THEMIS Antibody

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

THEMIS (Thymocyte-expressed molecule involved in selection), also known as Grb2-associating protein (Gasp), is a 72 kDa cytoplasmic phosphoprotein that plays a crucial role in T-cell development. THEMIS is essential for positive selection and the development of single-positive thymocytes, serving as a central regulator in late thymocyte development by controlling both positive and negative T-cell selection .

The protein's highest expression is found in CD4+CD8+ double-positive thymocytes, with lower expression in mature T cells . THEMIS functions by sustaining and integrating signals required for proper lineage commitment and maturation of T-cells, particularly through T-cell antigen receptor (TCR) signaling regulation .

This protein has attracted significant research interest because it represents a critical molecular component in understanding T-cell development and immune system regulation, with implications for autoimmune diseases and immunotherapies.

What types of THEMIS antibodies are currently available for research?

Several types of THEMIS antibodies are available for research applications:

These antibodies target different epitopes of THEMIS and vary in their species reactivity, with options available for detecting human, mouse, and rat THEMIS proteins .

What are the confirmed applications for THEMIS antibodies?

THEMIS antibodies have been validated for several research applications:

• Western Blotting (WB): Used to detect THEMIS protein in cell and tissue lysates. Most antibodies are optimized for this application, typically used at dilutions ranging from 1:100 to 1:1000 .

• Flow Cytometry: Particularly useful for detecting THEMIS in thymocytes and peripheral T cells. Monoclonal antibodies like 1TMYS are suitable for intracellular staining followed by flow cytometric analysis .

• Immunohistochemistry (IHC): Some antibodies have been validated for IHC-P (paraffin-embedded) applications, usually at dilutions of 1:100-500 .

• Immunofluorescence (IF): Several antibodies can be used for immunofluorescence applications, particularly for visualizing THEMIS localization in cells .

• ELISA: Selected antibodies have been validated for enzyme-linked immunosorbent assay applications .

Each application requires specific optimization and validation for reliable results, particularly regarding antibody concentration and sample preparation protocols.

How should I optimize intracellular staining for flow cytometric analysis of THEMIS?

For optimal intracellular staining of THEMIS for flow cytometry:

• Cell Preparation: Start with freshly isolated thymocytes or splenocytes. For in vitro studies, isolated cells should be rested for approximately 3 hours at 37°C before the assay to reduce background signaling .

• Fixation and Permeabilization: Use a dedicated intracellular fixation and permeabilization buffer set, such as the one referred to in the 1TMYS antibody protocol (Product # 88-8824-00). This is critical because THEMIS is a cytoplasmic protein .

• Antibody Titration: Carefully titrate the THEMIS antibody for optimal performance. The 1TMYS antibody, for example, can be used at ≤0.25 µg per test, where a test is defined as the amount of antibody that will stain a cell sample in a final volume of 100 µL .

• Cell Number Optimization: Cell numbers can range from 10^5 to 10^8 cells/test, but should be determined empirically for your specific experimental system .

• Protocol Selection: Follow specific protocols designed for cytoplasmic intracellular proteins, such as "Protocol A: Two step protocol for (cytoplasmic) intracellular proteins" .

• Controls: Include appropriate isotype controls to set quadrant markers for flow cytometric analysis, as demonstrated in the R&D Systems human THEMIS antibody testing .

• Co-staining: For T-cell subset analysis, consider co-staining with markers such as CD3, CD4, and CD8 to identify specific T-cell populations, as THEMIS expression varies across T-cell development stages .

What are the key considerations for detecting THEMIS by Western Blot?

