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. Author manuscript; available in PMC: 2022 Sep 23.
Published in final edited form as: Science. 2021 Aug 27;373(6558):1040–1046. doi: 10.1126/science.abc9113

Fig. 3. SagA improves checkpoint inhibitor immunotherapy and elicits an adaptive immune response.

Fig. 3.

(A) B16-F10 tumor growth in antibiotic-treated mice that were supplemented with the indicated enterococci and treated with anti–PD-L1 starting on day 9. n = 8 mice per group. (B) MCA205 tumor growth in antibiotic-treated mice that were supplemented with the indicated enterococci and treated with anti–PD-1 starting on day 5. n = 8 mice per group. (C) MC38 tumor growth in antibiotic-treated mice that were supplemented with the indicated enterococci and treated with anti–CTLA-4 starting on day 7. n = 8 mice per group. (D) to (I) Quantification of tumor infiltrating CD45+ cells (D), total CD3+ T cells (E), FoxP3+ regulatory T cells (F), CD8+ T cells (G), granzyme B+ CD8+ T cells (H), and OVA-specific CD8+ T cells (I) from B16-OVA tumors in mice supplemented with E. faecalis (Efs) or Efs-sagA harvested five days after the start of anti–PD-L1 treatment by flow cytometry. Data are pooled from two independent experiments of 7-10 mice per group per experiment; each symbol represents one mouse. (J) B16-F10 tumor growth in antibiotic-treated Nod2+/− or Nod2−/− mice that were supplemented with Efs or Efs-sagA and treated with anti–PD-L1 starting on day 7. n = 9-11 mice per group. (K) Western blot detection of ectopically expressed Efm Com15 SagA in secreted protein and cell pellet fractions harvested from overnight cultures of the indicated engineered Lactococcus lactis (Lls) strains using antiserum raised against Efm Com15 SagA. Bottom panels show total protein loading. Numbers indicate estimated molecular weight (kDa). WT = wild-type, ΔSS = signal sequence deletion. (L) B16-F10 tumor growth in antibiotic-treated mice that were supplemented with the indicated Lactococcus lactis (Lls) strains and treated with anti–PD-L1 starting on day 7. n = 9-11 mice per group. Data for (A)-(C) and (K)-(L) represent mean ± s.e.m. and analyzed by mixed-effects model with Tukey’s multiple comparisons post-hoc test. Data for (D)-(I) represent mean ± s.e.m. and analyzed by Mann-Whitney U test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns = not significant.