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Journal of Radioanalytical and Nuclear Chemistry
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

Abstract  

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.

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Journal of Radioanalytical and Nuclear Chemistry
Authors:
J. Baptista
,
D. Vieira
,
A. Galisteo-Júnior
,
P. Caproni
,
M. Casare
,
H. de Andrade-Júnior
,
P. Spencer
, and
N. Nascimento

Abstract  

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.

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