When detecting THEMIS by Western Blot, researchers should consider:

• Sample Preparation:

  • Fresh tissue/cell lysates yield better results, particularly from thymocytes, splenocytes, or T-cell lines

  • Include phosphatase inhibitors if studying phosphorylated THEMIS

• Reduction Conditions:

  • Some THEMIS antibodies show differential recognition under reducing versus non-reducing conditions

  • The monoclonal antibody 719945 shows 100% cross-reactivity with recombinant human THEMIS under non-reducing conditions, but only approximately 10% cross-reactivity under reducing conditions

• Expected Molecular Weight:

  • THEMIS typically appears at approximately 72 kDa on SDS-PAGE

  • Be aware that post-translational modifications can alter migration patterns

• Dilution Optimization:

  • Start with manufacturer's recommended dilution (typically 1:1000 for polyclonal antibodies)

  • Perform dilution series to determine optimal signal-to-noise ratio

• Blocking Conditions:

  • BSA-based blocking solutions are often recommended, particularly for phospho-specific detection

  • 5% non-fat dry milk can be used for general THEMIS detection

• Detection Methods:

  • Enhanced chemiluminescence (ECL) is suitable for most applications

  • Fluorescence-based detection can provide better quantitation

• Controls:

  • Include positive controls such as mouse spleen or human K562 cells, which have been validated with available antibodies

  • Consider using THEMIS-knockout samples as negative controls when available

How can I validate the specificity of a THEMIS antibody?

Validating THEMIS antibody specificity is crucial for experimental reliability. Follow these approaches:

• Knockout/Knockdown Controls:

  • Use THEMIS-deficient samples (Themis^-/-^ mice tissues) as negative controls

  • siRNA or shRNA knockdown of THEMIS in cell lines provides partial reduction controls

  • Compare staining patterns between wild-type and Themis^-/-^ samples in the same experiment

• Blocking Peptides:

  • Pre-incubate the antibody with the immunizing peptide (many THEMIS antibodies specify their immunogen, e.g., "C-EERTINLPKSLKSH")

  • Compare the results with and without peptide blocking

• Multiple Antibody Validation:

  • Use antibodies targeting different epitopes of THEMIS (N-terminal vs. C-terminal)

  • Consistent results across different antibodies increase confidence in specificity

• Recombinant Protein Controls:

  • Test antibody against recombinant THEMIS protein

  • For example, E. coli-derived recombinant human THEMIS (Ala2-Pro282) has been used to validate antibodies

• Cross-Reactivity Testing:

  • If working across species, verify species cross-reactivity experimentally

  • Some antibodies show reactivity to human, mouse, and rat THEMIS due to sequence conservation

• Isoform Recognition:

  • Determine which THEMIS isoforms are recognized by your antibody

  • For instance, the 1TMYS antibody recognizes mouse THEMIS isoforms 1, 2, 3, and 4, but not isoform 5

• Mass Spectrometry Confirmation:

  • For definitive validation, perform immunoprecipitation followed by mass spectrometry to confirm the identity of the target protein

How can I study THEMIS phosphorylation dynamics in TCR signaling pathways?

Studying THEMIS phosphorylation requires specialized techniques due to its rapid phosphorylation following TCR stimulation:

• Stimulation Protocols:

  • Use tetramers or antibodies (anti-CD3/CD4) to stimulate thymocytes

  • Consider streptavidin crosslinking of anti-CD3/CD4 antibodies to mimic graded signal strengths that TCRs might generate in vivo

  • For physiological stimulation, use MHCp ligands with the OT-I TCR-transgenic model, which provides a full spectrum of signal strengths

• Timing Considerations:

  • THEMIS is quickly phosphorylated following TCR signaling, so include very early time points (15-30 seconds)

  • Design time-course experiments to capture the transient nature of phosphorylation events

• Detection Methods:

  • Use phospho-specific antibodies when available

  • Alternatively, perform immunoprecipitation with anti-THEMIS antibodies followed by western blotting with anti-phosphotyrosine antibodies (e.g., 4G10)

  • Consider using Phos-tag™ SDS-PAGE to detect mobility shifts caused by phosphorylation

• Co-Immunoprecipitation Studies:

  • THEMIS interacts with multiple signaling proteins including Grb2, PLC-γ1, and LAT

