Single-cell RNA sequencing of antigen specific CD4+ T cells induced by high vs low doses of MHC-II restricted neoantigens

Here, we report the impact of the dose of the MHC-II restricted neoantigens on the phenotypic and the effector function of tumor-specific CD4 T cells. Whereas vaccines containing low doses of MHC-II-restricted neoantigen (LDVax) generated helper CD4 T cells that promote the proliferation and the cytotoxic functions of CD8 T cells, vaccines containing high doses of MHC-II restricted neoantigen (HDVax) elicited type 1 regulatory T cells that suppress the antitumor immunity. T3 tumor-bearing 129S6 male mice were vaccinated on day 6 post-tumor implantation, and 7 days later, tumors were harvested and processed into single-cell suspension. CD4+ T cells were then enriched using positive selection beads and then FACS sorted to ensure high purity (>95%).