Journal: Cancers

Article Title: Sperm Protein 17 Expression by Murine Epithelial Ovarian Cancer Cells and Its Impact on Tumor Progression

doi: 10.3390/cancers10080276

Figure Lengend Snippet: Sp17 expression modulates major histocompatibility class (MHC) expression and correlates with immunosuppressive molecules in ID8 cells. ( a ) Classical and non-classical MHC I molecule expression (H2-Db, H2-Kb, H2-M3, Qa-1b, Qa-2, and ribonucleic acid export 1 (rae-1)) in Sp17 + and Sp17 − ID8 cells. Data is shown as histograms of each marker relative expression on Sp17 + (red unshaded) and Sp17 − (blue) ID8 cells, as well as isotype control (orange); ( b ) Immunosuppressive molecules expressions (PD-L1, MHCII, CD39, CD37, CD73, cytotoxic T lymphocyte associated antigen-4 (CTLA-4), tumor necrosis factor receptor II (TNFRII), and STAT3) on gated Sp17 + and Sp17 − ID8 cells. The Sp17 + and Sp17 − ID8 cells (for a) or un-fractionated ID8 cells (for b) were stained with specific antibodies for each immune marker assessed here and analyzed by flow cytometry (see Materials and Methods). Data is shown as histograms for each marker-relative expression in Sp17 + (red) and Sp17 − (blue) ID8 cells, as well as isotype control (orange). The X -axis shows the fluorescence intensity, and Y -axis shows count ( a ) or % expression normalized to mode (as a percent expression relative to each population) ( b ). Results are representative of three independent experiments.

Article Snippet: Cells were stained with fluorochrome-conjugated mAb against PD-L1 (PE), STAT3 (APC), MHC I (biotin), H2-Db (biotin), Qa-1b (biotin), H2-Kb (PE), CTLA-4 (PE), mouse IgG1 (FITC), Streptavidin (Percp) (BD Pharmingen, CA, USA), MHC II (APC-Cy7) (eBioscience, Vienna, Austria), CD39 (PE-Cy7) (eBioscience, Vienna, Austria), CD73 (biotin) (eBioscience, Vienna, Austria), TNFRII (biotin) (eBioscience, Vienna, Austria), and Sp17 antibodies (unconjugated) (SC-365325, Santa Cruz, CA, USA).

Techniques: Expressing, Marker, Staining, Flow Cytometry, Fluorescence

Journal: Cancers

Article Title: Sperm Protein 17 Expression by Murine Epithelial Ovarian Cancer Cells and Its Impact on Tumor Progression

doi: 10.3390/cancers10080276

Figure Lengend Snippet: Sp17 and immune marker expression differs between PD-L1 + MHCII − and PD-L1 − MHCII + populations of ID8 cells. Original (non-clonal selected) ID8 cells were collected and stained for the expression of PD-L1, MHCII, Sp17, STAT3, H2-Db, Qa-1b, and H2-M3. Marker expression between PD-L1 + MHCII − (red) and PD-L1 − MHCII + (blue) populations was analyzed (isotype control, orange). Data is shown as histograms of each marker expression between these two populations.

Article Snippet: Cells were stained with fluorochrome-conjugated mAb against PD-L1 (PE), STAT3 (APC), MHC I (biotin), H2-Db (biotin), Qa-1b (biotin), H2-Kb (PE), CTLA-4 (PE), mouse IgG1 (FITC), Streptavidin (Percp) (BD Pharmingen, CA, USA), MHC II (APC-Cy7) (eBioscience, Vienna, Austria), CD39 (PE-Cy7) (eBioscience, Vienna, Austria), CD73 (biotin) (eBioscience, Vienna, Austria), TNFRII (biotin) (eBioscience, Vienna, Austria), and Sp17 antibodies (unconjugated) (SC-365325, Santa Cruz, CA, USA).

