Journal: iScience

Article Title: PD-L1 + CD8 + T cells enrichment in lung cancer exerted regulatory function and tumor-promoting tolerance

doi: 10.1016/j.isci.2022.103785

Figure Lengend Snippet: The IMC system analysis indicated PD-L1 + CD8 + T cells enrichment in lung tumor tissues and spatially located with PD-1 + effect T cells (A) Staining for tumor cells (Pan-CK), immune cells (CD45, CD3, CD4, and CD8), and nuclei (DNA) in one of the representative patients. (B) Normalized expression of CD45, Pan-CK, Epcam, CD3, CD4, and CD8 was shown in the viSNE plot. (C) The intensity of Pan-CK + , Epcam + , CD45 + , and CD3 + cells in lung tumors and adjacent tissues, N = 9. (D) Representative co-expression of CD8 and PD-L1 in tumor tissues from two patients. (E) viSNE plot of CD8 + T cells, showing the identification of 11 clusters. Each dot corresponds to a single cell, colored according to the cell cluster. (F) Heatmap of the Phenograph clusters of CD8 + cells. The relative expression levels of markers across cells were shown. (G) The frequency of PD-L1 + , PD-1 + , CD38 + , and CD39 + in CD8 + T cells in adjacent and tumor tissues is determined by IMC. N = 9, two tailed paired t-test. (H) Two representative samples of the neighborhood cells with PD-L1 + CD8 + T cells in lung tumor tissues. (I) viSNE plot showed the neighborhood cells with PD-L1 + CD8 + T cells in lung tumor tissues, including 7 clusters. (J) viSNE plot of the related markers of the CD8 + T cell clusters generated in (I). (K) Re-cluster of neighborhood CD8 + T cells with PD-L1 + CD8 + T cells in lung tumor tissues, including five clusters. (L) Heatmap of the related markers across the neighborhood CD8 + T cells generated in (K).

Article Snippet: anti-human CD279 (PD-1) -155Gd , Fluidigm , Cat# 3155009B.

Techniques: Staining, Expressing, Two Tailed Test, Generated

Journal: iScience

Article Title: PD-L1 + CD8 + T cells enrichment in lung cancer exerted regulatory function and tumor-promoting tolerance

doi: 10.1016/j.isci.2022.103785

Figure Lengend Snippet: PD-L1 + CD8 + T cells engage in regulatory activity in vitro (A, B, C, and D) CD8 + T cells were cocultured with lung cancer cell line HCC827 or alone for 48 h. (A) FACS analysis of PD-L1, CD38, and PD-1 expression. One of three similar experiments is shown. (B) PD-L1 and CD38 expressions were evaluated by FACS. One of three similar experiments is shown. TNFα (C) and IFNγ (D) expression was compared between PD-L1 + CD38 lo and PD-L1 - CD38 hi CD8 T cells. One of three similar experiments was shown. (E) The PD-1 expression of PD-L1 + CD38 lo and PD-L1 - CD38 hi CD8 T cells was shown. One of five similar experiments is shown. (F, G, and H) CD8 + T cells were cocultured with the lung cancer cell line HCC827 for 48 h, and PD-L1 + and PD-L1- CD8 + T cells were sorted and then cocultured with CFSE-labeled CD8 + T cells for 96 h. One of three similar experiments is shown. (F) The proliferation (G) TNFα and IFNγ expression of CFSE-labeled CD8 + T cells were measured. One of three similar experiments was shown. (H) CD8 + T cells were cocultured with the lung cancer cell line HCC827 for 48 h, and PD-L1+ and PD-L1- CD8 + T cells were sorted and then cocultured with CFSE-labeled CD8 + T cells for 72 h in the presence of an anti-PD-L1 antibody or IgG control. TNFα and IFNγ expression was measured. One of three similar experiments was shown. All the data were presented as the mean ± SEM, and the p value was calculated by a two-tailed Student’s t-test, ∗p <0.05, ∗∗p <0.01.

Article Snippet: anti-human CD279 (PD-1) -155Gd , Fluidigm , Cat# 3155009B.

Techniques: Activity Assay, In Vitro, Expressing, Labeling, Two Tailed Test

Journal: iScience

Article Title: PD-L1 + CD8 + T cells enrichment in lung cancer exerted regulatory function and tumor-promoting tolerance

doi: 10.1016/j.isci.2022.103785

Figure Lengend Snippet:

Article Snippet: anti-human CD279 (PD-1) -155Gd , Fluidigm , Cat# 3155009B.

Techniques: Recombinant, Antibody Labeling, Enzyme-linked Immunosorbent Assay, Sequencing, Software