Journal: Cancers

Article Title: Dihydrotestosterone Enhances MICA-Mediated Immune Responses to Epstein-Barr Virus-Associated Gastric Carcinoma.

doi: 10.3390/cancers16183219

Figure Lengend Snippet: Figure 4. DHT increases the cytotoxicity of PBMCs. (A) Fresh PBMCs were treated (stimulated) with EBV EBNA1 full peptide (5 g/mL) and LMP2A full peptide (5 g/mL) for 14 days to establish EBNA1 PBMCs and LMP2A PBMCs, respectively. The proportion of CD8+ T cells in each of these two PBMCs was investigated by FACs analysis. The bright cyan letters and numbers in the dot plot represent the intensity and percentage of antibody binding to the antigen. Specifically, UL4 indicates the percentage of CD8-positive and CD3-negative cells, LL4 represents the percentage of CD8-negative and CD3-negative cells, UR4 corresponds to the percentage of CD8-positive and CD3-positive cells, and LR4 denotes the percentage of CD8-negative and CD3-positive cells. 1e1 means 1 × 101, which equals 10. 1e2 means 1 × 102, which equals 100. 1e3 means 1 × 103, which equals 1000. (B) The proportion of cells secreting INF-γ in each EBNA1 PBMCs and LMP2A PBMCs was examined by FACs analysis. (C) Investigation of cytotoxicity of EBNA1 PBMCs against SNU719 cells treated with 100 nM DHT for 24 h by LDH release-based cytotoxicity assay. Cytotoxicity of EBNA

Article Snippet: The EBV EBNA-1 peptide pool (#130-093- 613, Miltenyi Biotec, Bergisch Gladbach, Germany) was used to generate EBNA-1-specific PBMCs (named EBNA1 PBMCs), while the EBV LMP2A peptide pool (130-093-615, Miltenyi Biotec) was used to generate LMP2A-specific PBMCs (named LMP2A PBMCs).

Techniques: Binding Assay, Cytotoxicity Assay

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: A method for polyclonal antigen-specific T cell-targeted genome editing (TarGET) for adoptive cell transfer applications

doi: 10.1016/j.omtm.2023.06.007

Figure Lengend Snippet: Transduction efficiencies and phenotype differences between expanded cells (A) Transduction efficiencies for pepmix-stimulated vs. anti-CD3/CD28 dynabeads-stimulated T cells for bulk CD3+, CD4+, and CD8+ cells, respectively; n = 6, shown are medians with range. (B) CD4 vs. CD8 proportions within different populations of expanded pepmix-stimulated T cells; n = 6, shown are means with SD. (C) Memory phenotypes and (D) exhaustion marker expression of expanded pepmix-stimulated T cells among wild-type (WT), RNP-only transfected and transduced bulk CD3+, CD4+, and CD8+ cells; n = 4. For (B)–(D), asterisks represent statistically significant differences (∗p < 0.05, ∗∗p < 0.005, two-way ANOVA, Tukey’s multiple comparisons test). ANOVA, analysis of variance; EBV, Epstein-Barr virus; RNP, ribonucleoprotein; AAV, adeno-associated virus; Temra, terminally differentiated; Tem, effector memory; Tcm, central memory; Tscm, stem cell memory T cells.

Article Snippet: PBMCs were stimulated with either anti-CD3/CD28 Dynabeads (Thermo Fisher Scientific) according to the manufacturer’s instructions or with EBV pepmix (PepTivator EBV Consensus peptide pool [Miltenyi Biotec, Bergisch Gladbach, Germany]), at a final concentration of 60 pmol/peptide/mL in CTLm supplemented with 400 U/mL IL-4 and 10 ng/mL IL-7 (R&D Systems, Minneapolis, MN) for 3 days.

Techniques: Transduction, Marker, Expressing, Transfection, Virus

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: A method for polyclonal antigen-specific T cell-targeted genome editing (TarGET) for adoptive cell transfer applications

doi: 10.1016/j.omtm.2023.06.007

Figure Lengend Snippet: Specificity and functionality of pepmix-stimulated and expanded transduced T cells (A) Production of cytotoxicity markers and cytokines (CD107a, Granzyme B, IFNγ, and TNFα) among bulk CD3+, CD4+, and CD8+ populations in response to EBV-pepmix-restimulation, n = 4, means with standard deviation (SD). Asterisks represent statistically significant differences (∗p < 0.05, ∗∗p < 0.005, two-way ANOVA, Tukey’s multiple comparisons test). (B) Six-hour cytotoxicity assay against autologous EBV-transformed LCLs (effector/target = 20:1), means of triplicates with SD for three donors, two-way ANOVA mixed effects analysis, ∗∗ = 0.0043, ∗ = 0.0405, α = 0.05. ANOVA, analysis of variance; EBV, Epstein-Barr virus; WT, wild type; RNP, ribonucleoprotein; AAV, adeno-associated virus; GrB, granzyme B.

Article Snippet: PBMCs were stimulated with either anti-CD3/CD28 Dynabeads (Thermo Fisher Scientific) according to the manufacturer’s instructions or with EBV pepmix (PepTivator EBV Consensus peptide pool [Miltenyi Biotec, Bergisch Gladbach, Germany]), at a final concentration of 60 pmol/peptide/mL in CTLm supplemented with 400 U/mL IL-4 and 10 ng/mL IL-7 (R&D Systems, Minneapolis, MN) for 3 days.

Techniques: Standard Deviation, Cytotoxicity Assay, Transformation Assay, Virus