INTERFERON-g PRODUCTION STIMULATED BY TWO PROTEINS PURIFIED FROM Bordetella pertussis
Wolf, P.S.; Lopes, A.P.Y.; Ferreira, V.R.F.; Cainelli Gebara, V.C.B.
Centro de Biotecnologia, Instituto Butantan , São Paulo, SP, Brazil.
We have been working on the immunomodulatory activities of a soluble fraction obtained from Bordetella pertussis. Two proteins (B1 and B2) were isolated from this soluble fraction, and were purified by anion-exchange chromatography. Our previous studies showed that both proteins are endowed with immunogenic and adjuvant properties as evaluated by antibodies production. In this work these proteins were used for the immunization of BALB/c mice mixed with a DPT (Diphtheria-Pertussis-Tetanus) vaccine formulated without aluminum hydroxide (NA-DPT). DPT vaccine produced at Instituto Butantan (DPTBut) was used as control. Spleen cells from immunized mice were evaluated for the production of Interferon-g (IFN-g) after in vitro stimulation with the vaccine antigens pertussis toxin (PT), diphtheria (DIF) and tetanus (TET) anatoxin. IFN-g concentration in the supernatants of cultured spleen cells was determined by ELISA.
The results showed that high levels of IFN-g were produced after stimulation with the vaccine antigens. Immunization with B1 alone or B1+NA-DPT induced an increased production of IFN-g after PT stimulation. A high production of IFN-g was observed after DIF stimulation of the cells from the group immunized with both proteins (B1+B2+NA-DPT). High levels of IFN-g were produced after TET stimulation and were observed in all groups. Studies are now in progress to evaluate other cytokines stimulated by the immunization with B1 and B2 proteins.
Supported by FAPESP, FUNDAÇÃO BUTANTAN.
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