Journal: Journal of experimental & clinical cancer research : CR

Article Title: Combinatorial macrophage induced innate immunotherapy against Ewing sarcoma: Turning "Two Keys" simultaneously.

doi: 10.1186/s13046-024-03093-w

Figure Lengend Snippet: Fig. 5 Characterization of markers for ES-phagocytic macrophages. A Gating strategy for evaluating cell surface protein expression on phagocytic versus non-phagocytic macrophages. In-vitro phagocytosis assay was carried out incubating A673 cells (Non- or DOX- or MAG- or D&M-treated) labeled with CMFDA (detected by FITC channel) and M-CSF Mφ. Cells were harvested and stained with APC-CD11b and PE-CD38 antibodies followed by flow cytometry analysis. The viable cells were plotted in FITC (CMFDA) x APC view to visualize cell populations in the in-vitro phagocytosis assay. A gate was drawn to include cells that were FITC positive and APC positive as the phagocytic macrophage population, and another gate was drawn to include cells that were FITC- APC + as the non-phagocytic macrophage population. The FITC + APC + and FITC- APC + cells were then plotted separately in a PE x APC view to visualize expression of CD38 in both cell populations. A quat gate was drawn to visualize APC + PE + as the CD38 + and APC + PE- as the CD38- macrophages. B Comparison of percent of CD14, CD38, CD163, CD206 or CD209 positive and negative cells in the phagocytic and non-phagocytic macrophages incubated with non- or DOX- or MAG- or D&M-treated A673 cells. Macrophages alone (no A673) served as a control. *p < 0.05, **p < 0.01, ***p < 0.001 (Fisher’s Exact test). Shown are results in a representative experiment. The same trend was seen in three independent biological repeats

Article Snippet: Antibodies used include α-CD47 (B6H12, ThermoFisher Scientific, 11–0479-42), α-Calreticulin (ThermoFisher Scientific, PA3-900), α-CD11b (Miltenyi Biotec, 130–110-554), α-CD14 (R&D Systems, FAB3832P-025), α-CD38 (R&D Systems, FAB2404P), α-CD68 (BioLegend, 333,819), α-CD80 (BioLegend, 305,220), α-CD163 (R&D Systems, FAB1607P-025), α-CD206 (R&D Systems, FAB25342P), α-CD209 (R&D Systems, FAB161P-025), Alexa Fluor 488 donkey α-rabbit IgG (Invitrogen, A21206).

Techniques: Expressing, In Vitro, Phagocytosis Assay, Labeling, Staining, Flow Cytometry, Comparison, Incubation, Control

Journal: BMC immunology

Article Title: Cycling and activated CD8 + T lymphocytes and their association with disease severity in influenza patients.

doi: 10.1186/s12865-022-00516-1

Figure Lengend Snippet: Fig. 3 The kinetic changes of cycling and activated T cells during influenza infection were longitudinally characterized. Kinetics of cycling and activated T cells are shown as frequencies, A %Ki67+CD8+ T cells of total CD8+ T cells, B %Ki67+CD4+ T cells of total CD4+ T cells. C %CD38+HLA-DR+CD8+ T cells of total CD8+ T cells, D %CD38+HLA-DR+CD4+ T cells of total CD4+ T cells during the disease course. Statistical significance is determined by unpaired t test. *P < 0.05, **P < 0.01, ns, not significant

Article Snippet: The PBMCs were thawed and analyzed using a panel of antibodies as follows: PE-eFluor 610 anti-human Ki67 (Thermo/eBio); FITC anti-human CD8α (clone Hit8α), PE anti-human CD4 (clone RPA-T4), PE-Cyanine7 antihuman CD38 (clone HB7), violetFluorTM450 anti-human HLA-DR (clone LN3), APC-Cyanine7 anti-human CD27 (clone O323), APC anti-human CD279 (PD-1) (clone J110), PerCP-Cyanine5.5 anti-human CD45RO (clone UCHL1), redFluorTM 710 anti-human CD45RA (clone HI100) (TONBO biosciences).

