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. 2022 Feb 22;10(2):e003978. doi: 10.1136/jitc-2021-003978

Figure 2.

Figure 2

Immunogenicity of mutated HLA-A*02.01-restricted peptides in HHD-DR1 mice. (A) HHD-DR1 mice (n=3–6/group) were immunized with individual peptides plus poly(I:C) and anti-CD40 adjuvants. Ten days later splenocytes were stimulated with the mutated or WT version of each peptide and responses measured by ELISPOT. (B) Selected immunogenic peptides (n=3 mice/peptide) were also tested against decreasing concentrations of mutated and WT peptides. (C) Splenocytes from the same immunized mice were stimulated with the mutated and WT version of the peptides and expression of IFN-γ, TNF-α and CD107 in CD8 T cells were measured by flow cytometry. Representative example of peptide KYV (left panels) and summarized results (right panels) are shown. (D) HHD-DR1 mice (n=4/group) were immunized with longer versions (containing four extra amino acids at each end) of neoAg peptides and splenocytes were stimulated in vitro with the long peptide and the minimal epitope. (E) Splenocytes from mice immunized with peptide KYV (n=4) were stimulated with the peptide or with HEK-293 cells transfected with a plasmid encoding an elongated version of KYV (pSP/eKYV/MITD) or mock transfected (control). (F) Splenocytes from mice (n=4) immunized with peptide KYV were stimulated with the peptide or with HepG2 cells transfected with a plasmid encoding an elongated version of neoAgs LKR, ALL, QQW, KVY and EVT (pSP/neoAgs/MITD) or mock transfected (control). TNBC, too numerous to be counted. *p< 0.05.