Figure 1.

Therapeutic vaccination with variable epitope libraries (VELs) inhibits 4T1 tumor growth and the development of spontaneous metastases. (a) Groups of 4T1 tumor‐bearing BALB/c mice were immunized (at day 5) intravenously (i.v.) with 2·5 × 10¹² recombinant M13 phage particles, expressing four vaccines bearing combinatorial VELs, based on full‐length (VEL‐Full SVN), N‐terminal (VEL‐NT), core (VEL‐Core) and C‐terminal (VEL‐CT) sequences of surviving (SVN), respectively (figure created with BioRender.com). (b–e) Tumor growth was monitored by measuring tumor volumes. (f–i) Lung macrometastases were counted at day 33. Mice were also vaccinated with the corresponding wild‐type (WT) counterparts of the four VELs. Tumor‐bearing untreated mice and mice vaccinated with an unrelated phage (U.E.) were used as controls. (j–m) Lungs from each group were fixed in Bouin’s solution, representative images are shown. (b–e) Tumor volumes are presented as mean ± 95% CI mm³; n = 30, *P < 0·033, **P < 0·02, ***P < 0·001. Two‐way analysis of variance for repeated measurements and Tukey post‐hoc test for multiple comparisons were used. (f–i) Lung macrometastases are presented as mean ± 95 CI; n = 30; *P < 0·033, **P < 0·02, ***P < 0·001. One‐way analysis of variance with Tukey post‐hoc test for multiple comparisons was used.