dct 180–188 peptide  (Pepscan Inc)

Journal: Scientific Reports

Article Title: Enhancing Immune Responses to Cancer Vaccines Using Multi-Site Injections

doi: 10.1038/s41598-017-08665-9

Figure Lengend Snippet: Survival correlated with the magnitude of antigen-specific blood-derived CD8 + T-cell responses in prophylactic and therapeutic models of metastatic brain cancer. Female C57BL/6 mice had 1,000 B16-F10 melanoma cells stereotactically implanted into the parenchyma of the right hemisphere of the brain. ( A ) Eleven days prior to, or ( B ) five days after intracranial challenge, mice were vaccinated intramuscularly with 1 × 10 8 pfu of recombinant human serotype 5 adenovirus expressing the melanoma-associated antigen dopachrome tautomerase, with injections administered into the semitendinosus of both hind limbs. Blood-derived CD8 + T cells specific for the immunodominant self-epitope DCT 180–188 were quantified fourteen days post-vaccination by flow cytometric assessment of intracellular cytokine staining after ex vivo re-stimulation with peptides (supplementary Figure ). Pearson correlation analysis was performed for overall survival versus frequency (left panel; n = 33 for [A] and 52 for [B]) and total number (right panel; n = 23 for [A] and 52 for [B]; pooled from five and ten experimental replicates, respectively) of antigen-specific CD8 + T cells. Lines of best fit with 95% confidence intervals are shown.

Article Snippet: For quantification of DCT-specific CD8 + T cell responses, cells were re-stimulated ex vivo with the immunodominant peptide that binds to H-2K b (DCT 180–188 ; SVYDFFVWL, which is conserved in human and murine DCT; PepScan Systems, Lelystad, Netherlands).

Techniques: Derivative Assay, Recombinant, Expressing, Staining, Ex Vivo

Journal: Scientific Reports

Article Title: Enhancing Immune Responses to Cancer Vaccines Using Multi-Site Injections

doi: 10.1038/s41598-017-08665-9

Figure Lengend Snippet: Multi-site vaccination increased the magnitude of transgene-specific CD8 + T cells in spleens and lymph nodes. Tumor-free female C57BL/6 mice were vaccinated intramuscularly with 1 × 10 8 pfu of recombinant human serotype 5 adenovirus expressing the melanoma-associated antigen dopachrome tautomerase, with injections spread across one (semitendinosus; left hind limb), two (semitendinosus of both hind limbs) or four (semitendinosus of both hind limbs and the triceps brachii of both forelimbs) sites. ( A , B ) Spleen and ( C , D ) lymph node-derived CD8 + T cells specific for the immunodominant self-epitope DCT 180–188 were quantified fourteen days post-vaccination by flow cytometric assessment of intracellular cytokine staining after ex vivo re-stimulation with peptides. ( A , B ) One-way analysis of variance with Tukey’s multiple comparison test (upper panels) and Pearson correlation analysis (lower panels) was performed for the ( A ) frequency and ( B ) total number of antigen-specific splenic CD8 + T cells ( n = 4/treatment). ( C–D ) Two-way analysis of variance with Tukey’s multiple comparison test (upper panels) and Pearson correlation analysis (lower panels) was performed for the ( C ) frequency and ( D ) total number of antigen-specific lymph node-derived CD8 + T cells ( n = 4/treatment; L = left node, R = right node).

Article Snippet: For quantification of DCT-specific CD8 + T cell responses, cells were re-stimulated ex vivo with the immunodominant peptide that binds to H-2K b (DCT 180–188 ; SVYDFFVWL, which is conserved in human and murine DCT; PepScan Systems, Lelystad, Netherlands).

Techniques: Recombinant, Expressing, Derivative Assay, Staining, Ex Vivo, Comparison