Journal: Diagnostics

Article Title: Prognostic and Predictive Significance of B7-H3 and CD155 Expression in Gastric Cancer Patients

doi: 10.3390/diagnostics15212695

Figure Lengend Snippet: Fold change expression levels of B7-H3 and CD155 across individual gastric cancer cases. A fold change greater than 1 was interpreted as positive or upregulated gene expression, where fold regulation was considered equivalent to the fold change value. A fold change greater than 2 was classified as moderate to high upregulation in gene expression.

Article Snippet: For B7-H3, a recombinant rabbit monoclonal IgG antibody (HUABIO, Woburn, MA, USA), clone: HA721245; dilution 1:5000) was applied, while for CD155, a recombinant rabbit monoclonal IgG antibody (Proteintech, Rosemont, IL, USA, clone: 241405F5; dilution 1:1500) was used.

Techniques: Expressing, Gene Expression

Journal: Diagnostics

Article Title: Prognostic and Predictive Significance of B7-H3 and CD155 Expression in Gastric Cancer Patients

doi: 10.3390/diagnostics15212695

Figure Lengend Snippet: Comparison of B7-H3 and CD155 fold-Change Expression According to Metastatic Status (M0 vs. M1). Fold change was calculated by dividing the normalized gene expression in the test sample by the normalized gene expression in the control sample. Fold regulation represents the biologically relevant direction and magnitude of change derived from the fold change value.

Article Snippet: For B7-H3, a recombinant rabbit monoclonal IgG antibody (HUABIO, Woburn, MA, USA), clone: HA721245; dilution 1:5000) was applied, while for CD155, a recombinant rabbit monoclonal IgG antibody (Proteintech, Rosemont, IL, USA, clone: 241405F5; dilution 1:1500) was used.

Techniques: Comparison, Expressing, Gene Expression, Control, Derivative Assay

Journal: PLoS ONE

Article Title: The Role of T Cell Costimulation via DNAM-1 in Kidney Transplantation

doi: 10.1371/journal.pone.0147951

Figure Lengend Snippet: Schematic illustration of the DNAM-1 pathway in the interaction between a T cell and an antigen presenting cell (APC). The T cell recognizes its cognate antigen in the context of MHC with its T cell receptor (TCR). For further activation it needs costimulatory signals, which can be delivered via DNAM-1 (CD226) binding to its two ligands CD155 and CD112 expressed on the APC. CD155 and CD112 also bind TIGIT, a co-inhibitory receptor of the Ig-family. CD155 has an additional receptor called CD96.

Article Snippet: For detection of CD155 by immunohistochemistry a rabbit polyclonal antibody against murine CD155 purchased from Biorbyt (Cambridge, UK).

Techniques: Activation Assay, Binding Assay

Journal: PLoS ONE

Article Title: The Role of T Cell Costimulation via DNAM-1 in Kidney Transplantation

doi: 10.1371/journal.pone.0147951

Figure Lengend Snippet: (A) Primary B6-derived rTECs were left untreated or stimulated with IFN-β and -γ (100 U/ml each) for 48 hours. Surface expression of CD155 and CD112 was analyzed by FACS. Shaded: isotype control, dotted line: unstimulated, solid line: stimulated. A representative result of 4 independent experiments is shown. (B-F) Renal allografts from B6 to fully MHC-mismatched CBA recipients were performed. B6 to B6 syngeneic renal grafts were performed as control. In each group we included at least 5 mice. (B) Real time-PCR for CD155 and CD112 was performed on naïve kidneys and renal syn- and allografts. The fold upregulation of CD112 and CD155 compared to naïve renal tissue is depicted. Groups were compared using the Mann-Whitney-test: * P = 0.016, ns = not significant. (C) The expression of CD155 and CD112 in this group of grafts highly correlated (P<0.001). (D-F) Immunohistochemical staining for CD155 was performed. (D) An overview shows the tubular expression of CD155 in both syn- and allografts (arrows, scale bar 200 μm). (E) The expression of CD155 is located mainly in the medulla in syngrafts and is increased in the cortical area in allografts. (F) The papillae of syngrafts show no staining for CD155, whereas in allografts tubuli in the papillae exhibit strong CD155 staining (scale bar 200 μm).

