| Přispěvatelé: |
[Lázaro Gorines,R, López Rodríguez,JC, Benedé,S, Martínez del Pozo,A, Lacadena,J, Villalba,M] Biochemistry and Molecular Biology Department, Chemical Sciences Faculty, Complutense University of Madrid, Madrid, Spain. [González,M, Mayorga,C] Allergy Research Laboratory, IBIMA, Hospital Regional Universitario de Málaga, UMA, Málaga, Spain. [Mayorga,C] U.G.C. Allergy, IBIMA, Hospital Regional Universitario de Málaga, UMA, Málaga, Spain. [Vogel,L] Division of Allergology, Paul-Ehrlich-Institut, Langen, Germany., Tis work was supported by the Universidad Complutense de Madrid under Grants (PR75/18-21563, PR87/19-22627), SAF2017-86483-R to M.V from the Ministerio de Economía y Competitividad. The Thematic Networks and Co-operative Research Centres: ARADyAL (RD16/0006/0014, RD16/0006/0001) from the Instituto de Salud Carlos III (ISCIII). R.L.G. was recipient of a predoctoral Research contract. Te FPU predoctoral contract to J.C.L.R. is supported by the Spanish Ministerio de Educación, Cultura y Deporte. |
| Popis: |
Immunotoxins 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. |
