Journal: Journal of Translational Medicine

Article Title: First-line chemoimmunotherapy in metastatic breast carcinoma: combination of paclitaxel and IMP321 (LAG-3Ig) enhances immune responses and antitumor activity

doi: 10.1186/1479-5876-8-71

Figure Lengend Snippet: IMP321 increases the numbers of monocytes, NK and activated CD8 T cells in blood (panel A) . Fresh blood samples were collected pre-dosing, at D1, D85 and D170 and directly stained with fluorochrome-conjugated antibodies in tubes containing a precise number of fluorescent control beads. The results show the mean ± sd of the absolute numbers of CD45 + CD14 + (monocytes), CD3 - CD56 + (NK cells) and CD38 + HLA-DR + CD8 + (activated CD8 + cells) cells. The paired non-parametric Wilcoxon signed rank test was used to compare increases observed between D85 or D170 and D1. When significant (< 0.05), p values are indicated. IMP321 increases the percentages of dendritic cells and cytotoxic CD45RA + Effector-Memory CD8 + T cells (EMRA) in PBMCs (panel B). PBMCs cells collected pre-dosing, at D1, D85 and D170 were isolated and stained with fluorochrome-conjugated antibodies and analyzed by flow cytometry. The results show the mean ± sd of the percentages of plasmacytoid dendritic cells (pDC, CD45 + CD14 - CD16 - HLA-DR + CD123 + CD11c - ) and myeloid dendritic cells (mDC, CD45 + CD14 - CD16 - HLA-DR + CD123 - CD11c + ) in CD45 + cells, as well as the percentages of CD45RA + CD45RO - CD62L - in the CD8 + T cell population (CD45RA + EM CD8 T cells or EMRA). The significant Wilcoxon p values are indicated.

Article Snippet: Blood samples were collected pre-dosing at D1, D85 and D170 and directly stained with BD Multitest CD8-FITC/CD38-PE/CD3-PerCP/HLA-DR-APC, with BD Multitest 6-color TBNK (CD3-FITC/CD16-PE+CD56-PE/CD45-PerCPCy5.5/CD4PE-Cy7/CD19-APC/CD8-APC-Cy7), BD Simultest LeucoGate (CD45-FITC/CD14-PE) in tubes containing a precise number of fluorescent control beads (BD Trucount™tubes, BD Biosciences).

Techniques: Staining, Isolation, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: First-line chemoimmunotherapy in metastatic breast carcinoma: combination of paclitaxel and IMP321 (LAG-3Ig) enhances immune responses and antitumor activity

doi: 10.1186/1479-5876-8-71

Figure Lengend Snippet: IMP321 increases the expression of activation markers on blood monocytes . Blood samples were collected pre-dosing, at D1, D85 and D170 and directly stained with fluorochrome-conjugated CD45, CD14, anti-HLA-DR and CD11a, CD11b, CD16, CD35, CD54, CD64, CD80 or CD86 antibodies in tubes containing a precise number of fluorescent control beads. The expression of activation markers on monocytes was directly proportional to the cell-bound fluorescence. The results shown in panel A are the mean ± sd after normalization of the cell-bound fluorescence against the fluorescence of control beads. Statistically significant increases between D85 or D170 and D1 are analyzed using Wilcoxon signed rank test and significant p values (< 0.05) are shown. In panel B, the percentage of patients showing increases in the expression of the indicated numbers of activation markers at D85 or D170 compared to the baseline at D1 was calculated. The number of markers (n) displaying an increase by at least 50% was calculated for each patient in the 1.25 mg (7 patients) and 6.25 mg (12 patients) groups. The pie charts represent the percentages of patients with increases in the indicated number of markers.

Article Snippet: Blood samples were collected pre-dosing at D1, D85 and D170 and directly stained with BD Multitest CD8-FITC/CD38-PE/CD3-PerCP/HLA-DR-APC, with BD Multitest 6-color TBNK (CD3-FITC/CD16-PE+CD56-PE/CD45-PerCPCy5.5/CD4PE-Cy7/CD19-APC/CD8-APC-Cy7), BD Simultest LeucoGate (CD45-FITC/CD14-PE) in tubes containing a precise number of fluorescent control beads (BD Trucount™tubes, BD Biosciences).