Article Snippet: For detection of CD155 by immunohistochemistry a rabbit polyclonal antibody against murine CD155 purchased from Biorbyt (Cambridge, UK).

Techniques: Derivative Assay, Expressing, Control, Real-time Polymerase Chain Reaction, MANN-WHITNEY, Immunohistochemical staining, Staining

Journal: PLoS ONE

Article Title: The Role of T Cell Costimulation via DNAM-1 in Kidney Transplantation

doi: 10.1371/journal.pone.0147951

Figure Lengend Snippet: (A-C) Isolated CBA T cells were stimulated with irradiated B6 splenocytes. CD8 or CD4 T cells were either cultured alone or in combination (ratio 1:2). An anti-DNAM-1 antibody (3B3) was added to the culture at 25 μg/ml. As control an unspecific isotype control antibody was used at the same concentration. (A) Proliferation was measured on day 4 of culture. * P = 0.035, ** P<0.01. (B) IFN-γ was measured in the supernatant from the same CD8+CD4 cultures. (C) Cytotoxicity of CBA splenocytes stimulated in the presence or absence of 3B3 (25 μg/ml) against IFN-stimulated B6 WT rTECs was measured on day 5. (D-F) Isolated B6 T cells were stimulated with irradiated BALB/c WT or CD155 KO splenocytes. CD8 or CD4 T cells were either cultured alone or in combination (ratio 1:2). (D) Proliferation was measured on day 4 of culture. ** P<0.01. (E) IFN-γ was measured in the supernatant from the same CD8+CD4 cultures. (F) Cytotoxicity of the CD8+CD4 culture against IFN-stimulated WT BALB/c rTECs was measured on day 5. (G-I) Isolated CBA T cells were stimulated with irradiated B6 WT or CD112 KO splenocytes. CD8 or CD4 T cells were either cultured alone or in combination (ratio 1:2). (G) Proliferation was measured on day 4 of culture. * P = 0.02. (H) IFN-γ was measured in the supernatant from the same CD8+CD4 cocultures. (I) Cytotoxicity of CBA splenocytes stimulated with irradiated WT B6 or CD112 KO splenocytes against IFN-stimulated WT B6 rTECs was measured on day 5. Data points represent mean values of triplicates. All experiments were performed at least 3 times. Representative figures are displayed.

Article Snippet: For detection of CD155 by immunohistochemistry a rabbit polyclonal antibody against murine CD155 purchased from Biorbyt (Cambridge, UK).

Techniques: Isolation, Irradiation, Cell Culture, Control, Concentration Assay

Journal: PLoS ONE

Article Title: The Role of T Cell Costimulation via DNAM-1 in Kidney Transplantation

doi: 10.1371/journal.pone.0147951

Figure Lengend Snippet: (A) CBA splenocytes were stimulated with irradiated BALB/c splenocytes. Cytotoxicity against IFN-stimulated WT BALB/c or CD155 KO rTEC targets was measured on day 5 of coculture. (B) CBA splenocytes were stimulated with irradiated B6 splenocytes. Cytotoxicity against IFN-stimulated WT or CD112 KO targets was measured on day 5 of coculture. (C) CBA splenocytes were stimulated with irradiated BALB/c splenocytes. Cytotoxicity against IFN-stimulated CD155 KO rTEC targets in the presence (25 μg/ml) or absence of a blocking anti-CD112 antibody was measured on day 5 of coculture. (D) CBA splenocytes were stimulated with irradiated B6 splenocytes. Cytotoxicity against IFN-stimulated WT targets was measured on d 5 of coculture in the presence or absence of a blocking anti-DNAM-1 antibody (50 μg/ml). Data points represent mean values of triplicates. All experiments were performed at least 3 times. Representative figures are displayed.

