easyseptm mouse cd8+ t cell enrichment kit  (STEMCELL Technologies Inc)

Journal: Cancer Immunology Research

Article Title: High Levels of Endogenous Omega-3 Fatty Acids Promote Dendritic Cell Antigen Presentation and Improve Dendritic Cell–Based Cancer Vaccine Efficacy in Mice

doi: 10.1158/2326-6066.CIR-24-0927

Figure Lengend Snippet: FAT-1 transgenic mice show improved overall survival, lower body mass, and protection against tumor growth compared with WT mice. A, Schematic of the FAT-1 transgenic mouse model. B, Kaplan–Meier curve showing the survival probability of WT and FAT-1 naïve animals. C, Comparison of body weight of WT and FAT-1 animals as they age (****, P < 0.0005). D, Tumor growth kinetics of TC-1 lung cancer cell lines in FAT-1 and WT (*, P < 0.02). E, Kaplan–Meier survival curve for TC-1 lung cancer tumor–bearing WT and FAT-1 mice (**, P < 0.009). F, Tumor growth kinetics of B16F10 melanoma cancer cell lines in WT and FAT-1 animals (***, P < 0.0001). G, Kaplan–Meier survival curve for B16F10 melanoma tumor–bearing WT and FAT-1 mice (*, P < 0.002). H, Percentage of tumor-free mice after inoculation in WT vs. FAT-1 in the B16F10 melanoma tumor model (****, P < 0.005). I, Illustration showing the experimental design of BMDC culture and antigen presentation assay in vitro . J, The bar graph showing the frequency of major immune cell types in the spleen of WT vs. FAT-1 mice (orange, stained control; green, FAT-1; blue, WT). K, Levels of IFNγ in supernatant from WT or FAT-1 BMDCs cocultured with CD8 + T cells from PMEL-1 mice measured by ELISA (*, P < 0.008). L, Overlay of expression of major surface markers, including activation and costimulatory markers (CD11b, CD11c, MHC class II, CD40, CD80, CD86) between mature antigen-pulsed BMDCs derived from FAT-1 (green curve) and WT (blue curve) mice. Statistical analysis: Multiple comparisons were made using two-way ANOVA. The Šidák correction test was applied to correct for multiple comparisons. The Mann–Whitney test (unpaired/nonparametric comparing the difference in median) was used as applicable for all figures. EPA, eicosapentaenoic acid; PMA/ION, phorbol 12-myristate 13-acetate/ionomycin.

Article Snippet: The CD8 + T cells were isolated by magnetic bead isolation using the Miltenyi mouse CD8a T-cell enrichment kit according to the manufacturer’s protocol.

Techniques: Transgenic Assay, Comparison, Immunopeptidomics, In Vitro, Staining, Control, Enzyme-linked Immunosorbent Assay, Expressing, Activation Assay, Derivative Assay, MANN-WHITNEY

Journal: Cancer Immunology Research

Article Title: High Levels of Endogenous Omega-3 Fatty Acids Promote Dendritic Cell Antigen Presentation and Improve Dendritic Cell–Based Cancer Vaccine Efficacy in Mice

doi: 10.1158/2326-6066.CIR-24-0927

Figure Lengend Snippet: FAT-1–derived BMDCs induce a higher vaccine-specific response ex vivo . A, Illustration showing the layout of peptide restimulation of whole blood ex vivo . B and C, Bar graph comparing the frequency of IFNγ + CD8 T cells ( B ) and E7 tetramer + IFNγ + CD8 T cells ( C ) in the overnight restimulation of whole blood with E7 peptide vs. vehicle control across different vaccination groups. D, Bar graph comparing the frequency of PD-1 + CD8 T cells in 48 hours of restimulation of whole blood with E7 peptide vs. vehicle control across different vaccination groups. E–G, Simple regression analysis showing the correlation of the frequencies of IFNγ + CD8 T cells ( E ), E7 tetramer + IFNγ + CD8 T cells ( F ), and PD-1 + CD8 + T cells ( G ) in peptide restimulation with tumor volumes in different vaccination groups. *, P < 0.05; **, P < 0.005; ****, P < 0.0005; NS, nonsignificant. veh, vehicle.

Article Snippet: The CD8 + T cells were isolated by magnetic bead isolation using the Miltenyi mouse CD8a T-cell enrichment kit according to the manufacturer’s protocol.

Techniques: Derivative Assay, Ex Vivo, Control

Journal: Cancer Immunology Research

Article Title: High Levels of Endogenous Omega-3 Fatty Acids Promote Dendritic Cell Antigen Presentation and Improve Dendritic Cell–Based Cancer Vaccine Efficacy in Mice

doi: 10.1158/2326-6066.CIR-24-0927

Figure Lengend Snippet: FAT-1 BMDCs have higher T-cell dwell time, and supplementation of omega-3 in WT BMDCs mimics FAT-1–like antigen presentation ability in vitro . A, Dot blot showing the differences between the expression intensity for individual cytokines/chemokines (duplicate dots for each) between immature BMDCs from FAT-1 and WT. B, Cytokine/chemokine profile in supernatant taken from immature BMDCs. C, Cytokine/chemokine profile of serum from WT or FAT-1 naïve mice. D, Image showing the FAT-1 and WT BMDC coculture with PMEL-1 CD8 T cells stained with viability dye (green, T cells stained with CMFDA, and red, DCs stained with CMPTX). E, Time spent by an antigen-specific T cell with an antigen-loaded DC in an overnight coculture assay. F, Track length generated by T cells during coculture assay (****, P < 0.0005). G, Schematics of BMDC culture in the presence of DHA and resolvins. H–K, IFNγ production by the LPS/hgp100 antigen–pulsed BMDCs grown in the presence of DHA, resolvin D1 (RvD1), RvE1, and RvE4, respectively, when cocultured with antigen-specific CD8 + T cells, measured by ELISA. **, P < 0.005; ****, P < 0.0005; NS, nonsignificant. PMA/ION, phorbol 12-myristate 13-acetate/ionomycin.

Article Snippet: The CD8 + T cells were isolated by magnetic bead isolation using the Miltenyi mouse CD8a T-cell enrichment kit according to the manufacturer’s protocol.

Techniques: Immunopeptidomics, In Vitro, Dot Blot, Expressing, Staining, Co-culture Assay, Generated, Enzyme-linked Immunosorbent Assay