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DKK1 long peptide for myeloma immunotherapy
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Figure 3. Dickkopf-1 (DKK1) -long peptide (LP) efficiently induces DKK1-specific CD4+ T-cell response in vivo. Shown are CD4+ T-cell responses induced in HLA-
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LP or DKK1-P30 short peptide. Representative results from one of three independent experiments are shown. (B) Flow cytometry analysis showing the per-
-LP or HLA-DR-restricted P30 short peptide. (A) Carboxyfluorescein succinimidyl ester (CFSE) dilution assay showing the percentages of proliferating CD4+ T cells from the spleen of mice after ex vivo re-stimulation with dendritic cells (DC) pulsed, but not unpulsed, with
DR*4-transgenic mice after immunization with DKK1
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DKK1
centages of IFN-g-expressing or FoxP3+ CD4+ T cells from the spleen of mice after ex vivo re-stimulation with DC pulsed, but not unpulsed, with DKK1 P30 short peptide. Representative results from one of four experiments are shown.
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LP or DKK1-
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C
Figure 4. Dickkopf-1 (DKK1) -long peptide (LP) induces DKK1-specific 3-76
humoral immune response in vivo. Serum titers of DKK1-specific anti-
-LP or short peptide are shown. (A) ELISA showing the titers of DKK1-specific IgG antibodies in
bodies in mice after immunization with DKK1
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LP or P20. (B) ELISA showing the titers of DKK1-specific IgG antibodies in HLA-DR*4-
HLA-A*0201-transgenic mice immunized with DKK1
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LP or P30. (C) Effect of DKK1 on differentiation of human osteoblasts in culture with the addition of con-
transgenic mice immunized with DKK1
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trol mouse IgG, anti-DKK1 antibody (R&D Systems) or serum from DKK1
3- LP vaccinated (after the 4th vaccination) HLA-A*0201-transgenic mice. Results of three experiments are shown. DMSO: dimethyl sulfoxide.
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*P<0.05; **P<0.01.
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