Article Snippet: For detection of CD155 by immunohistochemistry a rabbit polyclonal antibody against murine CD155 purchased from Biorbyt (Cambridge, UK).

Techniques: Irradiation, Blocking Assay

Journal: PLoS ONE

Article Title: The Role of T Cell Costimulation via DNAM-1 in Kidney Transplantation

doi: 10.1371/journal.pone.0147951

Figure Lengend Snippet: Renal allografts were performed in a non-life supporting manner. All allografts were harvested on day 21. Strain combinations were fully MHC-mismatched: (A) BALB/c WT (n = 5) or CD155 KO (n = 5) into B6 and (B) B6 WT (n = 5) or CD112 KO (n = 3) into CBA. Representative H & E stainings are shown. All allografts displayed severe interstitial infiltrates as well as tubulitis in more than 50% of the graft. Furthermore, arteritis was detected in all grafts classifying them to Banff grade II or III. The chosen pictures are taken from allografts with the following Banff grades: (A) BALB/c to B6: IIA; CD155 KO to B6: IIB; (B) B6 to CBA: IIB; CD112 to CBA: III (scale bar 200 μm). (C) Apoptotic cells in renal allografts were detected by immunohistochemical staining for ssDNA. (D) Representative picture of a necrotic area in an H & E slide of a CD155 KO renal allograft (scale bar 1200 μm). (E) The area of necrotic tissue in H & E stained slides from renal allografts was detected by scanning them at a resolution of 0.23 μm. Quantification was performed using NDPView software. Groups were compared using the Mann-Whitney-test.

Article Snippet: For detection of CD155 by immunohistochemistry a rabbit polyclonal antibody against murine CD155 purchased from Biorbyt (Cambridge, UK).

Techniques: Immunohistochemical staining, Staining, Software, MANN-WHITNEY

Journal: Oncology Letters

Article Title: Antitumor effects of NK cells expanded by activation pre‑processing of autologous feeder cells before irradiation in colorectal cancer

doi: 10.3892/ol.2023.13818

Figure Lengend Snippet: Combination of anti-CD3 mAb, IFN-r, and IL-2 with radiation further promotes the expression levels of activating ligands in human PBMCs. PBMCs were cultured with or without anti-CD3 mAb, IFN-r, and IL-2 and then irradiated with 25 Gy. The cells were cultured for 0, 24, 48, or 72 h. CD48, CD112, CD155, and natural killer group 2D ligands (MICA, MICB, ULBP-1, ULBP-2/5/6, and ULBP-3) were analyzed by flow cytometry. (A, C, and E) Ratios of MFIs obtained from irradiated PBMCs and activated/irradiated PBMCs. Relative expression ratios were calculated by dividing the MFI of irradiated PBMCs (24, 48, and 72 h) or activated/irradiated PBMCs (24, 48, and 72 h) by that of untreated PBMCs (0 h). (B, D, and F) Representative flow cytometry histograms of each donor [white, untreated PBMCs (0 h); bright gray, irradiated PBMCs (48 h); gray, activated/irradiated PBMCs (48 h); dark gray, irradiated PBMCs (72 h); black, activated/irradiated PBMCs (72 h)]. The data are presented as the mean ± SD of 3 donors. Statistical significance was determined using paired and unpaired Student's t-test. # P<0.05, ## P<0.005, ### P<0.0005 [untreated PBMCs (0 h) vs. 24, 48, or 72 h irradiated or activated/irradiated PBMCs]. $ P<0.05, $$ P<0.005, $$$ P<0.0005 (24, 48, and 72 h irradiated PBMCs vs. 24, 48, and 72 h activated/irradiated PBMCs). mAb, monoclonal antibody; r, recombinant; PBMCs, peripheral blood mononuclear cells; MFI, median fluorescence intensity; MIC, MHC class I polypeptide-related sequence; ULBP, UL16 binding protein.

