Journal: Cancers

Article Title: Chemotherapeutics Used for High-Risk Neuroblastoma Therapy Improve the Efficacy of Anti-GD2 Antibody Dinutuximab Beta in Preclinical Spheroid Models

doi: 10.3390/cancers15030904

Figure Lengend Snippet: Impact of chemotherapy on percentage of ( A ) cytotoxic NK cells of lymphocytes and NK-cell-specific activating receptors. ( B – E ) 5 × 10 6 PBMCs were treated for 24 h under cell culture conditions with either carboplatin (2 µg/mL, open circles), cisplatin (1 µg/mL, open triangles), etoposide (0.5 µg/mL, open squares), vincristine (0.05 µg/mL, open diamonds), or the cyclophosphamide metabolite (4-HPC, 1 µg/mL, open hexagons). After 72 h of culturing, cells were analyzed for NKp30, NKp44, NKp46, NKG2D, and CD226 expression, using flow cytometry. ( A ) Relative number of cytotoxic NK cells (CD3 − , CD56 dim ) in lymphocytes. ( B – E ) Geometric mean fluorescence intensity (gMFI) of respective activating receptor of cytotoxic NK cells after chemotherapy. Data represent at least four biological replicates. Means and SEM are indicated as black lines and error bars, respectively. For statistical analysis, repeated measures ANOVA with appropriate post hoc test was used; * p < 0.05 vs., ** p < 0.01 versus untreated control (medium).

Article Snippet: Incubation with the following antibodies in a total volume of 100 μL was conducted for 20 min at RT: CD3-VioGreen (REA613, 1:200), CD56-APC-Vio770 (REA196, 1:200), CD226-VioBlue (REA1040, 1:50); CD335 (NKp46)-Vio Bright B515 (REA808, 1:50), CD337 (NKp30)-PE (REA823, 1:75), CD336 (NKp44)-APC (REA1163, 1:75), and CD314 (NKG2D)-PE-Vio 770 (REA1228, 1:75), all from Miltenyi Biotec.

Techniques: Cell Culture, Expressing, Flow Cytometry, Fluorescence, Control

Journal: bioRxiv

Article Title: Using peptide-exchange systems to interrogate peptide-specific KIR binding to HLA Class I

doi: 10.64898/2026.03.03.708729

Figure Lengend Snippet: (A) Display of ULBP1 and CD155 on CombiCells detected by mAbs and NKG2D-Fc, and DNAM-1-Fc, respectively. (B&C) KIR2DL1+ and KIR2DL1-NK-cell degranulation (CD107a upregulation) in response to ULBP1, CD155, and HLA-C*05:01 displayed on CombiCells. (C) Three types of HLA-C*05:01 were tested, all containing P2 (IIDKSGSTV); wild-type, open and dipeptide exchanged. Data from three independent experiments with NK cells from separate donors are shown.

Article Snippet: For binding assays, APC-conjugated Fc fusion protein (NKG2D-Fc (R&D Systems, Cat. No: 1299-NK) or DNAM-1-Fc (R&D Systems, Cat. No: # 666-DN) or KIR-Fc (R&D Systems, KIR2DL1-Fc (Cat. No: 1844-KR-050), KIR2DL3-Fc (Cat. No: 2014-KR-050), KIR2DS4-Fc (Cat. No: 1847-KR-050) (3.6 μg mL -1 ) was added at 25 μL per well and incubated for 30 min at 4 °C in the dark.

Techniques:

Journal: bioRxiv

Article Title: Differential Expression of the T cell Inhibitor TIGIT in Glioblastoma and Multiple Sclerosis

doi: 10.1101/591131

Figure Lengend Snippet: (A) TIGIT + , CD226 + and CD155 + infiltrates in tumor tissue from GBM patients and chronic active lesions from MS patients. (B) CD155 expression in tumor cells and cortical neurons. (C) Quantification of TIGIT + , CD226 + and CD155 + infiltrates in GBM tumor tissue and MS lesions. Statistical significance was assessed by unpaired Student’s t tests with a p-value threshold of 0.05. ns = non-significant. Scale bars = 40 μm.

