Vaccine against Bovine Herpesvirus 1 using the Fragment C of the Tetanic Toxin as an adjuvant and an attenuated Salmonella Strain to deliver it.

IGNACIO CEBRIAN; SEBASTIÁN PAPPALARDO; CECILIA LANGELLOTTI; OSCAR TABOGA; ALEXANDER BATISTA DUHARTE; JOSÉ CHABALGOITY; ANA SADIR; PATRICIA ZAMORANO

Varadero, Cuba.

Workshop; 3rd International Workshop on Vaccine Adjuvants and GlycoConjugates; 2006

Cuban Society of Immunology (CSI) y Latin American Association of Immunology (ALAI)

Bovine Herpesvirus 1 (BHV-1) is a pathogen that affects cattle causing many diseases and important economic losses. In countries with high prevalency like Argentina, vaccination is the main strategy. Intending to make an effective vaccine we have developed a live system to release heterologous antigens using an attenuated Salmonella strain. These attenuated bacteria replicate for several weeks and induce an infection that the individual can support. But while the bacteria persist stimulate the humoral and cellular immune response for the Salmonellas´ antigens and for the heterologous antigens as well. These antigens are expressed and presented on the APC cells directly. We have transformed Salmonella typhimurium LVR 03 with the plasmid pTECH2 wich express an immunogenic peptide (from gD) repeated in tandem. In our laboratory was cloned this region of the gD protein of BHV-1 once, twice and four times. Fused with the foreign antigen it is the C fragment of the tetanic toxin (TetC) wich would supply adjuvant properties. Multiple cloning was confirmed by restriction enzymes analysis and DNA sequencing. The expression of the fusion protein was detected with Western Blot analysis. The different vaccine combinations were tested on BALB/c mices and via three inoculation routes (oral, intranasal and intravenous). Serum samples were analyzed by ELISA, Western Blot and Immunofluorescence. The obtained results indicate that the system used to release antigens in our experiments is working properly. Specific humoral immune response against Herpesvirus gD is observed, and it is highly dependent on the vaccine combination used and on the inoculation route administred. These encouraging results allow us to move on the bovine experimental model, wich represents the main target of our investigation project.