Supplementary Materials

Supplementary Material for:

In Vivo–Generated Antigen-Specific Regulatory T Cells Treat Autoimmunity Without Compromising Antibacterial Immune Response

Shimpei Kasagi, Pin Zhang, Li Che, Brittany Abbatiello, Takashi Maruyama, Hiroko Nakatsukasa, Peter Zanvit, Wenwen Jin, Joanne E. Konkel, WanJun Chen*

*Corresponding author. E-mail: wchen@mail.nih.gov

Published 18 June 2014, Sci. Transl. Med. 6, 241ra78 (2014)
DOI: 10.1126/scitranslmed.3008895

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Schematic design of the study. Therapy of EAE and type diabetes by generation of auto-antigen-specific Foxp3+ Treg cells in vivo.
  • Fig. S2. γ-Irradiation induces immune cell apoptosis in vitro and in vivo.
  • Fig. S3. Gating control of CD4+ lymphocytes stained with isotype control antibodies of IL-17 and IFN-γ.
  • Fig. S4. CD4+Foxp3+ Treg cells increased in the spleen of pPLP-immunized SJL mice treated with irradiation plus phagocytes and pPLP (IRR + MΦ + PLP).
  • Fig. S5. The function of professional phagocytes in apoptosis-antigen–mediated prevention of remitting-relapsing EAE in SJL mice.
  • Fig. S6. In vivo treatment with anti-CD20 and anti-CD8a antibodies depleted B and CD8+ T cells in vivo.
  • Fig. S7. In vivo treatment with anti-CD20 and anti-CD8α antibodies did not affect the frequency of CD4+ T cells in the whole lymphocytes.
  • Fig. S8. Combination of B cell and CD8+ T cell depletion and peptide administration prevented EAE in SJL mice.
  • Fig. S9. NOD mice treated with irradiation plus phagocytes and GAD65 peptide (IRR + MΦ + GAD65) showed decreased GAD65-specific T cell response compared to untreated (PBS) or GAD65-treated (GAD65) mice.
  • Fig. S10. TH17 cells were undetectable in NOD mice.
  • Fig. S11. The total number of T cells in the spleen was comparable between tolerized mice and untreated mice.
  • Fig. S12. TGFβ plays a key role in apoptosis-antigen combined therapy of EAE.
  • Fig. S13. MOG38–49 tetramer+ Foxp3+ Treg cells increased in the spinal cords of apoptosis-antigen–treated EAE mice.
  • Fig. S14. MT-driven T cell proliferation and IFN-γ and IL-17 production in IRR + MΦ + MOG + contrl Ab–treated mice showed similar levels as those in untreated mice.
  • Fig. S15. Cell membrane–bound TGFβ1 of macrophages and Treg cells were increased in EAE mice treated with IRR + MΦ + MOG.
  • Fig. S16. TGFβ is required for the generation of antigen-specific Treg cells in established EAE.
  • Fig. S17. Generation of antigen-specific CD4+CD25+ Treg cells in mice with long-term remission of EAE induced by apoptosis-antigen treatment in the prevention models.
  • Fig. S18. Generation of antigen-specific CD4+CD25+ Treg cells in mice with long-term remission of T1D induced by apoptosis-antigen treatment.
  • Fig. S19. Antigen-specific Treg cells were converted from naïve CD4+ cells in vivo.
  • Fig. S20. Nrp-1 negative MOG38–49 + Foxp3+ T cells were increased in EAE mice after IRR + MΦ + MOG treatment.
  • Fig. S21. Antigen-specific Treg cells were converted from naïve CD4 cells in vivo.

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Other Supplementary Material for this manuscript includes the following:

  • Table S1. Original data values (provided as a separate Excel file).