Techniques: Expressing, Activation Assay, Staining, Fluorescence

Journal: Cell

Article Title: Passive Transfer of Vaccine-Elicited Antibodies Protects Against SIV in Rhesus Macaques

doi: 10.1016/j.cell.2020.08.033

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: APC-Cy7 Mouse Anti-Human CD16 , BD Biosciences , CAT#557758 RRID:AB_396853.

Techniques: Purification, Recombinant, Labeling, Flow Cytometry, RNA Extraction, Software, Luminex, Magnetic Beads

Journal: Frontiers in Immunology

Article Title: Long-term use of rituximab increases T cell count in MS patients

doi: 10.3389/fimmu.2024.1412668

Figure Lengend Snippet: The effect of RTX on the B cell population. (A) The relative change in number of CD19 + cells/μL after the first RTX administration comparing two doses. (B) CD19 + cell depletion after the first administration of RTX comparing two doses. (C, D) Number of CD19 + cells/μL 0-200 and 201-300 days after the administration of RTX, respectively. *p=<0.05, **p=<0.01, ***p=<0.001, ****p=<0.0001. ns, not significant.

Article Snippet: After January 2023 differential counts were performed with a BD Multitest™ 6-color TBNK Reagent and BD Trucount™ (CD3-FITC, CD16-PE, CD56-PE, CD45-PerCP-Cy5.5, CD4-PE-Cy7, CD19-APC, CD8-APC-Cy7) and Trucount Absolute Counting Tubes and analyzed on a FACSLyric instrument (BD Bioscieneces).

Techniques:

Journal: Frontiers in Immunology

Article Title: Long-term use of rituximab increases T cell count in MS patients

doi: 10.3389/fimmu.2024.1412668

Figure Lengend Snippet: Short term effect of RTX on CD3 + cells. (A) The number of T cells (CD3 + cells/μL) at baseline and after the first RTX administration regardless of dosage (n=146), difference not significant. Wilcoxon test. (B) The relative change in the number of T cells (CD3 + cells/μL) after the first RTX administration comparing the two doses 1000mg (n=56) and 500mg (n=90), difference not significant. Mann-Whitney test. (C) The number of T cells (CD3 + cells/μL) at baseline and after the last RTX administration (3-8 RTX administrations) regardless of dosage (n=97), p=0.0084, Wilcoxon test. **p=<0.01. ns, not significant.

Article Snippet: After January 2023 differential counts were performed with a BD Multitest™ 6-color TBNK Reagent and BD Trucount™ (CD3-FITC, CD16-PE, CD56-PE, CD45-PerCP-Cy5.5, CD4-PE-Cy7, CD19-APC, CD8-APC-Cy7) and Trucount Absolute Counting Tubes and analyzed on a FACSLyric instrument (BD Bioscieneces).

Techniques: MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Long-term use of rituximab increases T cell count in MS patients

doi: 10.3389/fimmu.2024.1412668

Figure Lengend Snippet: Long-term effects of RTX on T cells. (A) Frequency of CD3 + cells/μL. (B) CD4/CD8 ratio. (C) Frequency of CD4 + cells/μL. (D) Frequency of CD8 + cells/μL. (E) Percentage of CD4 + /HLA-DR + /CD38 + (activated) cells. (F) Percentage of CD4 + /CCR7 + /CD45RA + (näive) cells. (G) Percentage of CD4 + /CCR7 neg /CD45RA neg (effector) cells. (H) Percentage of CD8 + /HLA-DR + /CD38 + (activated) cells. (I) Percentage of CD8 + /CCR7 + /CD45RA + (näive) cells. (J) Percentage of CD8 + /CCR7 neg /CD45RA neg (effector) cells. *p=<0.05, **p=<0.01, ***p=<0.001, ****p=<0.0001. ns, not significant.

Article Snippet: After January 2023 differential counts were performed with a BD Multitest™ 6-color TBNK Reagent and BD Trucount™ (CD3-FITC, CD16-PE, CD56-PE, CD45-PerCP-Cy5.5, CD4-PE-Cy7, CD19-APC, CD8-APC-Cy7) and Trucount Absolute Counting Tubes and analyzed on a FACSLyric instrument (BD Bioscieneces).

Techniques:

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