Chemical synthesis and immunological evaluation of cancer vaccines based on ganglioside antigens and α-galactosylceramideElectronic supplementary information (ESI) available. See DOI: https://doi.org/10.1039/d4md00387j

iNKT cells – often referred as the “Swiss Army knife” of the immune system – have emerged as central players in cancer vaccine therapies. Glycolipids activating iNKT cells, such as α-galactosylceramide (αGalCer), can enhance the immune response against co-delivered cancer antigens and have been applied in the design of self-adjuvanting anti-tumor vaccines. In this context, this work focuses on the chemical synthesis of ganglioside tumor-associated carbohydrate antigens (TACAs), namely GM3 and (Neu5Gc)GM3 antigens, their conjugation to αGalCer, and their formulation into liposomes as an efficient platform for their in vivodelivery. Liposomes containing GM3–αGalCer, (Neu5Gc)GM3–αGalCer, and equimolar amounts of the two conjugates have been fully characterized and their ability to activate iNKT cell has been confirmed ex vivoin mouse and human cell assays. The candidates were tested in in vivoimmunization studies, demonstrating an ability to induce both TH1 and TH2 cytokines leading to the production of all subclasses of IgG antibodies. Notably, the study also demonstrated that serum antibodies raised against the two TACAs, alone and in combination, were cross-reactive. This finding has consequences for future vaccine designs – even if a highly tumor-selective antigen is chosen, the resulting antibody response may be broader than anticipated.