Autor: |
Martinez-Peinado N; Barcelona Institute for Global Health (ISGlobal), Hospital Clínic-University of Barcelona, 08036 Barcelona, Spain.; Secció de Parasitologia, Departament de Biologia, Sanitat i Medi Ambient, Facultat de Farmàcia i Ciències de l'Alimentació, Universitat de Barcelona, 08007 Barcelona, Spain., Gabaldon-Figueira JC; Barcelona Institute for Global Health (ISGlobal), Hospital Clínic-University of Barcelona, 08036 Barcelona, Spain., Martinez-Añon I; Barcelona Institute for Global Health (ISGlobal), Hospital Clínic-University of Barcelona, 08036 Barcelona, Spain., Rodríguez-Gordo C; Laboratori de Referència Catalunya, 08820 Barcelona, Spain., Robleda-Castillo R; Barcelona Institute for Global Health (ISGlobal), Hospital Clínic-University of Barcelona, 08036 Barcelona, Spain., Pinazo MJ; Barcelona Institute for Global Health (ISGlobal), Hospital Clínic-University of Barcelona, 08036 Barcelona, Spain.; Drugs for Neglected Diseases initiative (DNDi), 1202 Geneva, Switzerland.; CIBER de Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC, ISCIII), 28220 Madrid, Spain., Bigey P; Université Paris Cité, CNRS, INSERM, UTCBS, F-75006 Paris, France.; Chimie ParisTech, PSL University, F-75005 Paris, France., Gascon J; Barcelona Institute for Global Health (ISGlobal), Hospital Clínic-University of Barcelona, 08036 Barcelona, Spain.; CIBER de Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC, ISCIII), 28220 Madrid, Spain., Alonso-Padilla J; Barcelona Institute for Global Health (ISGlobal), Hospital Clínic-University of Barcelona, 08036 Barcelona, Spain.; CIBER de Enfermedades Infecciosas, Instituto de Salud Carlos III (CIBERINFEC, ISCIII), 28220 Madrid, Spain. |
Abstrakt: |
Mounting a balanced and robust humoral immune response is of utmost importance for reducing the infectivity of Trypanosoma cruzi . While the role of such a response in controlling the infection is well known, there is a lack of tools that can be used to quickly evaluate it. We developed a serum parasite inhibition assay (to evaluate changes in the parasite infection after exposing infective T. cruzi trypomastigotes to serum samples from infected patients). It is based on Vero cells as the hosts and the Tulahuen β-galactosidase parasite strain, genetically engineered to be quantifiable by spectrophotometry. In parallel, we developed an in-house ELISA to correlate the anti- T. cruzi antibody titres of the clinical samples with their observed anti-parasitic effect in the serum parasite inhibition assay. Serum samples from chronically T. cruzi -infected patients significantly inhibited parasite invasion in a titre-dependant manner, regardless of the patient's clinical status, compared to samples from the non-infected controls. In addition, there was a clear correlation between the reactivity of the samples to the whole-parasite lysates by ELISA and the inhibitory effect. The results of this work confirm the previously described anti-parasitic effect of the serum of individuals exposed to T. cruzi and present a framework for its large-scale evaluation in further studies. The serum parasite inhibition assay represents a reproducible way to evaluate the intensity and anti-parasitic effect of humoral responses against T. cruzi , which could be applied to the evaluation of candidate antigens/epitopes in the design of Chagas disease vaccine candidates. |