IL17RB Mouse

What is the expression pattern of IL-17RB in different mouse tissues?

Mouse IL-17RB (also known as IL-17 Rh1, IL-17E R, and EVI27) shows a distinct tissue-specific expression pattern. Northern blot analysis reveals high expression in liver and testes, with lower expression levels detected in kidney and lung tissues . Additionally, IL-17RB is expressed in specific hematopoietic cell populations, including selected T cell, B cell, and myeloid cell lines . Recent research has identified significant IL-17RB expression in intestinal tuft cells and type 2 innate lymphoid cells (ILC2s) . The receptor's expression is notably upregulated under inflammatory conditions, suggesting a role in immune responses .

When investigating tissue-specific expression, researchers should consider using a combination of techniques:

• Northern blot analysis for tissue-level expression

• Flow cytometry for cellular-level expression

• Immunohistochemistry for spatial distribution within tissues

What is the molecular structure of mouse IL-17RB?

Mouse IL-17RB is a 499 amino acid type I membrane protein with distinct structural domains:

• Signal peptide: 17 amino acids

• Extracellular domain: 269 amino acids (Arg18-Gly286)

• Transmembrane domain: 21 amino acids

• Cytoplasmic tail: 192 amino acids

What detection methods are most effective for studying IL-17RB in mouse samples?

Multiple validated methods are available for IL-17RB detection in mouse samples:

Flow Cytometry:

• Anti-mouse IL-17RB monoclonal antibody (e.g., Clone #752101) has been validated for flow cytometric analysis of IL-17RB expression in transfected cell lines .

• Optimal for quantifying receptor expression at the cellular level with appropriate controls.

Western Blot:

• Goat anti-mouse IL-17RB polyclonal antibody can detect a specific band for IL-17RB at approximately 50 kDa in mouse liver lysates .

• Western blot analysis should be conducted under reducing conditions for optimal results.

Immunohistochemistry:

• IL-17RB can be detected in perfusion-fixed frozen sections using appropriate antibodies (3 μg/ml concentration) followed by HRP-conjugated secondary antibodies .

• Specific staining is typically localized to the cell surface.

When selecting a detection method, researchers should consider:

• The biological question being addressed

• Sample type and preparation requirements

• Quantitative vs. qualitative needs

• Available antibody specificity and validation data

How does IL-17RB participate in IL-25 signaling in mice?

IL-17RB serves as a critical component of the IL-25 signaling pathway in mice. Unlike other IL-17 family members, IL-25 requires a heteromeric receptor complex composed of both IL-17RB and IL-17RA for functional signaling . This has been conclusively demonstrated through several experimental approaches:

• Knockout mouse studies: Both IL-17RB KO and IL-17RA KO mice fail to produce IL-5 or IL-13 in response to IL-25 stimulation, unlike wild-type mice .

• Antagonistic antibody experiments: Treatment with antagonistic monoclonal antibodies to either IL-17RB or IL-17RA completely blocks IL-25-induced pulmonary inflammation and airway hyperresponsiveness in naive BALB/c mice .

• In vitro assays: Anti-mouse IL-17RB mAb and anti-mouse IL-17RA mAb significantly block IL-25-induced IL-5 production from wild-type BALB/c splenocytes .

Methodologically, researchers can study IL-17RB-mediated IL-25 signaling through:

• Cytokine production assays following IL-25 stimulation

• Phosphorylation studies of downstream signaling molecules

• Reporter assays for nuclear factor kappa-B activation

• Functional readouts in specific physiological contexts (e.g., pulmonary inflammation)

How does IL-17RB function differ between cellular compartments in mice?

IL-17RB function shows important cell type-specific roles in mice:

In ILC2s:

• IL-17RB expression is high in IL-5-expressing ILC2s .

• IL-17RB mediates direct stimulation of ILC2s by IL-25, leading to increased expression of IL-13 and Ki-67 .

• This response has been validated in Nmur1iCre;Il17rbfl/fl mice, which show near-complete Il17rb deletion in ILC2s and abrogated IL-13 and Ki-67 expression following succinate treatment .

