cd39 fitc  (Miltenyi Biotec)

Journal: Purinergic Signalling

Article Title: Characterization of purinergic signaling in tumor-infiltrating lymphocytes from lower- and high-grade gliomas

doi: 10.1007/s11302-023-09931-4

Figure Lengend Snippet: Glioma-derived primary cell culture. Representative images of primary cultures derived from different glioma grades. Almost all cells were GFAP positive (some with very low expression, green). DAPI was used to stain the nucleus (blue). Scale bar = 100 µm (A-C). No difference between LGG and HGG was observed in primary cell culture regarding CD38, CD39, and CD73 ectoenzymes protein and mRNA expression (D-K). Cells were incubated with ATP 100 μM for 15, 30, and 60 min, and nucleotides and nucleosides metabolism was evaluated by HPLC. LGG cell cultures show a significant increase in ADO production compared to HGG (L-O). The experiment was performed in triplicate using primary cell cultures derived from eleven patients with different glioma grades. Data is shown as the mean ± SD

Article Snippet: Flow cytometry of tumor-infiltrated leukocytes and peripheral blood immune cells After being isolated, tumor-infiltrated leukocytes and PBMC were blocked for non-specific binding using a FACS buffer (Phosphate Buffered Saline + 2% FBS) and stained with the following panel: CD45-V500 (clone: HI30, cat. 560,777) / CD4-Pacific Blue (clone: RPA-T4, cat. 558,116) / CD39-FITC (clone: TU66, cat. 561,444) / CD73-PE (clone: AD2, cat. 550,257)/ CD19-PE-Cy7 (clone: SJ25C1, cat. 557,835) / CD8-APC (clone: RPA-T8, cat. 555,369) / CD38-APCH7 (clone: HB7, cat. 656,646) (BD Biosciences, USA) for 30 min at 4ºC.

Techniques: Derivative Assay, Cell Culture, Expressing, Staining, Incubation

Journal: Purinergic Signalling

Article Title: Characterization of purinergic signaling in tumor-infiltrating lymphocytes from lower- and high-grade gliomas

doi: 10.1007/s11302-023-09931-4

Figure Lengend Snippet: CD39-CD73 canonical axis in glioma-infiltrating immune cells compared to matched peripheral blood in LGG, HGG IDH-Mut, and HGG IDH-WT. Percentage of canonical ectoenzymes CD39, CD73, and CD39 + CD73 + expression on total lymphocytes (A), CD4 + T cell (B), CD8 + T cell (C), CD19 + B cell (D), and NK (E). Representative histograms highlighting the difference in CD39 and CD73 expression between tumor-infiltrating lymphocytes and peripheral blood in all analyzed groups (F – H). Data represents a combination of experiments involving individual patients and is displayed as the mean ± SD

Article Snippet: Flow cytometry of tumor-infiltrated leukocytes and peripheral blood immune cells After being isolated, tumor-infiltrated leukocytes and PBMC were blocked for non-specific binding using a FACS buffer (Phosphate Buffered Saline + 2% FBS) and stained with the following panel: CD45-V500 (clone: HI30, cat. 560,777) / CD4-Pacific Blue (clone: RPA-T4, cat. 558,116) / CD39-FITC (clone: TU66, cat. 561,444) / CD73-PE (clone: AD2, cat. 550,257)/ CD19-PE-Cy7 (clone: SJ25C1, cat. 557,835) / CD8-APC (clone: RPA-T8, cat. 555,369) / CD38-APCH7 (clone: HB7, cat. 656,646) (BD Biosciences, USA) for 30 min at 4ºC.

Techniques: Expressing

Journal: Purinergic Signalling

Article Title: Characterization of purinergic signaling in tumor-infiltrating lymphocytes from lower- and high-grade gliomas

doi: 10.1007/s11302-023-09931-4

Figure Lengend Snippet: Purinergic ectoenzymes (A – E; J) and adenosinergic receptors (F – I) gene expression were evaluated by qRT-PCR and HGG tumor samples were compared to LGG. Graphs show the correlation between the percentage of CD38’ tumor-infiltrating lymphocytes and CD38 mRNA expression from tumor sample (K), the percentage of CD39’ tumor-infiltrating lymphocytes and ENTPD1 mRNA expression from tumor sample (L), and the percentage of CD73 tumor-infiltrating lymphocytes and NT5E mRNA expression from tumor sample (M), regardless the tumor grade. Kaplan–Meier analysis showing the influence of CD38, ENTPD1, and NT5E genes expressions on HGG and LGG OS (N – S). Data represent a combination of experiments involving individual patients and are displayed as a single value by the Spearman test

Article Snippet: Flow cytometry of tumor-infiltrated leukocytes and peripheral blood immune cells After being isolated, tumor-infiltrated leukocytes and PBMC were blocked for non-specific binding using a FACS buffer (Phosphate Buffered Saline + 2% FBS) and stained with the following panel: CD45-V500 (clone: HI30, cat. 560,777) / CD4-Pacific Blue (clone: RPA-T4, cat. 558,116) / CD39-FITC (clone: TU66, cat. 561,444) / CD73-PE (clone: AD2, cat. 550,257)/ CD19-PE-Cy7 (clone: SJ25C1, cat. 557,835) / CD8-APC (clone: RPA-T8, cat. 555,369) / CD38-APCH7 (clone: HB7, cat. 656,646) (BD Biosciences, USA) for 30 min at 4ºC.