Article Snippet: The cells were irradiated with or without a dose of 25 Gy in a blood irradiator (Eckert & Ziegler), and cultured for 0, 24, 48, and 72 h. The cells were incubated with antibodies against human CD48-fluorescein isothiocyanate (FITC; 1:50; BD Pharmigen; BD Biosciences; cat. no. 555759), CD112-phycoerythrin (PE; 1:50; BD Pharmigen; BD Biosciences; cat. no. 551057), CD155-PE (1:50; R&D Systems, Inc.; cat. no. FAB25301P) and NKG2D ligands [including MHC class I polypeptide-related sequence A (MICA)-PE (1:50; R&D Systems Inc.; cat. no. FAB1300P), MHC class I polypeptide-related sequence B (MICB)-PE (1:50; R&D Systems Inc.; cat. no. FAB1599P), UL16 binding protein (ULBP)-1-PE (1:50; R&D Systems Inc.; cat. no. FAB1380P), ULBP-2/5/6-PE (1:50; R&D Systems Inc.; cat. no. FAB1298P) and ULBP-3-PE (1:50; R&D Systems Inc.; cat. no. FAB1517P)] for 20 min in the dark at room temperature.

Techniques: Expressing, Cell Culture, Irradiation, Flow Cytometry, Recombinant, Fluorescence, Sequencing, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Expression of TIGIT/CD155 and correlations with clinical pathological features in human hepatocellular carcinoma

doi: 10.3892/mmr.2019.10641

Figure Lengend Snippet: Expression of CD4, TIGIT and CD155 in HCC tissues at different pathological stages and adjacent tissues. Paracancerous tissues (G1) and HCC tissues (G2-6) were stained by immunohistochemistry (×400 magnification). (A) Staining of CD4 and (B) statistical analysis of CD4-positive dots in paracancerous and HCC tissues. (C) Staining of TIGIT and (D) statistical analysis of TIGIT-positive dots in paracancerous and HCC tissues. (E) Staining of CD155 and (F) statistical analysis of CD155-positive dots in paracancerous and HCC tissues. (G) Relative mRNA expression levels of CD4, TIGIT and CD155. All data from at least three independent experiments were measured by the band density, which were normalized to GAPDH. Bar graphs (mean ± SEM) and representative images are shown. *P<0.05. G1, paracancerous tissue; G2, highly differentiated group; G3, high and medium differentiation; G4, medium differentiation; G5, medium and low differentiation; G6, low differentiation. HCC, hepatocellular carcinoma; TIGIT, T cell immunoglobulin and ITIM domain.

Article Snippet: Rabbit CD155 monoclonal antibody (cat. no. ab103630; Abcam) and rabbit TIGIT monoclonal antibody (cat. no. ab106311; Abcam) were diluted in antibody diluent (1:500/1:1,000, respectively; Beyotime Institute of Biotechnology) and rabbit GAPDH antibody (cat. no. bs-2188R; BIOSS, Beijing, China) was diluted in antibody diluent (1:4,000).

Techniques: Expressing, Staining, Immunohistochemistry

Journal: Molecular Medicine Reports

Article Title: Expression of TIGIT/CD155 and correlations with clinical pathological features in human hepatocellular carcinoma

doi: 10.3892/mmr.2019.10641

Figure Lengend Snippet: Expression of TIGIT and CD155 in different patients with HCC. (A) Protein expression of TIGIT was detected by western blotting. (B) Protein expression of CD155 was detected by western blotting. Data are expressed as the mean ± SD (*P<0.05 vs. tumor). 1, 2, 3, 4 and 5 refer to patients 1, 2, 3,4 and 5, respectively. T, tumor tissue; A, adjacent tissue; TIGIT, T cell immunoglobulin and ITIM domain.

Article Snippet: Rabbit CD155 monoclonal antibody (cat. no. ab103630; Abcam) and rabbit TIGIT monoclonal antibody (cat. no. ab106311; Abcam) were diluted in antibody diluent (1:500/1:1,000, respectively; Beyotime Institute of Biotechnology) and rabbit GAPDH antibody (cat. no. bs-2188R; BIOSS, Beijing, China) was diluted in antibody diluent (1:4,000).

Techniques: Expressing, Western Blot