Journal: iScience

Article Title: Single high-fat challenge and trained innate immunity: A randomized controlled cross-over trial

doi: 10.1016/j.isci.2024.111103

Figure Lengend Snippet:

Article Snippet: Using the lysis-no-wash strategy (BD Pharm Lyse lysing buffer, Becton Dickinson), 50 μL EDTA blood was stained with monoclonal antibodies targeting CD45 (anti-human CD45 KromeOrange, Clone J33, Beckman Coulter), CD16 (anti-human CD16 FITC, Clone CD16, eBioscience), CD14 (anti-human CD14 PECy7, Clone 61D3, eBioscience), CD3 (anti-human CD3 APC-Alexa750, clone UCTH1, Beckman Coulter), CD56 (anti-human CD56 APC, Clone N901, Beckman Coulter), HLA-DR (anti-human HLA-DR PE, Clone immu-357, Beckman Coulter), CD11b (anti-human CD11b BV785, Clone ICRF44, Biolegend), CCR2 (anti-human CD192 (CCR2) BV421, Clone 48607, Becton Dickinson) and CD36 (anti-human CD36 PC5.5, Clone 5–271, Biolegend).

Techniques: Recombinant, Saline, Enzyme-linked Immunosorbent Assay, Software, Flow Cytometry

Journal: Biomedicines

Article Title: Potential Role of Circulating PD-L1 + Leukocytes as a Predictor of Response to Anti-PD-(L)1 Therapy in NSCLC Patients

doi: 10.3390/biomedicines12050958

Figure Lengend Snippet: Pretreatment percentages of PD-L1 + leukocytes and platelets in NSCLC patients based on anti-PD-(L)1 response. Comparison of PD-L1 + ( A ) CD14 + cells, ( B ) neutrophils, ( C ) NK cells, ( D ) CD4 + T cells, ( E ) CD8 + T cells, and ( F ) platelets among PD (n = 55; except for platelets, n = 34), SD (n = 24; except for platelets, n = 17) and R patients (n = 34; except for platelets, n = 22). ( G – I ) Spearman correlations between PD-L1 + CD14 + cells, PD-L1 + neutrophils, and PD-L1 + platelets. ( J ) Comparison of the plasma sPD-L1 concentrations between PD, SD, and R patients. The white dots represent patients with complete response and green dots represent patients with partial response. Statistical analysis between three groups of patients was performed using the Kruskal–Wallis test with Dunn’s correction. p -values are shown in graphs (*) p < 0.05; (**) p < 0.01. n.s. is not significant.

Article Snippet: One hundred microliters of whole blood were stained with anti-CD3-PECy7 (BioLegend, San Diego, CA, USA; clone HIT3a), anti-CD8-PECy5 (BioLegend; clone SK1), anti-PD-L1-PE (BioLegend; clone 29E.2A3), anti-CD14-PECy7 (BD Bioscience, San Jose, CA, USA; clone M5E2), anti-CD41a-FITC (Immunotools, Friesoythe, Germany; clone HIP8), and anti-CD62P-APC (Immunotools; clone HI62P).

Techniques: Comparison

Journal: Biomedicines

Article Title: Potential Role of Circulating PD-L1 + Leukocytes as a Predictor of Response to Anti-PD-(L)1 Therapy in NSCLC Patients

doi: 10.3390/biomedicines12050958

Figure Lengend Snippet: Kinetics of PD-L1 + CD14 + cells, PD-L1 + neutrophils, and PD-L1 + platelets in NSCLC patients based on anti-PD-(L)1 response. Percentages of ( A ) PD-L1 + CD14 + cells, ( B ) PD-L1 + neutrophils, and ( C ) PD-L1 + platelets during 24 weeks of anti-PD-(L)1 treatment in PD (red line), SD (blue line), and R patients (green line). p -values from one-way ANOVAs for each of the three groups are displayed on the right side of each graph, and significant differences between successive time intervals are analyzed by Tukey post hoc comparisons and marked with a line. Changes (>10% decrease (black), no significant change (with less than a 10% change) (light grey), and >10% increase (dark grey)) in the percentages of ( D ) CD14 + cells, ( E ) neutrophils, and ( F ) platelets in the first 4–6 weeks of anti-PD-(L)1 treatment. (*) p < 0.05; n.s. is not significant.

