The fusion protein BLS-FliC has the capacity to triggers dendritic cells activation in a TLR4 and TLR5 dependent way

ERREA AGUSTINA; GRISELDA MORENO; ROSSI ANDRES H.; HIRIART YANINA; SIRARD, JEAN CLAUDE; GOLDBAUM, FERNANDO; BERGER, PAULA; RUMBO MARTIN

Los Cocos

Congreso; LXI Reunión Anual de la Sociedad Argentina de Inmunología; 2013

Sociedad Argentina de Inmunología

There has been increasing research in the field of subunit vaccines that highlighted the need for adjuvant development. TLR agonists are adjuvant candidates based on their capacity to boost adaptive responses..We have generated a fusion protein combining Lumazine synthase from Brucella spp. (BLS), which has shown TLR4 dependent properties, and flagellin (FliC), a TLR5 agonist. In the present work, we addressed the potential of the fusion protein BLS-FliC to induce the activation of dendritic cells (DC), an essential step for the induction of the adaptive response. We also studied the contribution of TLR5, TLR4 and MyD88 in BLS-FliC induced DC activation. Bone marrow dendritic cells (BMDC) were derived from C57BL/6 mice (wt), TLR4-defficient, TLR5-/- or MyD88-/- mice with GM-CSF and stimulated 18hs with BLS-FliC. Response was compared with the one elicited by stimulation with FliC and of BLS. LPS-stimulated and non treated cells were used as positive and negative controls. BMDC activation was determined measuring the levels of co-stimulatory molecules by flow citometry and cytokine secretion by ELISA. BLS-FliC induced increases in the expression of CD80 and CD86 and the secretion of IL12p40 and IL-6 on wt BMDC (p< 0.001 vs non treated cells). Moreover, at the employed doses, all measured parameters were higher for BLS-FliC treatment compared to simulation with BLS or FliC as single molecules (p