DEVELOPMENT OF A NOVEL MULTI-EPITOPE ANTIGEN EFFECTIVE TO CONTROL TRYPANOSOMA CRUZI INFECTION

VÁZQUEZ, MARÍA E; ZABALA, BRENDA A; MESÍAS, ANDREA C; PARODI, CECILIA; PÉREZ BRANDÁN, CECILIA; ACUÑA, LEONARDO

Virtual

Congreso; Congreso conjunto SAIB-SAMIGE 2021; 2021

SAIB- SAMIGE

DEVELOPMENT OF A NOVEL MULTI-EPITOPE ANTIGEN EFFECTIVE TO CONTROL TRYPANOSOMA CRUZI INFECTIONMaría Elisa Vázquez1, Brenda Zabala1, Andrea C. Mesías1, Cecilia Parodi1, Cecilia Pérez Brandán 1, Leonardo Acuña 1.1 IPE-CONICET, Salta, Argentina.E-mail: mariaelisavzqz@gmail.comChagas disease (CD) is a neglected disease caused by a flagellar protozoon named Trypanosoma cruzi, that affect over 8 million people around the world. CD is an endemic problem in Latin American and the principal cause of an infectious heart disease in the world. Nowadays, there is not an available vaccine for the prevention of this silent illness, and the research around vaccine development has not yet reach a complete protection against the parasite. For this reason, in recent years innovative approaches were studied for advancement in this field. One of these perspectives is the multi-component vaccine strategy, that mimic the natural infection in a better fashion. In this manner, we constructed a chimeric fusion protein based on T. cruzi antigens. For the construction, two fragments of antigens of T. cruzi were used. On one hand, the N-terminus Tc52 (N-Tc52) is a protein that develops a robust humoral response, and in the other hand TSKB20 is an epitope of TS protein that possess immunodominance in cellular response against the parasite. N-Tc52 was amplified by PCR from T. cruzi CL Brener strain and subsequently reamplified to incorporate, with specific primers, two TSKB20 sequences in tandem. Next, this genetic construct was cloned into a bacterial plasmid, pRSET-A and was sequenced, and indeed, confirmed no mutation. Finally, we expressed and purified the chimeric protein resulting (N-Tc52/TSKB20) and its primary structure was confirmed by mass spectrometry. An immunization scheme in mice was diagrammed to prove the biological activity of N-Tc52/TSKB20. Animals were inoculated with chimera plus an adjuvant saponin type 3 times separated between 21 days, controls were added too. Blood was collected before each dose and 21 days after last dose, the half of animals were sacrificed for measure the immune response and the other half was challenged with a lethal dose of T. cruzi trypomastigotes. Parasitemias were recorded twice a week for 25 days for tested vaccine efficiency and then animals were sacrificed. Samples were taken to analyze the expansion of immune response. In brief, mice inoculated with chimeric protein were able to control parasitemias and exhibited an immune response against T. cruzi in comparation with controls.