  • Use anti-THEMIS antibodies for immunoprecipitation followed by western blotting for associated proteins

  • The rabbit anti-Themis antibody (Millipore #06-1328) has been validated for immunoprecipitation in thymocytes

• Phosphatase Controls:

  • Include conditions with phosphatase inhibitors (positive control) and without (negative control)

  • Consider using phosphatase treatment of some samples to confirm phosphorylation-dependent effects

• Quantification:

  • Use densitometry to quantify phosphorylation levels

  • Normalize phospho-THEMIS signal to total THEMIS protein levels

• Downstream Signaling Analysis:

  • Monitor ERK phosphorylation and calcium mobilization as functional readouts of THEMIS-dependent signaling

  • Analyze the effect of THEMIS deficiency on SHP1 activation, which has been linked to THEMIS function in TCR signaling

What are the approaches to distinguish between THEMIS isoforms using antibodies?

Distinguishing between the five known isoforms of THEMIS requires careful antibody selection and complementary techniques:

• Epitope-Specific Antibody Selection:

  • Choose antibodies that target regions with sequence differences between isoforms

  • The 1TMYS monoclonal antibody recognizes isoforms 1, 2, 3, and 4, but not isoform 5 of mouse THEMIS

  • Consider using antibodies targeting different regions (N-terminal vs. C-terminal) for comparative analysis

• Western Blot Analysis:

  • Use high-resolution SDS-PAGE (8-10%) to separate isoforms based on molecular weight differences

  • Consider gradient gels for improved resolution of closely sized isoforms

  • Look for distinct banding patterns that correspond to different isoforms

• RT-PCR and qPCR Validation:

  • Design primers specific to unique regions of each isoform

  • Perform RT-PCR or qPCR to confirm expression of specific isoforms in your experimental system

  • This approach complements antibody-based detection methods

• Recombinant Protein Controls:

  • Express recombinant versions of each THEMIS isoform

  • Use these as positive controls to identify the migration pattern of each isoform on western blots

• Immunoprecipitation-Mass Spectrometry:

  • Immunoprecipitate THEMIS from your samples using a pan-THEMIS antibody

  • Analyze by mass spectrometry to identify peptides unique to specific isoforms

  • This approach provides definitive identification of expressed isoforms

• Functional Analysis:

  • Use isoform-specific knockdown or knockout approaches

  • Compare functional outcomes (e.g., T-cell development, TCR signaling) when specific isoforms are depleted

  • This approach helps determine the biological relevance of isoform differences

• Tissue and Developmental Stage Profiling:

  • Analyze expression patterns of THEMIS isoforms across different tissues and developmental stages

  • This may reveal context-specific roles for different isoforms

How can I analyze THEMIS-associated protein complexes in T cell signaling?

Analyzing THEMIS-associated protein complexes requires specialized techniques to preserve interactions:

• Optimized Co-Immunoprecipitation:

  • Use mild lysis conditions (e.g., 1% NP-40 or digitonin-based buffers) to preserve protein-protein interactions

  • Include phosphatase inhibitors to maintain phosphorylation-dependent interactions

  • Validate immunoprecipitation efficiency using anti-THEMIS antibodies suitable for IP, such as the rabbit anti-Themis antibody (Millipore #06-1328)

• Proximity Ligation Assays (PLA):

  • Use PLA to visualize and quantify THEMIS interactions with known partners (Grb2, PLC-γ1, LAT) in situ

  • This technique allows detection of protein interactions at endogenous expression levels within intact cells

• Mass Spectrometry Analysis:

  • Perform immunoprecipitation of THEMIS followed by mass spectrometry

  • Use both label-free and isotope labeling approaches (SILAC, TMT) for quantitative comparison

  • Compare resting vs. TCR-stimulated conditions to identify dynamic interaction partners

• Temporal Analysis of Complex Formation:

  • Design time-course experiments following TCR stimulation

  • THEMIS is quickly phosphorylated by SLP-76 following TCR signaling, suggesting rapid complex formation

