DENDRITIC CELL TARGETING IN CULTURED FISH IN ARGENTINA FOR VACCINE DEVELOPMENT.

GHERSA F.; QUATTROCCHI V; GAMMELLA M.; LANGELLOTTI A.; ZAMORANO P.; RAUQUE C.; PAPPALARDO J.

Mar del Plata

Congreso; Reunión Anual de Sociedades de Biociencia; 2019

Sociedad Argentina de Biociencia

Aquaculture is a fast-developing sector in the food industry worldwide. In Argentina two main species present an important economic value, these are Pacú (Piaractus mesopotamicus, Pm) and Rainbow Trout (Oncorhynchus mykiss, Om). Due to stress and changing environmental conditions cultured fish are exposed to, the use of antibiotics has become a common solution for treatment and avoidance of disease. This practice presents several problems such as overdose, contamination and resistance generation. The development of effective and affordable vaccines is necessary for aquaculture in order to produce safe products for consumption and the environment. This work focuses on the evaluation of a species unspecific nanovaccine platform in Pm and Om, composed of liposomes decorated with a1,2-mannobiose, a specific disaccharide that targets DC-SIGN receptor, mainly expressed on dendritic cells (DC). We cultured DC obtained from head kidney (HK), of Pm and Om in complete D-MEM (10 % FBS) for 1, 7 and 14 days at room temperature (RT) in order to obtain non-adherent cells, enriched in DC. These cells where later incubated for 30 m or 12 h at RT in D-MEM without FBS with undecorated liposomes for unspecific cell targeting (plain-L), a1,2-mannobiose decorated (Mana-L) and DOTAP (DOTAP-L) liposomes as a positive control, all marked with rhodamine. Prior liposome formulation and characterization with Z sizer was done. Incubation was stopped adding complete D-MEM. Cells were washed and fixed with PFA at 0.02 % w/v final concentration and then analyzed by flow cytometry. Results were statistically analyzed with two-way ANOVA followed by Bonferroni?s Test. Results demonstrate that HK cultures at day 7 and 14 are enriched in DC-SIGN expressing cells, and Mana-L targets specifically these cells (***p