Techniques: Infection

Journal: Cells

Article Title: The Properties of Proinflammatory Ly6C hi Monocytes Are Differentially Shaped by Parasitic and Bacterial Liver Infections.

doi: 10.3390/cells11162539

Figure Lengend Snippet: Figure 4. Marked differences in expression of surface markers by Ly6Chi monocytes after infection with E. histolytica or L. monocytogenes (A) Heatmap depicting differential expression of mRNA encod- ing selected markers on the surface of liver Ly6Chi monocytes after infection with E. histolytica (E. his) or L. monocytogenes (L. m). (B) Normalized mRNA counts of selected surface marker genes (from transcriptome analysis). (C) Percentage of CD14+, CD38+, CD74+, and CD86+ Ly6Chi monocytes dur- ing the course of infection at the indicated time points post-infection (measured by flow cytometry). (D) Histogram and MFI of CD14+, CD38+, CD74+, and CD86+ Ly6Chi monocytes during the course of infection. Data in C were pooled from three independent experiments. Data in D are representative of one of these three experiments and all data are presented as the mean ± SEM (* p < 0.05; ** p < 0.01; *** p < 0.001, **** p < 0.0001; Mann-Whitney U test).

Article Snippet: The following antibodies were used: CD45.1 BUV395 (A20) (Becton Dickinson); CD11b BV510 (M1/70), CD14 BV421 (1Sa142), CD38 PeCy7 (90), CD45.2 PeCy5 (30-F11), CD74 AF647 (In1/CD74), CD86 AF700 (GL-1), Ly6C FITC/PE (HK1-4), Ly6G BV785 (1A8) (BioLegend); IRF2 AF488 (sc-374327) and mafb PerCP (OTI1E9) (Novus Biologicals).

Techniques: Expressing, Infection, Quantitative Proteomics, Marker, Cytometry, MANN-WHITNEY

Journal: Cells

Article Title: The Properties of Proinflammatory Ly6C hi Monocytes Are Differentially Shaped by Parasitic and Bacterial Liver Infections.

doi: 10.3390/cells11162539

Figure Lengend Snippet: Figure 5. New Ly6Chi monocyte subsets identified according to expression of selected surface markers and IRF2. (A) Gating strategy, histogram, and percentage of IRF2+ Ly6Chi monocytes from livers of naïve mice and from E. histolytica- and L. monocytogenes-infected mice on day 3 post infecion. (B) Gating strategy (exemplary for CD14+ gated cells) based on the FMO control to determine CD14+, CD38+, CD74+, and CD86+ monocytes within the IRF2+ Ly6Chi monocyte population from naïve mice and from both infection models on day 3 post infection. (C) UMAP plots of Ly6Chi

Article Snippet: The following antibodies were used: CD45.1 BUV395 (A20) (Becton Dickinson); CD11b BV510 (M1/70), CD14 BV421 (1Sa142), CD38 PeCy7 (90), CD45.2 PeCy5 (30-F11), CD74 AF647 (In1/CD74), CD86 AF700 (GL-1), Ly6C FITC/PE (HK1-4), Ly6G BV785 (1A8) (BioLegend); IRF2 AF488 (sc-374327) and mafb PerCP (OTI1E9) (Novus Biologicals).

Techniques: Expressing, Infection, Control

Journal: Cells

Article Title: The Properties of Proinflammatory Ly6C hi Monocytes Are Differentially Shaped by Parasitic and Bacterial Liver Infections.

doi: 10.3390/cells11162539

Figure Lengend Snippet: Figure 6. CD38+Ly6Chi monocytes represent the major monocytic source of ROS production and contribute to liver damage following E. histolytica infection. (A) Heat map of differentially regulated genes involved in ROS production and a graph showing fold changes in expression of Ncf1 mRNA in the liver during infection with E. histolytica or L. monocytogenes. (B) Gating scheme and histogram for ROS+ liver Ly6Chi monocytes. Percentage and mean fluorescent intensity (MFI) of ROS+Ly6Chi

Article Snippet: The following antibodies were used: CD45.1 BUV395 (A20) (Becton Dickinson); CD11b BV510 (M1/70), CD14 BV421 (1Sa142), CD38 PeCy7 (90), CD45.2 PeCy5 (30-F11), CD74 AF647 (In1/CD74), CD86 AF700 (GL-1), Ly6C FITC/PE (HK1-4), Ly6G BV785 (1A8) (BioLegend); IRF2 AF488 (sc-374327) and mafb PerCP (OTI1E9) (Novus Biologicals).

Techniques: Infection, Expressing