Article Snippet: One hundred microliters of whole blood were stained with anti-CD3-PECy7 (BioLegend, San Diego, CA, USA; clone HIT3a), anti-CD8-PECy5 (BioLegend; clone SK1), anti-PD-L1-PE (BioLegend; clone 29E.2A3), anti-CD14-PECy7 (BD Bioscience, San Jose, CA, USA; clone M5E2), anti-CD41a-FITC (Immunotools, Friesoythe, Germany; clone HIP8), and anti-CD62P-APC (Immunotools; clone HI62P).

Techniques:

Journal: Biomedicines

Article Title: Potential Role of Circulating PD-L1 + Leukocytes as a Predictor of Response to Anti-PD-(L)1 Therapy in NSCLC Patients

doi: 10.3390/biomedicines12050958

Figure Lengend Snippet: Kinetics of CD14 + cells and neutrophils with or without bound platelets and their PD-L1 expression in NSCLC patients based on anti-PD-(L)1 response. ( A , B ) CD14 + cells and ( C , D ) neutrophils with bound platelets and their evolution throughout 22–24 weeks of treatment in PD (n = 55), SD (n = 24), and R patients (n = 34). White dots represent patients with complete response and green dots patients with partial response. Kinetics of PD-L1 + ( E ) CD14 + cells and ( F ) neutrophils without bound platelets during 24 weeks of anti-PD-(L)1 therapy. p -values from one-way ANOVAs for each of the three groups are displayed on the right side of each graph, and significant differences between successive time intervals are analyzed by the Tukey post hoc comparisons and marked with a line. ( G ) Comparison of pretreatment baseline percentages of PD-L1 + CD14 + cells and PD-L1 + neutrophils with or without bound platelets among NSCLC classified according to their response to anti-PD-(L)1 therapy. Statistical analyses between three groups of patients were performed using Kruskal–Wallis test with Dunn’s correction (showed in the table). (*) p < 0.05; n.s. is not significant.

Article Snippet: One hundred microliters of whole blood were stained with anti-CD3-PECy7 (BioLegend, San Diego, CA, USA; clone HIT3a), anti-CD8-PECy5 (BioLegend; clone SK1), anti-PD-L1-PE (BioLegend; clone 29E.2A3), anti-CD14-PECy7 (BD Bioscience, San Jose, CA, USA; clone M5E2), anti-CD41a-FITC (Immunotools, Friesoythe, Germany; clone HIP8), and anti-CD62P-APC (Immunotools; clone HI62P).

Techniques: Expressing, Comparison

Journal: Biomedicines

Article Title: Potential Role of Circulating PD-L1 + Leukocytes as a Predictor of Response to Anti-PD-(L)1 Therapy in NSCLC Patients

doi: 10.3390/biomedicines12050958

Figure Lengend Snippet: Multinomial logistic regression analysis and pretreatment PD-L1 + CD14 + cells/IL-17 ratio. ( A ) The reference group is R patients (PD vs. R in red; SD vs. R in blue). Dots represent the odds ratio and the error bars represent the 95% CI. ( B ) Ratio between the percentage of PD-L1 + CD14 + cells and plasma IL-17 concentration before starting anti-PD-(L)1 therapy. The Kruskal–Wallis test with Dunn’s correction was used to compare the baseline ratio between studied groups. p -values are shown in graphs (*) p < 0.05; (**) p < 0.01. CI, confidence interval.

