Development of an integrated in vitro and in vivo OVA-specific system for cancer vaccine discovery

We have established an advanced and integrated in vitro and in vivo platform for various types of cancer vaccines, including peptide vaccines, mRNA vaccines, DC vaccines and more. Ovalbumin (OVA) is a key model antigen which has been widely used in vaccination experiments. Here we established an OVA overexpressed system based on different types of tumor cells, including B16F10-OVA and MC38-OVA.  OVA expression was initially confirmed via Western Blot analysis. Then, we showed that MHC-I:OVA complexes were presented on the surface of stable cell lines through IFN stimulation. The results of in vitro CD8+ T cell activation and killing assays showed that the T cells can be activated and target cells were specifically killed after incubation, which indicates that the presented OVA peptides can be utilized as an effective therapeutic target.

OVA RNA vaccine was prepared by mixing OVA mRNA with special transfection reagents. Using an ELISPOT assay, we demonstrated the immunogenicity of an OVA-specific mRNA vaccine. Furthermore, an in vivo tumor model of B16F10-OVA was established and the anti-tumor effect of the vaccine was evaluated. The mechanism of tumor growth inhibition was also investigated via T cell, B cell and other cell population depletions. Additionally, we explored the target protein biodistribution and in vivo profile of mRNA vaccine carrier with a Lumina imaging system. In summary, this integrated OVA-specific system can be used for new adjuvants and peptide/mRNA carrier exploration in cancer vaccine discovery.

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