Fig. 5: Necroptotic dying cell immunization results in CD4⁺ T Cell priming upon tumor challenge.

A Mice were immunized with NEC and boosted with a second immunization two days later (2× NEC). Mice subsequently received OT-II cells five days later followed by challenge with live tumor cells the next day. At day 9 post-tumor implant the tumor and draining inguinal lymph nodes from the tumor (tdLN) and immunization site (immLN) were collected for flow cytometry. Image was created with BioRender.com. B Total count and C % of CD44^Hi for OT-II CD4⁺ T cells found in the lymph nodes as indicated. Treatment groups were compared using two-way ANOVA. D Total count per gram of tumor, E % of CD44^Hi and (F) % of PD-1^Hi for tumor OT-II CD4⁺ T cells. Treatment groups were compared using unpaired Student’s t-test. For A–F, data is representative of two independent experiments (n = 3–4 per treatment group per experiment). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.