  • Analyze samples at multiple time points (30 seconds to 30 minutes) to capture transient interactions

• Domain-Specific Interaction Mapping:

  • Use truncated or domain-specific THEMIS constructs to map interaction domains

  • This approach helps identify which regions of THEMIS mediate specific protein interactions

• Functional Validation of Interactions:

  • Use mutational analysis to disrupt specific interactions

  • Assess the functional consequences of disrupting THEMIS interactions on T-cell development and signaling

  • The established role of THEMIS in controlling both positive and negative T-cell selection provides functional readouts

• Analysis of THEMIS Role in Signal Modulation:

  • Examine how THEMIS affects the activation state of interacting proteins

  • THEMIS has been shown to attenuate TCR signal strength via SHP1 recruitment and activation in response to low-affinity TCR engagement

  • Analyze phosphorylation status of interaction partners in the presence and absence of THEMIS

What are common issues when using THEMIS antibodies and how can they be resolved?

Researchers often encounter several challenges when working with THEMIS antibodies:

• Low Signal in Western Blot:

  • Issue: Weak or absent bands when probing for THEMIS

  • Solutions:

    • Increase antibody concentration (try 1:500 instead of 1:1000)

    • Extend primary antibody incubation time (overnight at 4°C)

    • Use enhanced sensitivity detection systems

    • Ensure THEMIS is expressed in your sample (thymocytes show highest expression)

    • Check reduction conditions as some antibodies show differential recognition under reducing vs. non-reducing conditions

• High Background in Flow Cytometry:

  • Issue: Poor signal-to-noise ratio in flow cytometric analysis

  • Solutions:

    • Optimize fixation and permeabilization protocols specifically for intracellular staining

    • Include proper blocking steps (Fc block, serum matching host species)

    • Titrate antibody concentration carefully (≤0.25 µg per test is recommended for some antibodies)

    • Rest cells (3 hours at 37°C) before staining to reduce background activation

    • Use proper controls to set gates and compensation

• Cross-Reactivity and Specificity Issues:

  • Issue: Multiple bands or unexpected staining patterns

  • Solutions:

    • Validate with THEMIS-deficient samples when possible

    • Use antibodies targeting different epitopes to confirm specificity

    • Check if the antibody recognizes all or specific isoforms (e.g., 1TMYS recognizes isoforms 1-4, not 5)

    • Perform peptide blocking controls with the immunizing peptide

    • Confirm species reactivity (some antibodies are species-specific)

• Ineffective Immunoprecipitation:

  • Issue: Poor pulldown of THEMIS protein

  • Solutions:

    • Use antibodies specifically validated for IP applications

    • Optimize lysis conditions to maintain protein integrity

    • Increase antibody amount or lysate concentration

    • Extend incubation time with antibody

    • Consider using magnetic beads instead of agarose for improved recovery

• Variable Results Across Experiments:

  • Issue: Inconsistent detection of THEMIS across experiments

  • Solutions:

    • Standardize cell preparation and activation protocols

    • Monitor THEMIS expression levels, which vary during T-cell development

    • Use identical lot numbers of antibodies when possible

    • Include consistent positive controls in each experiment

    • Standardize lysate preparation and protein quantification methods

How can I optimize THEMIS detection in different T cell subsets and developmental stages?