In intestinal tuft cells:

• Tuft cells unusually co-express both IL-25 (ligand) and IL-17RB (receptor) .

• IL-17RB in tuft cells appears to restrain IL-25 bioavailability .

• Deletion of IL-17RB in intestinal epithelial cells (Vil1Cre;Il17rbfl/fl mice) results in:

  • Increased tuft cell numbers

  • Increased goblet cells

  • Elevated basal activity of the tuft cell-ILC2 circuit

  • Significantly increased IL-13 expression by small intestinal ILC2s

These findings suggest a regulatory role where tuft cell IL-17RB acts as a quenching mechanism for IL-25, restraining overactivation of ILC2s under homeostatic conditions.

What genetic models are available for studying IL-17RB function in mice?

Several genetic mouse models have been developed to study IL-17RB function:

Constitutive knockout models:

• Il17rb−/− mice: Complete absence of IL-17RB expression

• Il17ra−/− mice: Useful for studying the requirement of heteromeric receptor complexes

Conditional knockout models:

• Il17rbfl/fl mice: Enable tissue-specific deletion when crossed with appropriate Cre lines

• Vil1Cre;Il17rbfl/fl mice: Delete Il17rb specifically in intestinal epithelial cells

• Nmur1iCre;Il17rbfl/fl mice: Target IL-17RB deletion in ILC2s

• Il25iCre/+;Il17rbfl/fl mice: Delete IL-17RB in IL-25-expressing cells (primarily tuft cells)

Reporter mice:

• "YRS" mice expressing reporter alleles for ILC2 signature genes can be crossed with Il17rbfl mice to generate Il5R;Il17rbfl mice that allow visualization of IL-17RB expression in relation to IL-5-expressing cells

When selecting a genetic model, researchers should consider:

• The specific biological question

• Potential developmental compensations in constitutive knockouts

• Efficiency and specificity of Cre-mediated deletion

• Availability of appropriate controls (e.g., Cre-negative littermates)

How can researchers effectively study the IL-17RB-IL-25 axis in vivo?

To investigate the IL-17RB-IL-25 axis in vivo, researchers can employ several complementary approaches:

Intranasal IL-25 administration:

• Allows assessment of IL-25-induced pulmonary inflammation

• Parameters to evaluate include:

  • Bronchoalveolar lavage fluid eosinophil counts

  • Bronchoalveolar lavage fluid IL-5 and IL-13 concentrations

  • Goblet cell hyperplasia

  • Airway hyperresponsiveness

Blocking antibody administration:

• Anti-IL-17RB or anti-IL-17RA antibodies can block receptor function with temporal control

• Can be administered to wild-type mice or specific genetic models

• Example: Treatment of 2-week-old Vil1Cre;Il17rbfl/fl mice with anti-IL-17RB blocking antibody abolished the features of increased basal ILC2 activation

Succinate challenge models:

• Succinate activates tuft cells, triggering IL-25 release

• Useful for studying the tuft cell-IL-25-ILC2 circuit

• Parameter measurements should include ILC2 activation markers (IL-13, Ki-67)

Combined genetic and pharmacological approaches:

• Using conditional knockout models with additional interventions provides the most rigorous experimental design

• Example: Treating Il17rb conditional knockout mice with exogenous IL-25 to assess receptor specificity

What experimental considerations are important when analyzing IL-17RB-dependent phenotypes?

When analyzing IL-17RB-dependent phenotypes, researchers should consider:

Age-dependent effects:

• IL-25+ tuft cells emerge in the mouse small intestine shortly before weaning

• Different phenotypes may be observed in neonatal, juvenile, and adult mice

Compensatory mechanisms:

• Complete absence of IL-17RB may trigger compensatory upregulation of other signaling pathways

• Analysis of multiple timepoints following genetic deletion can help identify compensatory changes

Receptor complex considerations:

• IL-17RB forms a complex with IL-17RA

• Absence of IL-17RB may result in altered receptor chain pairing of IL-17RA

• Both receptor components should be analyzed when possible

Tissue-specific effects:

• IL-17RB function varies by tissue and cell type

• Comprehensive analysis should include multiple tissues (intestine, lung, etc.)

