Fig. 4: Tetravalent anti-mCD27 generates superior vaccine responses compared to its bivalent counterpart.

A–D C57BL/6 mice were inoculated with s.c. B16-OVA melanoma on day 0 and treated with OVA and the indicated antibodies on day 1. Expansion of endogenous antigen-specific CD8⁺ T cells was monitored by obtaining peripheral blood samples from day 0 to day 12 and staining with anti-CD8 and H2-K^b - OVA_257–264 tetramer (tetramer⁺). A Representative flow cytometry contour plots and B compiled data of the antigen-specific T cell response on day 7, with each bar representing the mean ± SEM from 12 individual mice (n = 12, isotype mIgG1, anti-mCD27 mIgG1, tetra anti-mCD27 mIgG1) across 2 independent experiments or 6 individual mice (n = 6, anti-mCD27 mIgG1 NQ, tetra anti-mCD27 mIgG1 NQ) from 1 experiment. Statistical significance was determined using one-way ANOVA, with Tukey’s post-hoc test for multiple comparisons, and significance values are indicated in the figure. C Time course of the antigen-specific T cell response, with each data point representing the mean ± SEM (n = 12, isotype mIgG1, anti-mCD27 mIgG1, tetra anti-mCD27 mIgG1; n = 6, anti-mCD27 mIgG1 NQ, tetra anti-mCD27 mIgG1 NQ). Statistical significance was determined by calculating the AUC and performing one-way ANOVA on AUC values, with significance values indicated in the figure. D Tumor area for mice inoculated with B16-OVA cells on day 0 and treated with OVA and the indicated antibodies on day 1. Data points are the mean ± SEM of 12 individual mice (n = 12, isotype mIgG1, anti-mCD27 mIgG1, tetra anti-mCD27 mIgG1) across 2 independent experiments or 6 individual mice (n = 6, anti-mCD27 mIgG1 NQ, tetra anti-mCD27 mIgG1 NQ) from 1 experiment. Statistical significance by one-way ANOVA was performed on AUC, with significance values indicated in the figure. Responses of “NQ” antibody variants were not statistically significant compared to the isotype control. Source data are provided as a Source data file.