Conjugation of a TLR5 agonist to the Plasmodium falciparum Circumsporozoite (CS) protein enhances murine and human Dendritic cells (DC) interaction in vitro and increases protective immunity following intranasal immunization in mice (37.4)

Abstract

Abstract A needle-free malaria vaccine has been developed based on intranasal (I.N) immunization with a recombinant P. falciparum circumsporozoite (CS) protein conjugated to the TLR5 agonist flagellin. Mice immunized I.N, but not subcutaneously, with flagellin-modified CS developed sporozoite neutralizing antibodies that protect against challenge with a transgenic rodent parasite expressing P. falciparum CS repeats. To better understand the role of innate immune cells in the induction of protective immunity, we analyzed the interaction of flagellin-modified CS with murine bone marrow-derived dendritic cells (DC) and human monocyte-derived DC in vitro. The fusion protein was rapidly taken up by DC and remained detectable in the cytosol as long as 24 hours post antigen pulse. A murine DC line (D1), and a sub-population of human DC, matured and expressed increased levels of CD40 and CD86 following stimulation with flagellin-modified CS. In I.N. immunized mice, the Nasopharyngeal-associated Lymphoid Tissue (NALT) was enlarged 2-fold over naïve NALT, and increased levels of CD11c+ DC were observed by confocal microscopy and FACS analysis. NALT cell subpopulations exhibited a higher B:T cell ratio than those observed in lymph nodes. A better understanding of innate and adaptive immune responses elicited in NALT by TLR ligand-modified CS protein will help identify critical immune parameters required for the induction of sporozoite specific immunity that protects from malaria infection.