Journal: Cell Reports Medicine

Article Title: Merkel cell polyomavirus-specific and CD39 + CLA + CD8 T cells as blood-based predictive biomarkers for PD-1 blockade in Merkel cell carcinoma

doi: 10.1016/j.xcrm.2023.101390

Figure Lengend Snippet:

Article Snippet: 114 - CD56 (clone: REA196) , Miltenyi Biotec , Cat# 130-108-016, RRID: AB_2658728.

Techniques: Recombinant, Control, Sequencing, RNA Sequencing, Software

Journal: International Journal of Molecular Sciences

Article Title: Glycan Structures in Osteosarcoma as Targets for Lectin-Based Chimeric Antigen Receptor Immunotherapy

doi: 10.3390/ijms25105344

Figure Lengend Snippet: CD301-CARs display specific cytotoxicity against osteosarcoma cell lines. ( A ) Schematic representation of the CD301 LEC-CAR: ( B ) Expression of CD301 CAR at the cell surface of NK92 cells. NK92 cells were retrovirally transduced and sorted based on the GFP expression. The expression of the CD301 CAR construct was measured by flow cytometry using an antibody specific to the CD301 CRD. Color denotes areas of high and low population density. ( C ) Detection of CD301 ligands on target cells: osteosarcoma cell lines were stained with fluorescently labeled recombinant CD301 and analyzed by flow cytometry. ( D ) Cytotoxicity assay: NK92 cells expressing the CD301-CAR were cocultured for 3 h with calcein-labeled target cells with an effector-to-target ratio of 3:1 and 5:1, respectively, and measured as quadruplicates. Columns represent the median. Error bars show standard deviation. p < 0.01 is indicated by **, respectively.

Article Snippet: The degranulation of NK92 CAR and wild-type NK92 cells was induced upon interaction with target cells at an E:T ratio of 1:1 for 4 h at 37 °C and was assessed by measuring the expression of CD107a with a PE/Cy7-labeled anti-CD107a antibody (Biolegend, #328617) at the surface of FITC-labeled anti-CD56-stained NK92 cells (Miltenyi, #130-114-740).

Techniques: Expressing, Construct, Flow Cytometry, Staining, Labeling, Recombinant, Cytotoxicity Assay, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Glycan Structures in Osteosarcoma as Targets for Lectin-Based Chimeric Antigen Receptor Immunotherapy

doi: 10.3390/ijms25105344

Figure Lengend Snippet: Lytic activity of CD301-CAR-expressing NK92 cells correlates with interferon-gamma secretion and increased degranulation. ( A ) CAR expression leads to enhanced interferon-gamma secretion of NK92 cells upon engagement with osteosarcoma cell lines. ( B ) CAR expression leads to enhanced degranulation of NK92 cells upon engagement with osteosarcoma cells. Columns represent the mean of three independent experiments measured in triplicates. Error bars show the standard deviation of the mean. p < 0.05, p < 0.01, and p < 0.001 are indicated by *, **, or *** respectively.

Article Snippet: The degranulation of NK92 CAR and wild-type NK92 cells was induced upon interaction with target cells at an E:T ratio of 1:1 for 4 h at 37 °C and was assessed by measuring the expression of CD107a with a PE/Cy7-labeled anti-CD107a antibody (Biolegend, #328617) at the surface of FITC-labeled anti-CD56-stained NK92 cells (Miltenyi, #130-114-740).

Techniques: Activity Assay, Expressing, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Glycan Structures in Osteosarcoma as Targets for Lectin-Based Chimeric Antigen Receptor Immunotherapy

doi: 10.3390/ijms25105344

Figure Lengend Snippet: Combination of immune checkpoint inhibition with CD301 CAR immunotherapy. ( A ) Flow cytometry analysis showing the surface expression of the TIGIT and PVRIG on NK92 cells. Respective antibody staining is shown as a red histogram, and the isotype control and unstained control are depicted as blue and gray histograms. ( B ) Target cell expression of PVR and PVRL2. Expression was analyzed by flow cytometry. The binding of anti-PVR and anti-PVRL2 antibodies are shown as orange histograms. The isotype and unstained control are shown as blue and gray histograms. ( C ) Cytotoxicity assay: NK92 cells expressing the CD301-CAR were cocultured for 3 h with calcein-labeled targets with an effector target ratio of 3:1, respectively, in the presence or absence of different amounts of the inhibitory anti-TIGIT antibody. Measurements were performed as quadruplicates. Columns represent the mean of three independent experiments. Error bars show the standard deviation of the mean. p < 0.05 and p < 0.01 are indicated by * and **, respectively.

Article Snippet: The degranulation of NK92 CAR and wild-type NK92 cells was induced upon interaction with target cells at an E:T ratio of 1:1 for 4 h at 37 °C and was assessed by measuring the expression of CD107a with a PE/Cy7-labeled anti-CD107a antibody (Biolegend, #328617) at the surface of FITC-labeled anti-CD56-stained NK92 cells (Miltenyi, #130-114-740).

Techniques: Inhibition, Flow Cytometry, Expressing, Staining, Control, Binding Assay, Cytotoxicity Assay, Labeling, Standard Deviation