Journal: eLife

Article Title: A tissue-like platform for studying engineered quiescent human T-cells’ interactions with dendritic cells

doi: 10.7554/eLife.48221

Figure Lengend Snippet: ( a ) Co-culture of 1G4 -expressing naïve CD8 T-cells (magenta), 6F9 -expressing naïve CD4 T-cells (cyan) and acDCs loaded with 100 nM NY-ESO-6T and 10 μM MAGE-A3 243-258 peptides (green), , in collagen gels containing CXCL12. ( b ) Upregulation of CD69 by CD4 (top) and CD8 T-cells (bottom), and the upregulation of the costimulatory molecule CD86 on acDCs (middle) in the co-culture system from ( a ) in the presence and absence of MAGE as a stimulus for CD4 T-cells. Representative of >3 independent repeats. In all experiments, the cell ratio was 5:5:1 (CD4: CD8: acDC). ( c ) 1G4 -expressing memory CD8 T-cells (magenta) co-cultured with acDCs loaded with 100 nM NY-ESO-9V (green). Cells form clusters around acDC in the presence of anti PD-1 blocking antibody Nivolumab (left) compared to less tight contacts in the isotype control (right). ( d ) The activation of the CD8 T-cells by upregulation of CD69, 41BB as well as cytotoxicity measured as the release of LDH following the killing of the target. In all experiments, the cell ratio was 10:1 (CD8: acDC). Scale bars = 10 μm.

Article Snippet: The following anti-human antibodies were purchased from BD Bioscience: CD45RA (HI100), CD4 (RPA-T4), and CD11c (B-ly6), CD14 (MΦP9) and PD-1 (EH12.2H7).

Techniques: Co-Culture Assay, Expressing, Cell Culture, Blocking Assay, Activation Assay

Journal: STAR Protocols

Article Title: Simplified mass cytometry protocol for in-plate staining, barcoding, and cryopreservation of human PBMC samples in clinical trials

doi: 10.1016/j.xpro.2022.101362

Figure Lengend Snippet: Surface antibody master mix, titrated for staining of up to 3 M cells per condition

Article Snippet: Anti-Human CD279/PD-1 (EH12.2H7)-174Yb—100 Tests (1:100) , Fluidigm , Cat#3174020B.

Techniques: Staining

Journal: STAR Protocols

Article Title: Simplified mass cytometry protocol for in-plate staining, barcoding, and cryopreservation of human PBMC samples in clinical trials

doi: 10.1016/j.xpro.2022.101362

Figure Lengend Snippet:

Article Snippet: Anti-Human CD279/PD-1 (EH12.2H7)-174Yb—100 Tests (1:100) , Fluidigm , Cat#3174020B.

Techniques: Recombinant, Saline, Staining, Blocking Assay, Isolation, Software, Cell Counting, Cytometry

Journal: Cell Reports Medicine

Article Title: OX40 agonism enhances PD-L1 checkpoint blockade by shifting the cytotoxic T cell differentiation spectrum

doi: 10.1016/j.xcrm.2023.100939

Figure Lengend Snippet:

Article Snippet: Anti-human PD-1 (clone EH12.2H7) (175Lu) , Fluidigm , Cat# 3175008; RRID: AB_2687629.

Techniques: Recombinant, Purification, Staining, Cell Isolation, Transgenic Assay, Software, Gene Expression