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. 2021 Jul 28;9(7):e001910. doi: 10.1136/jitc-2020-001910

Figure 2.

Figure 2

Hcit-specific immune responses are CD4 mediated and mediate efficient tumor therapy in vivo. IFNγ ELISpot responses were assessed for HHDII/DP4 mice immunized with homocitrullinated peptides. Splenocytes from Aldo74-93Hcit, Aldo140-157Hcit Cyk371-388Hcit, Bip328-346Hcit, Bip562-579Hcit or Eno156-176Hcit immunized mice were restimulated with peptides alone or in combination with anti-CD4 or anti-CD8 blocking antibodies ex vivo (A) or after 7-day culture (B). Data are collated from at least two independent studies where n=3. In vivo tumor survival studies were carried out by implanting mice with HLA-matched B16F1 cells with constitutive MHC-II (B16 HHDII/DP4) or IFNγ-inducible MHC-II (B16 HHDII/iDP4). Mice were then given three immunizations of homocitrullinated or WT peptides. Survival was assessed for aldolase peptides in mice implanted with the B16 HHDII/DP4 (C) and B16 HHDII/iDP4 (D) melanoma lines. In the B16 HHDI/iDP4 model, survival was also determined for Bip328-346Hcit, Bip562-579Hcit and Eno156-176Hcit (E) and Cyk371-388Hcit (F) peptides. Statistical analysis compared survival in immunized mice with unimmunized mice. A representative data set is shown in which for each group n=10 and significant p values are shown. ELISpot, enzyme-linked immunosorbent spot assay; Hcit, homocitrulline; HLA, human leukocyte antigen; IFNγ, interferon γ; MHC-II, major histocompatibility complex class II; WT, wild type.