• Cell type-specific analyses are essential due to divergent roles in different cellular compartments

How can researchers overcome challenges in detecting IL-17RB protein expression?

Researchers may encounter several challenges when detecting IL-17RB:

Low expression levels:

• Use sensitive detection methods like flow cytometry or immunoprecipitation followed by Western blot

• Consider enriching for IL-17RB-expressing cell populations before analysis

• Use positive controls: mouse liver tissue shows reliable IL-17RB expression

• Avoid spleen tissue as a single source, as it may give negative results in some detection methods

Antibody specificity concerns:

• Validate antibodies using knockout tissues or cells

• Compare results from multiple antibody clones when possible

• For flow cytometry, always include isotype control antibodies

  • Example: When using rat anti-mouse IL-17RB (MAB10402), include rat IgG2A isotype control (MAB005)

Protein size variations:

• Expect IL-17RB to appear at approximately 50 kDa in Western blots under reducing conditions

• Alternative splicing may produce variants of different sizes

• Post-translational modifications may affect apparent molecular weight

What are the critical controls for IL-17RB functional studies in mice?

Rigorous controls are essential for IL-17RB functional studies:

Genetic model controls:

• Use littermate controls for conditional knockout models

• Include both Cre-negative (Il17rbfl/fl) and Cre-positive (Cre;Il17rb+/+) controls when possible

• Consider heterozygous models (Il17rb+/-) to evaluate gene dosage effects

Antibody blocking controls:

• Include isotype-matched control antibodies

• Titrate antibody concentrations to determine optimal blocking dose

• Perform parallel experiments with known IL-17RB ligands (IL-25) and non-ligands (IL-17A)

Stimulation controls:

• Include dose-response assessments for IL-25 stimulation

• Test specificity using related cytokines (IL-17B, which binds weakly to IL-17RB)

• Include positive controls for downstream readouts (e.g., direct stimulation of cytokine production)

Validation across multiple assay systems:

• Confirm key findings using both in vitro and in vivo approaches

• Verify results using different detection methodologies

• Assess functional outcomes at both molecular (signaling) and physiological (inflammation) levels

What are emerging areas of investigation for IL-17RB in mouse models?

Several cutting-edge research directions for IL-17RB in mice are emerging:

Single-cell analysis of IL-17RB-expressing populations:

• Application of single-cell RNA sequencing to identify heterogeneity within IL-17RB+ cell populations

• Trajectory analysis to understand developmental relationships

• Spatial transcriptomics to map IL-17RB expression within tissue microenvironments

IL-17RB in tuft cell biology:

• Further investigation of the unusual ligand-receptor co-expression in tuft cells

• Exploration of tuft cell IL-17RB as a potential therapeutic target

• Detailed mechanism of how IL-17RB restrains IL-25 bioavailability

IL-17RB in tissue-specific immune regulation:

• Role in tissue-specific allergic responses beyond the intestine and lung

• Function in tissue repair and remodeling processes

• Interaction with tissue-resident immune cells in different organ systems

IL-17RB in disease models:

• Investigation in models of allergic asthma and other Th2-associated pathologies

• Role in intestinal helminth infection responses

• Potential involvement in immune responses to tissue damage

How can new methodologies enhance IL-17RB research in mice?

Emerging methodologies that can advance IL-17RB research include:

CRISPR-Cas9 technologies:

• Generation of more precise genetic models with minimal off-target effects

• Introduction of specific point mutations to study structure-function relationships

• Development of CRISPR activation/inhibition systems for temporal control of IL-17RB expression

Advanced imaging approaches:

• In vivo imaging of IL-17RB-expressing cells using reporter mice

• Intravital microscopy to track IL-17RB+ cell dynamics

• Super-resolution microscopy to study IL-17RB clustering and localization

Systems biology approaches:

• Integration of transcriptomic, proteomic, and metabolomic data

• Network analysis of IL-17RB signaling pathways

• Computational modeling of IL-25/IL-17RB signaling kinetics

Organoid models:

• Intestinal organoids to study the tuft cell-ILC2 circuit ex vivo

• Co-culture systems with epithelial and immune components

• Patient-derived organoids for translational research