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: 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

Journal: Journal of Inflammation Research

Article Title: Tumor-Infiltrating PD-L1+ Neutrophils Induced by GM-CSF Suppress T Cell Function in Laryngeal Squamous Cell Carcinoma and Predict Unfavorable Prognosis

doi: 10.2147/JIR.S347777

Figure Lengend Snippet: Immune cell infiltration profiles in LSCC. ( A ) Representative polychromatic dot plots displayed CD45+ leukocytes, CD3+ T cells, CD4+ T cells, CD8+ T cells γδT cells (γδTCR+) and neutrophils (CD66b+) in peripheral blood, tumor tissues and normal tissues from LSCC patients or healthy controls. ( B ) Calculative distribution of lymphocytes, neutrophils, NLR, CD3+T cells, CD4+T cells, CD8+T cells, γδT cells, and CD4+T cells /CD8 + T cells ratio in peripheral blood, tumor tissues and normal tissues. ( C ) Representative PD-1 expression on CD4+T/CD8+T/γδT cells and PDL-1 expression on CD66b+ neutrophils from the peripheral blood, tumor tissues and normal tissues. ( D ) Statistics analysis of PD-1+ CD4+T, PD-1+ CD8+T, PD-1+ γδT cells and PDL-1+ CD66b+ neutrophils. ( E ) Association analysis of TANs, NLR, CD3+ T cells or CD8+ T cells and tumor staging. ( F ) Linear correlation between TANs and CD3+ T cells, CD4+T cells, CD8+ T cells, or γδT cells. ( G ) Linear correlation between PD-L1+ TANs and PD-1+ CD4+ T cells, PD-1+ CD8+ T cells, or PD-1+ γδT cells. Each dot represents an independent data point as determined by flow cytometry. ns = p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: For flow cytometry, cell suspensions from human peripheral blood and tissue samples were stained with following antibodies cocktail at 4°C for 40 mins: Percp-Cy TM 5.5 mouse anti-human CD45 (564,105, Clone HI30, BD Pharmingen TM , San Diego, USA), BV510 mouse anti-human CD3 (564,713, Clone HIT3α, BD Pharmingen TM , San Diego, USA), APC-R700 mouse anti-human CD4 (564,975, Clone RPA-T4, BD Pharmingen TM , San Diego, USA), APC-Cy TM 7 mouse anti-human CD8 (557,834, Clone SK1, BD Pharmingen TM , San Diego, USA), PE mouse anti-human TCR γδ (555,717, Clone B1, BD Pharmingen TM , San Diego, USA), Alexa Fluor ® 647 mouse anti-human CD66b (561,645, Clone G10F5, BD Pharmingen TM , San Diego, USA), BV421 mouse anti-human PD-1 (564,323, Clone MIH4, BD Horizon TM , San Diego, USA), PE-Cy TM 7 mouse anti-human CD274 (558,017, Clone MIH1, BD Pharmingen TM , San Diego, USA).

Techniques: Expressing, Flow Cytometry