Techniques: Marker, Expressing, Staining

Journal: Cancers

Article Title: Sperm Protein 17 Expression by Murine Epithelial Ovarian Cancer Cells and Its Impact on Tumor Progression

doi: 10.3390/cancers10080276

Figure Lengend Snippet: PD-L1 high MHCII − (Sp17 high ) population of ID8 cells are more Paclitaxel-resistant than PD-L1 − MHCII + (Sp17 low ) populations. ( a ) ID8 cells were FACS-sorted into two populations: PD-L1 high MHCII − and PD-L1 − MHCII + , based on cell surface staining with anti-PD-L1 and anti-MHC II antibodies, which represented the Sp17 high and Sp17 low cells. ( b ) MTT viability assay was used to assess the cytotoxicity of the two populations at multiple time points (24 h, 48 h, and 72 h) and doses (10–1000 nM). Data show the mean percentage of live cells from four samples with standard deviation for each condition. * p < 0.05 (unpaired, two-tailed Student’s t -test).

Article Snippet: Cells were stained with fluorochrome-conjugated mAb against PD-L1 (PE), STAT3 (APC), MHC I (biotin), H2-Db (biotin), Qa-1b (biotin), H2-Kb (PE), CTLA-4 (PE), mouse IgG1 (FITC), Streptavidin (Percp) (BD Pharmingen, CA, USA), MHC II (APC-Cy7) (eBioscience, Vienna, Austria), CD39 (PE-Cy7) (eBioscience, Vienna, Austria), CD73 (biotin) (eBioscience, Vienna, Austria), TNFRII (biotin) (eBioscience, Vienna, Austria), and Sp17 antibodies (unconjugated) (SC-365325, Santa Cruz, CA, USA).

Techniques: Staining, MTT Viability Assay, Standard Deviation, Two Tailed Test

Journal: Molecular imaging and biology

Article Title: An analysis of isoclonal antibody formats suggests a role for measuring PD-L1 with low molecular weight PET radiotracers

doi: 10.1007/s11307-020-01527-3

Figure Lengend Snippet: A summary of the kinetic constants for the C4 minibody, scFv and their respective DFO-conjugates. The binding was assayed against the ectodomain of recombinant human PD-L1 using biolayer interferometry. The data are representative of two independent experiments. In the case of the K D and the K dis for the C4 minibody, the constants approached the limit of detection and the instrument was not capable of reporting error calculations.

Article Snippet: Kinetic constants for the minibody and scFv antibody against human and mouse PD-L1 (Sino Biological Inc.) were determined via biolayer interferometry with an Octet RED384 instrument (ForteBio) using a previously described approach[ 11 ].

Techniques: Binding Assay, Recombinant

Journal: Molecular imaging and biology

Article Title: An analysis of isoclonal antibody formats suggests a role for measuring PD-L1 with low molecular weight PET radiotracers

doi: 10.1007/s11307-020-01527-3

Figure Lengend Snippet: A. DAR and anti-PDL1 immunohistochemistry (co-stained with hematoxylin) of a liver section from a 1 year old female Alb Cre; MYCTg; KRASG12D genetically engineered mouse model of heptacellular carcinoma. Multiple tracer avid foci are detected against the background of normal liver. At right are shown the merged images with the PDL1 immunohistochemistry magnified at 40X in selected fields of view to show the concordance between radiotracer binding and PD-L1 expression in tumor. Additional fields of view and tumor slices are shown in Supplemental Figure 1. B. Biodistribution data showing the uptake of 89Zr-scFv in an orthotopic hepatocellular tumor established from a mouse cell line derived from the Alb Cre; MYCTg; KRASG12D GEM model.

Article Snippet: Kinetic constants for the minibody and scFv antibody against human and mouse PD-L1 (Sino Biological Inc.) were determined via biolayer interferometry with an Octet RED384 instrument (ForteBio) using a previously described approach[ 11 ].

Techniques: Immunohistochemistry, Staining, Binding Assay, Expressing, Derivative Assay