Pan-allergens can contribute to severity and perpetuation of allergic diseases, as a result of leading to IgE-medicated poly-sensitization and cross-reactivity to human self-antigens. Cyclophilins are well-known pan-allergens of various groups of cross-reactive allergens. CyPs have been identified with five classic CyP isoforms (CyPA, B, C, D and E). In the present study, the physicochemical and immunologic characteristics of CyPs were predict by bioinformatics and immunoinformatics approaches. The results indicated that CyPA family possesses multiple identified allergens. CyPE showed closer evolutionary relationship and high sequence identity with allergenic CyPA and the structure and antigenicity of CyPE were conserved significantly compared with allergenic CyPA. To verify results of In silico analysis，CyPE from HMD were successfully constructed and purified. ELISA resuls indicated that 2/14 patient serum had positive reactions to CyPE. CyPE is low antigenicity but sufficient to induced significant airway inflammation in mouse models and IgE, IgG1, eosinophil infiltration, IL-4 and co-stimulatory molecules of CyPE were markedly lower levels than Der f 29. Structure and immunoinformatics analysis demonstrated that RNA-binding motif of CyPE may reduce the immunogenicity of CyPE and structure variation of B-cell epitopes is the reason why CyPB is no IgE activity. This work is of significance in the diagnosis and treatment of allergy.