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Blood, Vol. 106, Issue 13, 4225-4233, December 15, 2005
Immunomodulatory dendritic cells require autologous serum to circumvent nonspecific immunosuppressive activity in vivo
Blood Haase et al.
106: 4225
Supplemental materials for: Haase et al
Files in this Data Supplement:
- Figure S1: Cytokine production by splenocytes after immunization of naïve mice with mature DCs
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DCs were activated as indicated and BALB/c mice were treated with 106 syngeneic DCs on day -14 and -7. A: Splenocytes were harvested on day 0 and 106 cells were cultured in medium containing 10% FBS for 72 hrs and cytokine production was analyzed. Each dot represents an individual mouse. Shown is also IL-12 p70 production by DCs. B: DCs were analyzed by flow cytometry for CD40 and CD86. Cells are gated on live CD11c+ cells.
- Figure S2: Effect of DC treatment of LCMV-specific immunity in vivo
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Naïve BALB/c mice were injected i.p. with 106 of the indicated DC subset on day -10 and -3, followed by LCMV-infection (105 pfu) on day 0. The LCMV-specific T-cell response was analyzed by intracellular cytokine staining on day 8 after infection. A: Total number of splenocytes. B: Percent CD8+ splenocytes. C: Splenocytes were restimulated in vitro with 10 g/ml NP-118 peptide for 4½ hrs in the presence of brefeldin A and analyzed by ICCS for IFN- as described. *: p<0.05; **: p<0.01 and ***: p<0.005 compared to PBS-group.
- Figure S3: NMS-IL10 DCs induce activation and subsequent deletion of CD8+ T cells
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CFSE-labeled mesenteric LN cells from OT-1 mice (5 × 105) were stimulated with antigen-pulsed DCs (5 × 103) as indicated. A: T cell activation was analysed by CFSE-dilution and stained for Annexin V or OVA-Kb tetramers. B: The number of apoptotic cells in percentage of total dividing cells (i.e. CFSElo cells) is depicted. C: From the total number of cells present in the wells, the total number of OVA-specific CD8+ T cells was calculated. One representative experiment of two is shown.
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