Background <p>Peptide immunotherapy (PIT) offers a safe and effective treatment with minimal side effects. This study aims to identify B-cell epitopes of a novel allergen from <i>Blomia tropicalis</i> (<i>B. tropicalis</i>), specifically the Chitin-binding domain type 2 (ChtBD2) protein, and evaluate the therapeutic effects of peptide treatment in a murine model.</p> Methods <p>Using Alphafold2, the 3D structure of ChtBD2 was constructed. AI-based and traditional computational tools predicted the predominant B-cell epitopes. Twelve synthesized peptides were assessed for allergenicity and immunogenicity. A murine model of <i>B. tropicalis</i>-induced allergic airway inflammation mimicking human atopic asthma was developed and analyzed.</p> Results <p>Predominant B-cell epitopes of ChtBD2 were identified as promising IgE-binding domains. Peptide 1 (PT1: 1–15) showed significant IgE-binding activity and the highest inhibition rate in competitive IgE-binding assays. PT1 upregulated IL-4, IL-13, and CD63 in <i>B. tropicalis</i>-sensitized patients’ PBMCs and basophils, respectively. Notably, IT groups showed reduced lung cellular infiltration and type 2 cytokine expression in BALF. Specific IgE levels were reduced, with a decline in the IgG₁/IgG_2a ratio.</p> Conclusions <p>This study represents the first AI-facilitated development of a B-cell epitope-based ChtBD2 PIT, showing promise as an immunotherapy for <i>B. tropicalis</i>-allergic patients with reduced allergenicity and high immunogenicity in inducing IgG-blocking antibodies.</p> Clinical trial <p>Not applicable.</p>