Lázaro-Gorines, RodrigoLópez-Rodríguez, Juan CarlosBenedé, SaraGonzález, MiguelMayorga, CristobalinaVogel, LotharMartínez-Del-Pozo, ÁlvaroLacadena, JavierVillalba, Mayte2024-02-122024-02-122020-07-23http://hdl.handle.net/10668/4059http://hdl.handle.net/20.500.12105/18081Immunotoxins appear as promising therapeutic molecules, alternative to allergen-specific-immunotherapy. In this work, we achieved the development of a protein chimera able to promote specific cell death on effector cells involved in the allergic reaction. Der p 1 allergen was chosen as cell-targeting domain and the powerful ribotoxin α-sarcin as the toxic moiety. The resultant construction, named proDerp1αS, was produced and purified from the yeast Pichia pastoris. Der p 1-protease activity and α-sarcin ribonucleolytic action were effectively conserved in proDerp1αS. Immunotoxin impact was assayed by using effector cells sensitized with house dust mite-allergic sera. Cell degranulation and death, triggered by proDerp1αS, was exclusively observed on Der p 1 sera sensitized-humRBL-2H3 cells, but not when treated with non-allergic sera. Most notably, equivalent IgE-binding and degranulation were observed with both proDerp1αS construct and native Der p 1 when using purified basophils from sensitized patients. However, proDerp1αS did not cause any cytotoxic effect on these cells, apparently due to its lack of internalization after their surface IgE-binding, showing the complex in vivo panorama governing allergic reactions. In conclusion, herein we present proDerp1αS as a proof of concept for a potential and alternative new designs of therapeutic tools for allergies. Development of new, and more specific, second-generation of immunotoxins following proDerp1αS, is further discussed.engVoRhttp://creativecommons.org/licenses/by/4.0/AllergyDer p 1ImmunotoxinsBasophilsCell deathPyroglyphidaeHipersensibilidadInmunotoxinasBasófilosMuerte celularAllergensAnimalsAntigens, DermatophagoidesArthropod ProteinsBasophilsCell DegranulationCell LineCells, CulturedCysteine EndopeptidasesDermatophagoides pteronyssinusDesensitization, ImmunologicHumansHypersensitivityImmunoglobulin EImmunotoxinsRecombinant ProteinsCell DegranulationCell DeathAttribution 4.0 International3270397210.1038/s41598-020-69166-w2045-2322Scientific Reportsopen access