Journal: Cell Reports

Article Title: Notch Signaling Facilitates In Vitro Generation of Cross-Presenting Classical Dendritic Cells

doi: 10.1016/j.celrep.2018.05.068

Figure Lengend Snippet: Notch-Driven Differentiation Yields Functional Human cDC1s DCs generated from CD34 + progenitors in cultures with OP9 or OP9-DL1 were analyzed in parallel to primary DCs from PB. (A and B) Cytokine production by DCs stimulated for 14 hr with a cocktail of TLR agonists (poly-I:C, LPS, CL075, and CpG). (A) Representative flow cytometric analysis of intracellular cytokine production (TNF, IL-12, and IFN-α) in the indicated gated DC subsets from OP9-DL1 cultures. Grey contours represent unstimulated cells; numbers represent the cytokine-positive fraction. (B) Proportion of cytokine-positive cDC1s (purple), cDC2s (red), or pDCs (blue) generated from CD34 + progenitors in culture with OP9 (n = 4) or OP9-DL1 (n = 3) cells compared with PB primary (n = 7) cells following TLR stimulation. Circles, histograms, and bars represent individual experiments, mean, and SEM, respectively; p values are indicated. (C and D) T cell stimulation by DCs cultured with sorted allogeneic blood CD3 + T cells. (C) Representative flow cytometric analysis of T cell proliferation in response to culture with DCs. The data show output of a T cell and cDC1 (generated with OP9-DL1) culture. CD11c + CD141 + cDC1s could be identified (purple gate) and gated out. CD3 + T cells were subdivided by CD8 and CD4 expression. Cell division was indicated by CFSE dilution (turquoise gate). (D) Proportion of CD4 + or CD8 + T cells that underwent division (CFSE dilution) in culture with cDC1 (purple) or cDC2 (red) isolated from PB or generated in culture with OP9 or OP9-DL1 (DL1) cells. T cells cultured alone or with beads coated with anti-CD3 plus anti-CD28 were used as negative (Neg) and positive (Pos) controls, respectively. Responses to blood DCs were generated from 2–3 DC donors and 3 T cell donors (2–6 independent experiments). Responses to cultured DCs were generated from 2 BM donors combined with 3 T cell donors (4–6 independent experiments). Each circle represents an independent experiment (mean of 1–3 technical replicates). Histograms and bars represent mean and SEM, respectively. The p values were derived from unpaired two-tailed Student’s t test.

Article Snippet: Mouse anti-human CD141 (BV510/APC conjugate, clone 1A4/AD5-14H12, 3/5 μl/50 μl sample) , BD/Miltenyi , Cat# 563298, 130-090-907.

Techniques: Functional Assay, Generated, Cell Stimulation, Cell Culture, Expressing, Isolation, Derivative Assay, Two Tailed Test

Journal: Cell Reports

Article Title: Notch Signaling Facilitates In Vitro Generation of Cross-Presenting Classical Dendritic Cells

doi: 10.1016/j.celrep.2018.05.068

Figure Lengend Snippet:

Article Snippet: Mouse anti-human CD141 (BV510/APC conjugate, clone 1A4/AD5-14H12, 3/5 μl/50 μl sample) , BD/Miltenyi , Cat# 563298, 130-090-907.

Techniques: Recombinant, Isolation, Cell Culture, Microarray, Software

Journal: Immunity

Article Title: CD8 + T cell activation in cancer comprises an initial activation phase in lymph nodes followed by effector differentiation within the tumor

doi: 10.1016/j.immuni.2022.12.002

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: anti-human CD141 (FITC) , Miltenyi Biotec , clone: REA674, RRID: AB_2751167.

Techniques: Blocking Assay, Recombinant, Staining, Selection, Enzyme-linked Immunosorbent Assay, Software