Authors:J. Baptista, D. Vieira, A. Galisteo-Júnior, P. Caproni, M. Casare, H. de Andrade-Júnior, P. Spencer, and N. Nascimento
We investigated the immunological behavior of BTHX-1, before and after irradiation. SDS-PAGE showed that BTHX-1 irradiated
in the presence of NaNO3, had its structure preserved. Animals’ plasma immunized with native BTHX-1 had high IgG1 titers. The irradiated protein induced
high titers of IgG2b. When the toxin was irradiated with t-butanol, there was a slight decrease in the production of IgG2b.
Real-time PCR showed that both the IL-2 as for IL4 was more expression from the cells of the animals immunized with BTHX-1
irradiated. These results indicate that irradiation of proteins leads to significant structural modifications.
Authors:J. Baptista, D. Vieira, A. Galisteo Júnior, O. Higa, M. Casare, C. Yonamine, P. Caproni, L. Campos, H. de Andrade Júnior, P. Spencer, and N. Nascimento
In this work, the authors investigated the immunological behavior of bothropstoxin-I (BTHX-1), before and after irradiation
process, and also the influence of scavengers substances on protein alterations induced by free radical production. Structural
modifications were investigated by SDS-PAGE in reducing or non-reducing conditions. In vitro cytotoxicity assay was performed
to test average toxic activities of BTHX-I. BALB/c Isogenic mice were immunized with irradiated or non-irradiated (native)
forms of BTHX-I and antibody titers and isotypes were determined by ELISA method. Expression of murine cytokines was analyzed
by using expression data obtained by quantitative real-time PCR (qPCR) assays. The results indicate that irradiation of proteins
leads to significant structural modifications, and also changes the cytokines profile during immunization process, regarding
a suitable approach to new immunogenic production.