Journal: Digestive Diseases and Sciences

Article Title: Regulation of the PD-1/PD-L1 Axis and NK Cell Dysfunction by Exosomal miR-552-5p in Gastric Cancer

doi: 10.1007/s10620-024-08536-0

Figure Lengend Snippet: Downregulation of peripheral blood NK cell-activating receptors NKG2D and NKp30 and cytokine factors IFN-γ and TNF-α in gastric cancer. A – C NK cell gating strategy. D and E Flow cytometry analysis comparing the expression of NK cell subsets CD3−/CD56+ and CD3−/CD16+ in GC and HD peripheral blood. F – H Flow cytometry analysis comparing the expression of NK cell receptors NKG2D, NKp30, and NKp46 in GC and HD peripheral blood. I and J ELISA detection of cytokine IFN-γ and TNF-α expression in GC (n = 16) and HD (n = 16) plasma. *P < 0.05; **P < 0.01; ***P < 0.001; ns not significant

Article Snippet: Single-cell suspensions were stained using the following antibodies: anti-human CD3 PE (BioGems, USA), anti-human CD16 PE-Cy7 (BioGems), anti-human CD56 APC (BioGems), anti-human NKG2D (CD314) PE (BioGems), anti-human NKp30(CD337) PE-Cy7 (BD, USA), anti-human NKp46(CD335) APC (BD), and PD-L1 PE (CST, USA).

Techniques: Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay, Clinical Proteomics

Journal: Digestive Diseases and Sciences

Article Title: Regulation of the PD-1/PD-L1 Axis and NK Cell Dysfunction by Exosomal miR-552-5p in Gastric Cancer

doi: 10.1007/s10620-024-08536-0

Figure Lengend Snippet: Correlation of NK cell phenotype and activation receptor gene expression with prognosis in gastric cancer. Kaplan–Meier survival analysis revealed the following associations with poor prognosis in gastric cancer patients: low CD56 expression ( A ), low CD16 expression ( B ), low NKG2D expression ( C ), and low NKp46 expression ( E ). However, no correlation between NKp30 expression and prognosis was observed ( D )

Article Snippet: Single-cell suspensions were stained using the following antibodies: anti-human CD3 PE (BioGems, USA), anti-human CD16 PE-Cy7 (BioGems), anti-human CD56 APC (BioGems), anti-human NKG2D (CD314) PE (BioGems), anti-human NKp30(CD337) PE-Cy7 (BD, USA), anti-human NKp46(CD335) APC (BD), and PD-L1 PE (CST, USA).

Techniques: Activation Assay, Gene Expression, Expressing

Journal: Digestive Diseases and Sciences

Article Title: Regulation of the PD-1/PD-L1 Axis and NK Cell Dysfunction by Exosomal miR-552-5p in Gastric Cancer

doi: 10.1007/s10620-024-08536-0

Figure Lengend Snippet: Correlation of plasma exosomal miR-552-5p with NK cell phenotypes and activated receptors in gastric cancer. A and B Negative correlation between plasma exosomal miR-552-5p expression and the distribution of CD3−/CD56+ and CD3−/CD16+ subpopulations in peripheral blood NK cells of gastric cancer patients. C – E Negative correlation between plasma exosomal miR-552-5p expression and the expression of peripheral blood NK cell-activating receptors NKG2D, NKp30, and NKp46 in gastric cancer

Article Snippet: Single-cell suspensions were stained using the following antibodies: anti-human CD3 PE (BioGems, USA), anti-human CD16 PE-Cy7 (BioGems), anti-human CD56 APC (BioGems), anti-human NKG2D (CD314) PE (BioGems), anti-human NKp30(CD337) PE-Cy7 (BD, USA), anti-human NKp46(CD335) APC (BD), and PD-L1 PE (CST, USA).

Techniques: Clinical Proteomics, Expressing

Journal: JCI Insight

Article Title: Soluble CD13 is a potential mediator of neutrophil-induced thrombogenic inflammation in SARS-CoV-2 infection

doi: 10.1172/jci.insight.184975

Figure Lengend Snippet: ( A ) Reanalysis of single-cell RNA-Seq results generated from circulating human neutrophils revealed that immature neutrophils have higher ANPEP expression. Dot plot of ANPEP , IFN, and neutrophil maturity genes for each neutrophil cluster, showing the average expression level and the percentage of cells expressing the gene in each cluster. ( B ) Gating scheme of flow cytometry analysis on mature and immature neutrophils isolated from whole blood. CD10 + CD16 hi defines mature neutrophils while CD10 – CD16 lo defines immature neutrophils. ( C ) Significantly higher expression of CD13 and lower expression of PAR4 were observed in mature neutrophils compared with immature neutrophils while B1R expression showed similar levels. Data generated from 5–10 healthy controls. Results are expressed as mean ± SD. ** P < 0.01. Significance was determined by Wilcoxon’s test.

Article Snippet: White blood cells in the upper phase were washed with PBS containing 2 mM EDTA and 2% FBS and stained with fluorochrome-conjugated antibodies against human CD3 (PE-Cy7, BioLegend 300420), CD13 (PE, BioLegend 301703), CD14 (APC-Cy7, BioLegend 301820) CD16 (PerCPCy5.5, Tonbo 615-1619-T025), CD19 (BV510, BioLegend 302241), CD45 (APC-Cy7, Tonbo 25-0459-T100), CD56 (PE, BioLegend 362508), CD10 (Super Bright 702, Invitrogen 67-010-642), PAR4 (FITC, Alomone Labs APR-034-F), B1R (APC, LS Bio LS-C275585), MMP14 (APC, R&D Systems, Bio-Techne; FAB9181A), and CD66b (Pacific Blue, BioLegend 305111).

Techniques: RNA Sequencing, Generated, Expressing, Flow Cytometry, Isolation