Detecting THEMIS across T cell development requires consideration of its differential expression patterns:

• Sample Selection Based on Expression Levels:

  • CD4+CD8+ double-positive thymocytes show highest THEMIS expression levels

  • Single-positive thymocytes have intermediate expression

  • Mature peripheral T cells show lower expression levels

  • Plan experiments accordingly, with more sensitive detection methods for mature T cells

• Flow Cytometry Panel Design:

  • Include markers to identify specific T cell subsets (CD4, CD8, CD3)

  • Add markers for developmental stages (CD24, CD69, TCR levels)

  • Consider including intracellular phospho-specific markers to correlate THEMIS with signaling status

  • Use fluorophores with minimal spectral overlap with PE when using PE-conjugated THEMIS antibodies

• Cell Isolation Strategies:

  • For thymocyte subsets, consider magnetic or flow cytometric sorting of specific populations

  • When using OT-I TCR-transgenic models, Tap1^-/-^ background can be used to arrest development at the pre-selection DP stage

  • For peripheral T cells, use higher antibody concentrations due to lower THEMIS expression

• Fixation and Permeabilization Optimization:

  • Different T cell subsets may require adjusted fixation protocols

  • If analyzing both surface markers and intracellular THEMIS, optimize protocols to maintain surface epitope integrity

  • Consider using dedicated kits for detecting phosphorylated proteins if studying THEMIS phosphorylation status

• Activation-Dependent Detection:

  • THEMIS is quickly phosphorylated following TCR signaling

  • Include both resting and activated conditions when studying functional states

  • For activation, consider physiological stimuli like peptide-MHC complexes or antibody crosslinking

• Quantitative Analysis Approaches:

  • Use median fluorescence intensity (MFI) rather than percent positive for accurate quantification

  • When comparing across subsets, normalize THEMIS expression to appropriate housekeeping proteins

  • Consider analyzing THEMIS expression relative to TCR signal strength markers

• Imaging-Based Detection:

  • For tissue sections or sorted cells, optimize immunofluorescence protocols

  • Co-stain with markers of T cell subsets and signaling molecules

  • Consider confocal microscopy to analyze subcellular localization of THEMIS in different T cell populations

What are the best experimental approaches to study THEMIS function in T cell development?

Studying THEMIS function in T cell development requires integrated approaches:

• Genetic Models:

  • Themis^-/-^ mice show defective positive selection and fewer mature thymocytes

  • OT-I TCR-transgenic Themis^-/-^ mice provide a controlled system to study selection processes

  • Consider acute deletion of THEMIS using inducible Cre systems to distinguish developmental vs. maintenance roles

  • Bim knockout mice have been used in combination with THEMIS models to study apoptotic pathways

• Flow Cytometric Analysis of Development:

  • Quantify CD4/CD8 populations at different developmental stages

  • Assess positive and negative selection using markers like CD69, CD5, and annexin V

  • Combine with TCR signal strength markers (Nur77, CD5) to correlate with selection outcomes

  • Include intracellular staining for phosphorylated ERK1/2, which is critical for positive selection

• Calcium Signaling Assays:

  • Load thymocytes with Ca²⁺-sensitive dyes like Fluo-4

  • Co-label wild-type and Themis^-/-^ cells (e.g., with Cy5) to analyze them in the same sample

  • Stimulate with tetramers or antibodies while monitoring calcium flux

  • This approach reveals THEMIS's role in modulating TCR-induced calcium responses

• Biochemical Signaling Analysis:

  • Compare signaling in wild-type vs. Themis^-/-^ thymocytes

  • Focus on key pathways including:

    • ERK phosphorylation kinetics

    • PLC-γ1 phosphorylation

    • LAT phosphorylation

    • SHP1 activation

  • These analyses help elucidate THEMIS's role as a signal threshold regulator

• Ex Vivo Selection Assays:

  • Use peptide-loaded antigen-presenting cells with different-affinity peptides

  • Analyze selection outcomes in the presence and absence of THEMIS

  • This approach helps validate THEMIS's role as an "analog-to-digital converter" translating TCR affinity into selection outcomes

• Advanced Imaging Techniques:

  • Employ antigen-presenting lipid bilayers or peptide-loaded RMA-S cells for imaging

  • Monitor immunological synapse formation and dynamics

  • Track THEMIS localization during T cell-APC interactions

  • Correlate with signaling molecule recruitment and activation

• Integrated Multi-Parameter Analysis:

  • Combine multiple readouts (development, signaling, gene expression)

  • Consider single-cell approaches to account for cellular heterogeneity

  • Correlate THEMIS expression/function with T cell fate decisions

  • This comprehensive approach can reveal how THEMIS sets the signal threshold for positive and negative selection

What are the optimal storage and handling conditions for THEMIS antibodies?

