Construction of CAR receptors to redirect T and NK cells to control invasive Candida albicans infection

Abstract

Invasive Candidiasis is an infection caused mainly by Candida albicans and is often associated with high rates of mortality. Candida infections affect 80% of the patients among all fungal infections in hospitals and, in Brazil, the incidence of Candidemia is approximately 2.5 cases in 1000 hospital admissions. Controlling Invasive Fungal Infections (IFIs) depends strongly on the innate immune response, such as NK cells, with subsequent involvement of the effector activity of T cells, mainly Th1 and Th17 subsets, and cytotoxic T cells. However, C. albicans can modify its morphology to evade the immune response by changing the expression of PAMPs in the cell wall. Thus, our proposal aims to redirect T and NK cells, through Chimeric Antigenic Receptors (CAR), for the specific recognition of C. albicans invasive form. First step will be to synthesize different CAR constructs with the scFv5, scFv12 or scFv k3-1 coding sequences. These synthesized sequences will be subcloned into a lentiviral backbone that contains the other portions of the CAR, and the lentiviral particles containing the CAR construct will be produced in HEK-293FT cells. Jurkat cells, NK-92 and PBMC will be transduced to assess the CAR specificity for C. albicans and the ability of CAR to induce the cytotoxic activity on C. albicans. The redirection of T cells via CAR will also be investigated in controlling the growth of C. albicans in infected NSG mice. The current proposal is very promising in the development of new therapies to control Invasive Candidiasis through T and NK cells expressing CAR. (AU)