

  • CITE-seq approaches can simultaneously measure PILRA at both RNA and protein levels

These technologies are particularly valuable for understanding PILRA's role in heterogeneous contexts like the brain, where diverse microglial states may express and utilize PILRA differently, or in tumors, where PILRA may affect distinct immune cell populations within the tumor microenvironment .

What are the most promising therapeutic applications of PILRA modulation based on current evidence?

Based on current evidence, several promising therapeutic applications of PILRA modulation have emerged:

• Alzheimer's disease treatment:

  • Anti-PILRA antibodies that block ligand binding have shown potential in preliminary studies

  • Coupling anti-PILRA antibodies with blood-brain barrier penetration technology (transferrin receptor-targeting "antibody transport vehicle") is being developed by Denali Therapeutics

  • Combination approaches with anti-amyloid therapies may provide synergistic benefits by addressing both pathological protein accumulation and neuroinflammation

• Cancer immunotherapy enhancement:

  • PILRA expression correlates with immunotherapy response markers

  • Targeting PILRA could potentially enhance anti-tumor immune responses

  • PILRA modulation may be particularly beneficial in cancers with high immune infiltration

• Inflammatory condition management:

  • Given PILRA's role in regulating immune responses, targeting this receptor might be beneficial in conditions characterized by dysregulated inflammation

  • The connection with megaloblastic anemia suggests potential applications in hematological conditions

• Precision medicine approaches:

  • PILRA genotyping could identify individuals most likely to benefit from PILRA-targeting therapies

  • Combined genetic profiling of PILRA and interacting genes like APOE could inform personalized treatment strategies

The most advanced therapeutic development appears to be in Alzheimer's disease, where the protective effects of PILRA loss-of-function have been well-characterized and antibody approaches are being actively developed .

What technical innovations are needed to advance PILRA research in the next five years?

To significantly advance PILRA research over the next five years, several technical innovations and methodological improvements will be crucial:

• Improved blood-brain barrier (BBB) delivery systems:

  • Enhanced antibody transport vehicles beyond the transferrin receptor approach

  • Nanoparticle-based delivery systems for PILRA-targeting therapeutics

  • Non-invasive techniques to temporarily open the BBB for drug delivery

• Advanced in vitro models:

  • Microfluidic "organ-on-chip" systems incorporating multiple cell types to study PILRA in complex tissue environments

  • 3D organoid models with integrated immune components to better recapitulate PILRA function in tissue contexts

  • Patient-derived models that incorporate individual genetic backgrounds including PILRA variants

• Refined animal models:

  • Conditional and inducible PILRA knockout models to study temporal aspects of PILRA function

  • Humanized models expressing human PILRA variants to better translate findings to human disease

  • Multi-gene models that incorporate PILRA modifications alongside other relevant risk genes (e.g., APOE variants)

• Imaging innovations:

  • PET ligands targeting PILRA to enable non-invasive monitoring of expression in living subjects

  • Super-resolution microscopy techniques to visualize PILRA-ligand interactions at the nanoscale

  • Intravital microscopy approaches to monitor PILRA-expressing cells in real-time in vivo

• Computational and AI approaches:

  • Machine learning algorithms to predict PILRA-ligand interactions and antibody binding properties

  • Network analysis tools to better understand PILRA's position in immune regulatory networks

  • Systems biology approaches to model the effects of PILRA modulation across multiple biological scales

These innovations would significantly enhance our ability to understand PILRA biology and accelerate the development of therapeutic strategies targeting this receptor in various disease contexts .

How might PILRA research bridge fundamental immunology and clinical applications in precision medicine?

PILRA research represents a promising bridge between fundamental immunology and precision medicine applications:

• Genetic stratification for personalized therapeutics:

  • PILRA genotyping could identify individuals most likely to benefit from specific therapeutic approaches

  • The interaction between PILRA variants and other genetic factors (e.g., APOE in AD) suggests opportunities for combinatorial genetic profiling

  • Different PILRA-targeting strategies might be optimal depending on individual genetic backgrounds

• Biomarker development:

  • PILRA expression or activity measures could serve as biomarkers for disease progression or treatment response

  • In cancer, PILRA expression correlates with immunotherapy outcomes, suggesting utility in patient selection

  • Soluble PILRA levels might provide accessible biomarkers from blood or cerebrospinal fluid

• Multi-disease applications:

  • PILRA's emerging roles in both neurodegenerative diseases and cancer highlight its potential relevance across multiple conditions

  • Research findings in one disease context may inform applications in others

  • The connection with megaloblastic anemia suggests even broader clinical relevance

• Translational immunology insights:

  • PILRA research reveals fundamental principles about immune regulation that have clinical implications

  • Understanding how inhibitory receptors like PILRA balance immune activation informs broader immunomodulatory strategies

  • The pairing of PILRA with the activating receptor PILRβ exemplifies concepts relevant to immune checkpoint regulation

• Implementation science opportunities:

  • Development of accessible genetic testing for PILRA variants could enable population-level screening

  • Clinical decision support tools incorporating PILRA status alongside other factors could guide treatment selection

  • Health economics research could assess the value of PILRA-informed precision medicine approaches