CCR3 Antibody

Here’s a structured FAQ collection for CCR3 antibody research, synthesized from peer-reviewed studies and patents:

How to validate CCR3 antibody specificity in experimental models?

Methodology:

• Use Western blotting with CCR3-transfected cell lysates to confirm target binding (e.g., studies demonstrated specificity using HEK293 cells expressing CCR3) .

• Employ flow cytometry with CCR3-knockout cells or competitive binding assays using CCR3 ligands (e.g., CCL11/eotaxin) .

• Validate via immunohistochemistry in tissues with known CCR3 expression (e.g., eosinophil-rich lung sections in allergic asthma models) .

What are optimal animal models for studying CCR3 antibody therapeutic effects?

Approach:

• Ovalbumin (OVA)-sensitized mice: Standard for allergic asthma/rhinitis, enabling assessment of eosinophil infiltration, mucus hypersecretion, and cytokine profiles .

• Key endpoints:

  Parameter	Measurement Technique	Example Outcome (CCR3 mAb vs. Control)
  Eosinophil count	Bronchoalveolar lavage (BAL)	↓ 50–70%
  Cytokine levels	ELISA (IL-4, IL-5, IL-13)	↓ Th2 cytokines by 30–60%
  Airway remodeling	Histopathology (H&E staining)	Reduced goblet cell hyperplasia

How to address variability in CCR3 antibody efficacy across studies?

Troubleshooting:

• Dose optimization: Test multiple concentrations (e.g., 5–20 µL/mg in murine models) .

• Administration route: Compare intraperitoneal (systemic) vs. intranasal (localized) delivery .

• Model heterogeneity: Standardize allergen exposure protocols (e.g., OVA challenge duration, adjuvant use) .

How does CCR3 antibody epitope specificity influence functional outcomes?

Epitope Mapping Insights:

• Critical residues: Alanine scanning revealed Ala2, Phe3, Asn4, Thr5 in the N-terminus are essential for antibody binding .

• Functional implications:

  Antibody Clone	Epitope Region	Impact on CCR3 Function
  C3Mab-3	N-terminal	Blocks ligand binding (CCL11/CCL5)
  C3Mab-4	N-terminal	Partial inhibition of chemotaxis
  J073E5	N-terminal	Reduces eosinophil migration by 80%

How to resolve contradictions in cytokine modulation data post-CCR3 antibody treatment?

Analysis Framework:

• Context-dependent effects:

  • In OVA models, CCR3 blockade reduces IL-4/IL-5 (Th2 cytokines) but may spare IFN-γ (Th1) .

  • Tissue-specific outcomes: Nasal mucosa shows stronger anti-inflammatory effects than lung tissue .

• Mechanistic overlap: CCR3 antibodies may concurrently upregulate regulatory T-cells (Tregs), complicating cytokine profiles .

What combinatorial strategies enhance CCR3 antibody efficacy?

Synergistic Approaches:

• Adjuvant immunotherapy: Co-administer CpG oligonucleotides to shift Th2/Th1 balance .

• Dual cytokine targeting: Pair with anti-IL-5 antibodies to amplify eosinophil suppression .

• Dosage schedule:

  Combination Partner	Timing Relative to CCR3 mAb	Outcome Improvement
  Dexamethasone	24 hours post-mAb	↓ Mucus production
  Anti-IL-4Rα	Concurrent administration	↓ IL-13 by 40%

How to control for CCR3 polymorphism effects in translational studies?

Best Practices:

• Use species-matched antibodies: Murine CCR3 mAbs (e.g., C3Mab-3) show <10% cross-reactivity with human CCR3 .

• Validate in primary cells: Test eosinophils from atopic donors to confirm clinical relevance .

What metrics distinguish CCR3 antibody-mediated immune modulation from nonspecific effects?

Key Metrics:

• Eotaxin-1 (CCL11) neutralization: Measure reduced chemotaxis in transwell assays (≥60% inhibition) .

• T-cell polarization: Assess Th2→Treg shifts via flow cytometry (e.g., FoxP3+ CD4+ T-cells) .