Article Snippet: One hundred microliters of whole blood were stained with anti-CD3-PECy7 (BioLegend, San Diego, CA, USA; clone HIT3a), anti-CD8-PECy5 (BioLegend; clone SK1), anti-PD-L1-PE (BioLegend; clone 29E.2A3), anti-CD14-PECy7 (BD Bioscience, San Jose, CA, USA; clone M5E2), anti-CD41a-FITC (Immunotools, Friesoythe, Germany; clone HIP8), and anti-CD62P-APC (Immunotools; clone HI62P).

Techniques: Concentration Assay

Journal: Nature Communications

Article Title: Deciphering the spatial landscape and plasticity of immunosuppressive fibroblasts in breast cancer

doi: 10.1038/s41467-024-47068-z

Figure Lengend Snippet: A CellChat dominant sender and receiver plot showing incoming and outgoing interaction strength for each cell type identified in the BC atlas (73,426 cells from 43 patients). The size of each circle corresponds to the total number of significant interactions, colored per cell type. B Scatter plots with linear regression lines showing correlations in the content of FAP+ CAF clusters quantified by flow cytometry in BC ( N = 87 patients). p values from two-sided Pearson’s correlation test. C Same as ( B ) analyzing correlations between the content in ECM-myCAF, TGFβ-myCAF, Wound-myCAF and Detox-iCAF with TREM2+ or FOLR2+ macrophages in BC ( N = 25). D Bar plot showing the percentages (%) of migration of CD14+ monocytes after 6 h of transwell co-culture with FAP+ CAF clusters. Data are mean ± SEM ( n = 4 independent experiments). p values from two-sided Student’s t test. E % of CD14+ CD16+ myeloid cells among total CD14+ monocytes after 24 h of co-culture with FAP+ CAF clusters (Detailed gating strategy in Supplementary Fig. ). Data are mean ± SEM ( n = 9). p values from two-sided Student’s t test. F Same as ( E ) for TREM2+ macrophages. p values from two-sided Student’s t test. G Same as ( E ) for FOLR2+ macrophages. p values from two-sided Mann–Whitney test. H % of FOXP3+ regulatory T cells among CD4+ CD25+ T lymphocytes after 16 h of co-culture with FAP+ CAF clusters (Detailed gating strategy in Supplementary Fig. ). Data are mean ± SEM ( n = 8). p values from two-sided Mann–Whitney test. I Same as ( H ) for the % of PD-1+ FOXP3+ T lymphocytes. p values from two-sided Mann–Whitney test. J Same as ( H ) for the % of CTLA-4+ FOXP3+ T lymphocytes. p values from two-sided Mann–Whitney test. K % of Perforin+ among total CD16+ NK cells after 24 h of co-culture with FAP+ CAF clusters (Detailed gating strategy in Supplementary Fig. ). Data are mean ± SEM ( n = 7). p values from two-sided Student’s t test. L Same as ( K ) for Granzyme B+ NK cells. p values from two-sided Student’s t test. M Same as ( K ) for CD16+ CD56 high NK cells. p values from two-sided Student’s t test. N Same as ( K ) for NKG2A+ NK cells. p values from two-sided Student’s t test. Source data are provided as a Source Data file.

Article Snippet: Isotype control antibodies for macrophages subsets were used: iso-anti-CD16-BV650 (BV650 Mouse IgG1 κ Isotype Control, 1:50, BD Biosciences, #563231), iso-anti-CD56-BUV395 (BUV395 Mouse IgG2b κ Isotype Control; 1:50, BD Biosciences, #563558), iso-anti-CD14-Pecy7 (PE-Cy7 Mouse IgG2a κ Isotype Control, 1:50, BD Biosciences, #557907), iso-anti-FOLR2-PEC (PE Mouse IgG1 κ Isotype Ctrl Antibody; 1:100, Biolegend, #400112) and iso-anti-TREM2-APC (Rat IgG2B Isotype Control, 1:50, Novus Biologicals, #MAB0061).

Techniques: Flow Cytometry, Migration, Co-Culture Assay, MANN-WHITNEY