Techniques: Expressing, Quantitative RT-PCR

Journal: Biomolecules

Article Title: Altered CD39 and CD73 Expression in Rheumatoid Arthritis: Implications for Disease Activity and Treatment Response

doi: 10.3390/biom14010001

Figure Lengend Snippet: Flow cytometric analysis of CD39 and CD73 on monocytes, CD8, CD4, Treg, B, and NK cells from peripheral blood of untreated RA patients (RA) and healthy controls (C). Representative dot plots for surface expression of CD39 and CD73 and frequencies of different subpopulations (CD39 + CD73 + , CD39 + CD73 − , CD39 − CD73 + , and CD39 − CD73 − ). Data are expressed as means and SD (standard deviation) of the percentage of the corresponding population (CD4 + T cells, CD8 + T cells, B cells, NK, lymphocytes, and monocytes). In the case of Treg (CD4 + CD25 high CD127 low ) the percentage is expressed for total CD4 + T cells. An unpaired t -test was used to compare the percentages in the RA and C groups.

Article Snippet: The PBMCs were stained with the following monoclonal antibodies: anti-human CD3-Viogreen (Miltenyi Biotec GmBh), CD39-Fitc and CD73-PE (BD Biosciences), and CD8-PECy7 (Biolegend).

Techniques: Expressing, Standard Deviation

Journal: Biomolecules

Article Title: Altered CD39 and CD73 Expression in Rheumatoid Arthritis: Implications for Disease Activity and Treatment Response

doi: 10.3390/biom14010001

Figure Lengend Snippet: Correlations between some subsets and clinical parameters. Correlation plots corresponding to relationships between disease activity indexes and the percentages of CD8 + CD39 − CD73 + , CD8 + CD39 − CD73 − , Treg CD39 + CD73 + , Treg CD39 − CD73 − or B CD39 − CD73 − subsets in RA patients at baseline. Only the plots with a tendency line presented a significative correlation applying Pearson’s test.

Article Snippet: The PBMCs were stained with the following monoclonal antibodies: anti-human CD3-Viogreen (Miltenyi Biotec GmBh), CD39-Fitc and CD73-PE (BD Biosciences), and CD8-PECy7 (Biolegend).

Techniques: Activity Assay

Journal: Biomolecules

Article Title: Altered CD39 and CD73 Expression in Rheumatoid Arthritis: Implications for Disease Activity and Treatment Response

doi: 10.3390/biom14010001

Figure Lengend Snippet: Correlations between subsets and clinical parameters ( a ) Correlograms corresponding to R-RA (n = 20) and NR-RA (n = 10) patients. Squares marked with a black border in correlogram indicate significant correlations observed in R-RA, but not in NR-RA patients. ( b ) Correlation plots illustrating the relationship between disease activity indexes and the percentage of Treg CD39 + CD73 − , CD8 + CD39 − CD73 + , CD8 + CD39 − CD73 − , B CD39 − CD73 − , B CD39 − CD73 + or B CD39 + CD73 − subsets in R-RA patients at baseline. p = 0.05 *, p = 0.01 ** and p = 0.001 ***.

Article Snippet: The PBMCs were stained with the following monoclonal antibodies: anti-human CD3-Viogreen (Miltenyi Biotec GmBh), CD39-Fitc and CD73-PE (BD Biosciences), and CD8-PECy7 (Biolegend).

Techniques: Activity Assay

Journal: Biomolecules

Article Title: Altered CD39 and CD73 Expression in Rheumatoid Arthritis: Implications for Disease Activity and Treatment Response

doi: 10.3390/biom14010001

Figure Lengend Snippet: Effects of stimulation on CD39 and CD73 expression. ( a ) Representative dot plots for surface expression of CD39 and CD73 on CD4 + T and CD8 + T cells on PBMCs from Cs before culture and after 72 h cultured without beads (medium) or with beads (medium + beads). Additionally, PBMCs from Cs were cultured with IL-6, both without beads (IL-6) and with beads (IL-6 + beads). ( b ) Percentages of the four subsets of CD4 + T and CD8 + T cells from PBMCs cultured for 72 h either with anti-CD3/CD28 beads (empty dots) or without beads (full dots). ( c ) Percentages of subsets cultured under the same conditions, but with IL-6. The double-negative subset in CD4 + T and CD8 + T cells as well as the CD8 + CD39 − CD73 + subset are represented in the right axes. The Wilcoxon signed-rank test was applied in comparisons.

Article Snippet: The PBMCs were stained with the following monoclonal antibodies: anti-human CD3-Viogreen (Miltenyi Biotec GmBh), CD39-Fitc and CD73-PE (BD Biosciences), and CD8-PECy7 (Biolegend).

Techniques: Expressing, Cell Culture

Journal: Biomolecules

Article Title: Altered CD39 and CD73 Expression in Rheumatoid Arthritis: Implications for Disease Activity and Treatment Response

doi: 10.3390/biom14010001

Figure Lengend Snippet: Effects of stimulation on CD39 and CD73 expression, and IFNγ production in CD4 + T and CD8 + T cells. The PBMCs from Cs (gray symbols) and RA patients (black symbols) were cultured for 72 h with beads and with (triangles) or without IL-6 (dots). The percentages of IFNγ-producing cells in CD4 + T (CD4 + IFN + ) and CD8 + T cells (CD8 + IFN + ) were categorized into two groups based on their expression of CD39 (CD39 + and CD39 − ) or CD73 (CD73 + and CD73 − ). In all plots, symbols colored in red correspond to NR-RA patients. Paired t -tests were applied in comparisons between subsets from C or RA patients and unpaired t -test to compare equivalent subsets in C with RA patients.

Article Snippet: The PBMCs were stained with the following monoclonal antibodies: anti-human CD3-Viogreen (Miltenyi Biotec GmBh), CD39-Fitc and CD73-PE (BD Biosciences), and CD8-PECy7 (Biolegend).

Techniques: Expressing, Cell Culture