Proper storage and handling of THEMIS antibodies is critical for maintaining their performance over time:

• Storage Temperature:

  • Long-term storage: -20°C to -70°C for most antibodies

  • Avoid storing at 4°C for extended periods, which can lead to degradation

  • Some antibodies contain 50% glycerol, allowing aliquoting without freeze/thaw cycles

• Aliquoting Recommendations:

  • Divide antibodies into single-use aliquots upon receipt

  • Typical aliquot volumes range from 10-50 µL depending on application

  • Use sterile tubes and conditions when preparing aliquots

• Freeze-Thaw Considerations:

  • Minimize freeze-thaw cycles as they can degrade antibody performance

  • Sources explicitly advise to "avoid repeated freeze-thaw cycles"

  • If using antibodies with 50% glycerol, aliquots may be taken without freeze/thawing

• Reconstitution Protocols:

  • For lyophilized antibodies, reconstitute in sterile PBS to a final concentration of 0.5 mg/mL

  • Allow complete dissolution before use or aliquoting

  • Filter through a 0.22 µm filter if sterility is required

• Buffer Compositions:

  • Many THEMIS antibodies are supplied in PBS with:

    • 50% glycerol (for stability)

    • 1 mg/ml BSA (as carrier protein)

    • 0.05% NaN₃ (as preservative)

  • Check specific product information as formulations vary between manufacturers

• Shelf-Life Guidelines:

  • Typical shelf life: 12 months from date of receipt at recommended storage temperature

  • After reconstitution: 1 month at 2-8°C or 6 months at -20 to -70°C under sterile conditions

  • Long-term stability (1+ year) is generally achieved at -20°C for antibodies in 50% glycerol

• Transportation Conditions:

  • Shipped on blue ice or dry ice depending on the formulation

  • Upon receipt, store immediately at recommended temperature

  • Inspect for any signs of degradation (precipitates, color changes)

How can researchers validate lot-to-lot consistency of THEMIS antibodies?

Ensuring lot-to-lot consistency is critical for reproducible research with THEMIS antibodies:

• Standard Validation Protocols:

  • Western blot using consistent positive control samples (mouse spleen, human K562 cells)

  • Flow cytometry on reference samples (thymocytes, peripheral blood lymphocytes)

  • Compare staining patterns, signal intensity, and background levels between lots

• Quantitative Performance Metrics:

  • Signal-to-noise ratio in Western blot or flow cytometry

  • Titration curves to determine effective working concentration

  • EC50 values from dilution series

  • Cross-reactivity profiles with related proteins

• Reference Standard Comparison:

  • Maintain a small amount of previous lot as reference standard

  • Run side-by-side comparison with new lot

  • Document relative performance across applications

• Epitope Mapping Confirmation:

  • Verify recognition of the intended epitope

  • For peptide antibodies, perform peptide blocking with the immunizing peptide

  • Confirm expected pattern of isoform recognition (e.g., 1TMYS antibody recognizes isoforms 1-4, not 5)

• Application-Specific Validation:

  • For Western blot: verify molecular weight, band pattern, and sensitivity

  • For flow cytometry: confirm population identification and staining intensity

  • For immunoprecipitation: assess pulldown efficiency and specificity

• Supplier Quality Documentation:

  • Request and review Certificate of Analysis from manufacturer

  • Many suppliers perform lot-specific testing (e.g., "Western blots performed on each lot")

  • Review lot-specific data provided by manufacturers

• Internal Reference Samples:

  • Maintain laboratory reference standards (cell lysates, fixed cells)

  • Document lot performance on these standards

  • Create a standardized scoring system for comparative evaluation

What are emerging applications of THEMIS antibodies in studying T cell signaling regulation?

