Objective. To determine whether nanoparticles (NP) made from FDA-approved biopolymers could be incorporated into a novel clinically translatable dendritic cell (DC)-based therapy for HNSCC. Methods. Head and neck squamous cell carcinoma lines were used to make lysates employing multiple techniques, and with varying amounts of protein so as to improve encapsulation within differing molecular weights of Poly (lactic-co-glycolic acid) (PLGA) polymers. NP were fabricated by a double emulsion solvent-evaporation technique with flash freeze-lyophilization. Encapsulation and release profile were studied to determine the ideal formulation. These findings were adopted for encapsulating tumour material from five patients with advanced HNSCC. Pattern of protein release was detected using Silver stained SDS-PAGE gels and immunoblotting for p53. GM-CSF/IL-4 DC made from blood monocyte precursors were loaded with either the soluble or encapsulated versions of tumour lysate, and used for stimulation of magnetic bead-isolated autologous CD8+ T-cells. Differences in Th1/Th2 responses were estimated by Cytometric bead array (CBA). NP morphology and DC loading were evaluated by scanning electron microscopy. Results. Optimal release was recorded from 80 K PLGA NP containing a lysate made using four cycles of freeze-thaw plus 15-s sonication followed by brief lyophilization. NP were small enough to be phagocytosed by DC. Encapsulation and release was directly proportional to the lysate concentration with no major antigen degradation. In four patients, CBA showed significantly increased IFN-gamma or decreased IL-10 (P = 0.012) production associated with NP encapsulation. Conclusions. Biopolymer NP can efficiently deliver the panoply of unknown tumour antigens. This strategy has been optimized for use in a clinical trial. References. 1 Sandor M., Enscore D., Weston P et al. (2001) Effect of protein molecular weight on release from micron-sized PLGA microspheres. J. Control. Release 76, 297-311 2 Elamanchili P., Lutsiak C., Hamdy S et al. (2007) 'Pathogen-mimicking' nanoparticles for vaccine delivery to dendritic cells. J. Immunother.30, 378-395.
|Translated title of the contribution||Biopolymer nanoparticles as antigen delivery vehicles for immunotherapy of head and neck squamous cell carcinoma (HNSCC)|
|Pages (from-to)||304 - 304|
|Number of pages||1|
|Publication status||Published - Jun 2008|