Article Snippet: Serial sections were stained with primary antibodies against CD3 (Dako A 40452, 1:200), TIGIT (Santa Cruz sc-103349, 1:800), CD226 (Novus Biologicals NBP1-85001, 1:50), and CD155 (Bioss Inc. bs-2525R, 1:750), and processed with the appropriate biotinylated secondary antibodies (Vector Laboratories, Burlingame CA) and avidin-biotin staining kit with diaminobenzidine as chromogen.

Techniques: Expressing

Journal: bioRxiv

Article Title: Differential Expression of the T cell Inhibitor TIGIT in Glioblastoma and Multiple Sclerosis

doi: 10.1101/591131

Figure Lengend Snippet: (A) PD-1 + and PD-L1 + infiltrates in tumor tissue from GBM patients and chronic active lesions from MS patients. (B) Quantification of PD-1 + and PD-L1 + infiltrates in GBM tumor tissue and MS lesions. (C) Frequency of TIGIT + and CD226 + lymphocytes in the perivascular space and deep parenchyma. Statistical significance was assessed by unpaired Student’s t tests with a p-value threshold of 0.05. ns = not significant. Scale bar = 40 μm.

Article Snippet: Serial sections were stained with primary antibodies against CD3 (Dako A 40452, 1:200), TIGIT (Santa Cruz sc-103349, 1:800), CD226 (Novus Biologicals NBP1-85001, 1:50), and CD155 (Bioss Inc. bs-2525R, 1:750), and processed with the appropriate biotinylated secondary antibodies (Vector Laboratories, Burlingame CA) and avidin-biotin staining kit with diaminobenzidine as chromogen.

Techniques:

Journal: bioRxiv

Article Title: Differential Expression of the T cell Inhibitor TIGIT in Glioblastoma and Multiple Sclerosis

doi: 10.1101/591131

Figure Lengend Snippet: (A) Expression of TIGIT and CD226 measured by flow cytometry on CD4 and CD8 T cells from tumor infiltrates. (B) Percent of CD4 and CD8 T cells expressing TIGIT, CD226, and percent of CD226 + , CD4, and CD8 T cells co-expressing TIGIT (C). Histograms represent mean ± S.E.M. (D) Quantification of the frequency of TIGIT + , CD226 + , and TIGIT + among CD226 + for CD4 and CD8 circulating and tumor-infiltrating T cells. Frequencies were assessed by flow cytometry. Dots connected by a line represent the same patient. TIL = tumor infiltrating lymphocytes. Statistical significance was assessed by paired Student’s t test with a p-value threshold of 0.05; ns = non-significant.

Article Snippet: Serial sections were stained with primary antibodies against CD3 (Dako A 40452, 1:200), TIGIT (Santa Cruz sc-103349, 1:800), CD226 (Novus Biologicals NBP1-85001, 1:50), and CD155 (Bioss Inc. bs-2525R, 1:750), and processed with the appropriate biotinylated secondary antibodies (Vector Laboratories, Burlingame CA) and avidin-biotin staining kit with diaminobenzidine as chromogen.

Techniques: Expressing, Flow Cytometry

Journal: bioRxiv

Article Title: Differential Expression of the T cell Inhibitor TIGIT in Glioblastoma and Multiple Sclerosis

doi: 10.1101/591131

Figure Lengend Snippet: Expression of TIGIT and CD226 measured by flow cytometry on circulating CD4 (A) and CD8 (B) T cells from healthy donors (HD) and GBM patients. Quantification of the frequency of TIGIT + , CD226 + , and TIGIT + among CD226 + and PD-1 + for CD4 (C) and CD8 (D) circulating T cells. The values for GBM are the same as depicted in in the “Blood” group. Histograms represent mean ± S.E.M. Statistical significance was assessed by unpaired Student’s t test with a p-value threshold of 0.05; ns = non-significant.

Article Snippet: Serial sections were stained with primary antibodies against CD3 (Dako A 40452, 1:200), TIGIT (Santa Cruz sc-103349, 1:800), CD226 (Novus Biologicals NBP1-85001, 1:50), and CD155 (Bioss Inc. bs-2525R, 1:750), and processed with the appropriate biotinylated secondary antibodies (Vector Laboratories, Burlingame CA) and avidin-biotin staining kit with diaminobenzidine as chromogen.

Techniques: Expressing, Flow Cytometry