THEMIS antibodies are finding new applications in advanced research contexts:

• Single-Cell Analysis of THEMIS Function:

  • Integration with mass cytometry (CyTOF) for multi-parameter analysis

  • Single-cell western blotting to analyze THEMIS expression heterogeneity

  • Correlation of THEMIS levels with signaling outcomes at single-cell resolution

• THEMIS in Immune Checkpoint Regulation:

  • Exploration of THEMIS's role in modulating T cell responses to checkpoint inhibitors

  • Analysis of THEMIS expression in tumor-infiltrating lymphocytes

  • Potential correlation with immunotherapy responsiveness

• THEMIS in Autoimmune Disease Mechanisms:

  • Given THEMIS's role in T cell selection, investigation of its dysregulation in autoimmunity

  • Analysis of THEMIS expression and function in patient-derived T cells

  • Potential therapeutic targeting based on its role in setting signal thresholds

• Integrated Phosphoproteomics:

  • Using THEMIS antibodies for immunoprecipitation followed by phosphoproteomics

  • Mapping the THEMIS-dependent phosphoproteome in T cells

  • Identifying novel THEMIS-regulated signaling pathways

• Super-Resolution Microscopy Applications:

  • Visualizing THEMIS within the nanoscale organization of signaling complexes

  • Analysis of THEMIS clustering and co-localization with TCR signaling components

  • Dynamic imaging of THEMIS recruitment to the immunological synapse

• CRISPR Screening with THEMIS Readouts:

  • Using THEMIS phosphorylation or localization as readouts in CRISPR screens

  • Identifying novel regulators of THEMIS function

  • Uncovering synthetic interactions with THEMIS in T cell development

• Computational Modeling of THEMIS Function:

  • Using quantitative data from THEMIS antibody-based experiments to inform mathematical models

  • Simulating THEMIS's role as an "analog-to-digital converter" in TCR signaling

  • Predicting T cell fate decisions based on THEMIS-dependent signaling parameters

How might THEMIS research contribute to understanding and treating immune disorders?

THEMIS research holds significant potential for translational applications:

• Biomarker Development:

  • THEMIS expression or phosphorylation status as potential biomarkers for T cell development disorders

  • Correlation of THEMIS function with immune dysregulation in patients

  • Potential diagnostic applications in primary immunodeficiencies

• Therapeutic Target Identification:

  • Given THEMIS's role in setting signaling thresholds, potential for therapeutic modulation in autoimmunity

  • Development of approaches to enhance or inhibit THEMIS function

  • Exploration of THEMIS-dependent pathways as alternative targets

• Understanding T Cell Selection Defects:

  • THEMIS's critical role in positive selection makes it relevant for understanding selection defects

  • Analysis of THEMIS expression and function in patients with T cell development abnormalities

  • Potential genetic screening for THEMIS variants in unexplained immunodeficiencies

• Enhancing Cancer Immunotherapy:

  • THEMIS modulation as a potential approach to enhance T cell anti-tumor responses

  • Analysis of THEMIS function in tumor-reactive T cells

  • Combining THEMIS targeting with existing immunotherapies

• Transplantation Immunology Applications:

  • Understanding how THEMIS influences alloreactive T cell responses

  • Potential for targeting THEMIS-dependent pathways to modulate graft rejection

  • Analysis of THEMIS function in regulatory T cells important for tolerance

• Vaccine Development Insights:

  • THEMIS's role in TCR signal modulation may inform approaches to enhance vaccine-induced T cell responses

  • Analysis of THEMIS function in memory T cell development

  • Potential adjuvant development targeting THEMIS-dependent pathways

• Precision Medicine Applications:

  • Stratification of patients based on THEMIS expression or function

  • Tailoring immunomodulatory therapies based on THEMIS-dependent signaling profiles

  • Development of companion diagnostics using THEMIS antibodies for immunotherapy selection