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. 2009 Jun 3;4(6):e5739. doi: 10.1371/journal.pone.0005739

Figure 3. C57BL/6 mice are tolerized to HuD.

Figure 3

(a) C57BL/6 mice were immunized with AdVHuD or AdVβgal+PTx (2 mice per group). 13 days later, CD8+ T cells were isolated from the spleen and plated in an IFNγ ELISPOT assay (2×105/well) with EL4 pulsed with 10 uM peptide (5×104/well). The assay was performed in triplicate. Means are plotted and error bars represent standard deviations of the mean. Data is representative of four experiments. (b) C57BL/6 mice were immunized with AdVHuD−/+PTx (2 mice per group). 13 days later, splenocytes were stimulated in vitro with 0.5 uM HuD p321. On day 7, CD8+ T cells were plated in an IFNγ ELISPOT assay (104/well) with DC pulsed with 10 uM peptide (7×103/well). The assay was performed in triplicate. Means are plotted and error bars represent standard deviations of the mean Data is representative of two experiments. (c) Individual HuD+/+ or HuD−/− mice were immunized with AdVHuD+PTx and used in an IFNγ ELISPOT assay as described in (a). The assay was performed in triplicate. Means are plotted and error bars represent standard deviations of the mean. Data is representative of four experiments. (d) Half of the spleens from mice immunized in (c) were stimulated in vitro with HuD p321. After 7 days, CD8+ T cells were isolate from stimulation cultures and plated in an IFNγ ELISPOT (104/well) with peptide pulsed EL4 cells (5×104/well).