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Blood, Vol. 113, Issue 26, 6593-6602, June 25, 2009
NK cell–mediated killing of target cells triggers robust antigen-specific T cell–mediated and humoral responses
Blood Krebs et al.
113: 6593
Supplemental materials for: Krebs et al
Files in this Data Supplement:
- Table S1. Tissue distribution of γ-irradiated or Kb-deficient splenocytes after injection in C57BL/6J recipient mice (PDF, 48.4 KB)
- Figure S1 (JPG, 288 KB)
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(A) Percentage of IFNγ+ cells within the CD8+ T-cell population after OVA257–264-restimulation of splenocytes isolated from mice immunized with 106 γ-irradiated or live act-mOVA.Kb-deficient cells. (B) Depletion of NK cells by anti–asialo-GM1or anti-NK1.1 (PK136) treatment. Mice were treated at day 1 and 3 and splenocytes were assessed for the presence/absence of NK cells at day 5 using flow cytometry and staining for NKp46, DX5, and NK1.1. (C) Reduced CD8+ T-cell responses in PK136 treated mice 8 days after immunization with 106 live act-mOVA.Kb–deficient cells. (D) CD8+ T-cell responses, as measured by IFNγ production after OVA257–264-restimulation, in mice immunized with various doses of γ-irradiated act-mOVA.Kb+∕+ cells.
- Figure S2 (JPG, 127 KB)
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(A) Killing of class I MHC-deficient target cells in C57BL/6J wildtype control, C57BL/6J-Prf1tm1Sdz, and C57Bl/6Jgdl/gdl recipient mice 48 hours after injection i.v.. (B) Percentage of IFNγ+ cells within the CD8+ T-cell population in spleen after OVA257–264-restimulation in C57BL/6J wildtype, FasL- or perforin-deficient mice previously immunized with 106 γ-irradiated act-mOVA.Kb+∕+ cells.
- Figure S3 (JPG, 208 KB)
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(A) Killing and removal of class I MHC-deficient target cells in wildtype and MyD88/Trif double-deficient mice as measured by flow cytometry, 48 hours after injection of target cells i.v.. (B–E) CD8+ T-cell responses in wildtype, IL-1R–, IL18R-, MyD88-, Trif-, and TLR4-deficient mice. Mice were immunized with 106 live act-mOVA.Kb−∕− splenocytes i.v. and CD8+ T-cell responses were measured at day 8 by intracellular IFNγ production upon restimulation of splenocytes with SIINFEKL peptide ex vivo. (F, G) Class II MHC (F) and CD86 (G) expression on Flt3L-treated bone-marrow derived DCs from wildtype and MyD88/Trif double-deficient mice. DCs were left untreated or activated with LPS or Poly I:C for 24 hours and expression of class II MHC and CD86 was assessed by flow cytometry.
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