Highly tailorable gellan gum nanoparticles as a platform for the development of T cell activator systems

Abstract

Background: T cell priming has been shown to be a powerful immunotherapeutic approach for cancer treatment in terms of efcacy and relatively weak side efects. Systems that optimize the stimulation of T cells to improve therapeutic efcacy are therefore in constant demand. A way to achieve this is through artifcial antigen presenting cells that are complexes between vehicles and key molecules that target relevant T cell subpopulations, eliciting antigenspecifc T cell priming. In such T cell activator systems, the vehicles chosen to deliver and present the key molecules to the targeted cell populations are of extreme importance. In this work, a new platform for the creation of T cell activator systems based on highly tailorable nanoparticles made from the natural polymer gellan gum (GG) was developed and validated. Methods: GG nanoparticles were produced by a water in oil emulsion procedure, and characterized by dynamic light scattering, high resolution scanning electronic microscopy and water uptake. Their biocompatibility with cultured cells was assessed by a metabolic activity assay. Surface functionalization was performed with anti-CD3/ CD28 antibodies via EDC/NHS or NeutrAvidin/Biotin linkage. Functionalized particles were tested for their capacity to stimulate CD4+ T cells and trigger T cell cytotoxic responses. Results: Nanoparticles were approximately 150 nm in size, with a stable structure and no detectable cytotoxicity. Water uptake originated a weight gain of up to 3200%. The functional antibodies did efciently bind to the nanoparticles, as confrmed by SDS-PAGE, which then targeted the desired CD4+ populations, as confrmed by confocal microscopy. The developed system presented a more sustained T cell activation over time when compared to commercial alternatives. Concurrently, the expression of higher levels of key cytotoxic pathway molecules granzyme B/perforin was induced, suggesting a greater cytotoxic potential for future application in adoptive cancer therapy. Conclusions: Our results show that GG nanoparticles were successfully used as a highly tailorable T cell activator system platform capable of T cell expansion and re-education.