Science Advances

Supplementary Materials

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  • Methods
  • Fig. S1. Extended phenotyping of Treg population in LNs of normal mice, in TDLNs of mice with B16F10 tumors, and in the disaggregated tumors themselves.
  • Fig. S2. Role of IDO, PD-1, and mTOR pathways during Treg activation in vitro.
  • Fig. S3. In vivo phosphorylation status of S6 and Akt473 in Tregs from TDLNs of tumor-bearing mice, after vaccination in the presence or absence of IDO inhibitor drug (2 mg/ml in drinking water, or vehicle control, as indicated).
  • Fig. S4. Confirmation of specificity of the FoxO3a staining antibody (rabbit monoclonal, clone 75D8, Cell Signaling Technology).
  • Fig. S5. Foxo3a in activated Tregs in vitro.
  • Fig. S6. Activation step cocultures using IDO+ TDLN DCs were supplemented with excess tryptophan at the concentrations shown.
  • Fig. S7. IDO was important in controlling Akt phosphorylation, but it was unclear how IDO, which was expressed in the DCs, influenced the mTOR→Akt473 pathway expressed in Tregs.
  • Fig. S8. Role of PD-1 in maintaining IDO-induced suppressor activity.
  • Fig. S9. Validation of PTENTreg-KO mice.
  • Fig. S10. Growth of EL4-OVA (E.G7) and LLC tumors in PTENTreg-KO hosts.
  • Fig. S11. Spontaneous reprogramming of Tregs in tumors of PTENTreg-KO mice.
  • Fig. S12. Inflammatory intratumoral milieu in E.G7 tumors grown in PTENTreg-KO hosts.
  • Fig. S13. Artifactual preactivation by CD8 bead isolation obscures immunosuppression by tumors.
  • Fig. S14. Tregs were activated using IDO+ TDLN DCs (left) or with αCD3 mitogen and IDO blocked (right).
  • Fig. S15. Effect of PTEN blockade or genetic deletion on response to vaccination.
  • Fig. S16. Absence of autoantibodies in mice receiving VO-OHpic.
  • Fig. S17. Combination of VO-OHpic with indoximod.
  • Fig. S18. Comparison of VO-OHpic versus CAL-101 when given in combination with CTX.
  • Fig. S19. B16F10 tumors were grown in mice bearing an insertion of a fusion protein of GFP with the human DTR into the Foxp3 locus (63).
  • Fig. S20. Requirement for IL-6 receptor expression on Tregs for Treg destabilization and anti-tumor response.
  • Fig. S21. PTENTreg-KO mice lose any further effect of the VO-OHpic PTEN inhibitor.

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