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: Neurology® Neuroimmunology & Neuroinflammation

Article Title: Differential expression of the T-cell inhibitor TIGIT in glioblastoma and MS

doi: 10.1212/NXI.0000000000000712

Figure Lengend Snippet: (A) TIGIT + , CD226 + , and CD155 + infiltrates in tumor tissue from patients with GBM and chronic active lesions from patients with MS. Arrows and insets indicate representative immunoreactivities in lymphocytes. (B) Area corresponding to TIGIT-stained GBM tissue in (A), immunolabeled with antibody against CD3. 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 t tests with a p value threshold of 0.05. ns = not significant. High magnification (scale bar = 40 μm). Low magnification (scale bar = 10 μm). GBM = glioblastoma multiforme.

Article Snippet: Serial sections were stained with primary antibodies against CD68 (Cell Signaling #76437, 1:500), MBP (Millipore Sigma MAB386, 1:500), CD3 (Dako A 40452, 1:200), TIGIT (Santa Cruz sc-103349, 1:800), CD226 (Novus Biologicals NBP1-85001, 1:50), CD155 (Bioss Inc. bs-2525R, 1:750), PD-1 (Acris Antibodies AP23805PU-M, 1:100), and PD-L1 (Spring Bioscience M4420, 1:25) and processed with the appropriate biotinylated secondary antibodies (Vector Laboratories, Burlingame CA) and avidin-biotin staining kit with diaminobenzidine as chromogen.

Techniques: Staining, Immunolabeling, Expressing

Journal: Neurology® Neuroimmunology & Neuroinflammation

Article Title: Differential expression of the T-cell inhibitor TIGIT in glioblastoma and MS

doi: 10.1212/NXI.0000000000000712

Figure Lengend Snippet: (A) PD-1 + and PD-L1 + infiltrates in tumor tissue from patients with GBM and chronic active lesions from patients with MS. Isotype control for PD-1 and PD-L1 antibody in perivascular infiltrates of MS tissue. Arrows and insets indicate representative immunoreactivities in lymphocytes. (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, as well as the number of CD3 + lymphocytes counted in the perivascular space and deep parenchyma per sample. Statistical significance was assessed by unpaired or paired Student t tests with a p value threshold of 0.05. ns = not significant. High magnification (scale bar = 40 μm). Low magnification (scale bar = 10 μm). GBM = glioblastoma multiforme.

Article Snippet: Serial sections were stained with primary antibodies against CD68 (Cell Signaling #76437, 1:500), MBP (Millipore Sigma MAB386, 1:500), CD3 (Dako A 40452, 1:200), TIGIT (Santa Cruz sc-103349, 1:800), CD226 (Novus Biologicals NBP1-85001, 1:50), CD155 (Bioss Inc. bs-2525R, 1:750), PD-1 (Acris Antibodies AP23805PU-M, 1:100), and PD-L1 (Spring Bioscience M4420, 1:25) and processed with the appropriate biotinylated secondary antibodies (Vector Laboratories, Burlingame CA) and avidin-biotin staining kit with diaminobenzidine as chromogen.

Techniques: Control

Journal: Neurology® Neuroimmunology & Neuroinflammation

Article Title: Differential expression of the T-cell inhibitor TIGIT in glioblastoma and MS

doi: 10.1212/NXI.0000000000000712

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 and CD226 and percent of CD226 + , CD4, and CD8 T cells coexpressing TIGIT (C). Histograms represent mean ± SEM. (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 the paired Student t test with a p value threshold of 0.05; ns = not significant. GBM = glioblastoma multiforme.

Article Snippet: Serial sections were stained with primary antibodies against CD68 (Cell Signaling #76437, 1:500), MBP (Millipore Sigma MAB386, 1:500), CD3 (Dako A 40452, 1:200), TIGIT (Santa Cruz sc-103349, 1:800), CD226 (Novus Biologicals NBP1-85001, 1:50), CD155 (Bioss Inc. bs-2525R, 1:750), PD-1 (Acris Antibodies AP23805PU-M, 1:100), and PD-L1 (Spring Bioscience M4420, 1:25) 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: Neurology® Neuroimmunology & Neuroinflammation

Article Title: Differential expression of the T-cell inhibitor TIGIT in glioblastoma and MS

doi: 10.1212/NXI.0000000000000712

Figure Lengend Snippet: Expression of TIGIT and CD226 measured by flow cytometry on circulating CD4 (A) and CD8 (B) T cells from healthy donors (HDs) and patients with GBM. 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 ± SEM. Statistical significance was assessed by the unpaired Student t test with a p value threshold of 0.05; ns = not significant. GBM = glioblastoma multiforme.

Article Snippet: Serial sections were stained with primary antibodies against CD68 (Cell Signaling #76437, 1:500), MBP (Millipore Sigma MAB386, 1:500), CD3 (Dako A 40452, 1:200), TIGIT (Santa Cruz sc-103349, 1:800), CD226 (Novus Biologicals NBP1-85001, 1:50), CD155 (Bioss Inc. bs-2525R, 1:750), PD-1 (Acris Antibodies AP23805PU-M, 1:100), and PD-L1 (Spring Bioscience M4420, 1:25) 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: Neurology® Neuroimmunology & Neuroinflammation

Article Title: Differential expression of the T-cell inhibitor TIGIT in glioblastoma and MS

doi: 10.1212/NXI.0000000000000712

Figure Lengend Snippet: Proliferation as measure by CellTrace™ dilution vs CD226 expression in TIGIT + CD4 and CD8 T cells at the end of a 5-day in vitro stimulation with αCD3+αCD28. Representative stainings from 2 patients presenting different baseline degrees of proliferation (A) and quantifications from 6 patients (B). Statistical significance was assessed by the paired Student t test with a p value threshold of 0.05. GBM = glioblastoma multiforme.

Article Snippet: Serial sections were stained with primary antibodies against CD68 (Cell Signaling #76437, 1:500), MBP (Millipore Sigma MAB386, 1:500), CD3 (Dako A 40452, 1:200), TIGIT (Santa Cruz sc-103349, 1:800), CD226 (Novus Biologicals NBP1-85001, 1:50), CD155 (Bioss Inc. bs-2525R, 1:750), PD-1 (Acris Antibodies AP23805PU-M, 1:100), and PD-L1 (Spring Bioscience M4420, 1:25) and processed with the appropriate biotinylated secondary antibodies (Vector Laboratories, Burlingame CA) and avidin-biotin staining kit with diaminobenzidine as chromogen.

Techniques: Expressing, In Vitro

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