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: Journal for Immunotherapy of Cancer

Article Title: Nectin4 is a novel TIGIT ligand which combines checkpoint inhibition and tumor specificity

doi: 10.1136/jitc-2019-000266

Figure Lengend Snippet: TIGIT but not DNAM1, CD112R or CD96—interacts with Nectin4. (A) FACS staining of IL-2 activated primary NK cells with Nectin4-Ig. Gray filled histogram represents background staining with secondary antibody only; black line histogram represents specific binding as indicated. (B–F) FACS staining of Raji cells transfected either with an empty vector as control (gray histograms) or with Nectin4 (black histograms). Cells were stained with commercial anti-Nectin4 mAb (B), TIGIT-Ig (C), DNAM1-Ig (D), CD112-Ig (E) or CD96-Ig (F). Figures show one representative experiment out of three performed. Graph depicting the mean fluorescence intensity values of the stainings appears in . (G) Direct binding of Nectin4 to TIGIT. The binding of fluorophore-labeled TIGIT-Ig and its ligands PVR-Ig (red) and Nectin4-Ig (green) was determined using MST. Measurements were repeated with at least three independent protein preparations. FACS, fluorescence-activated cell sorting; IL-2, interleukin-2; MST, microscale thermophoresis; NK, natural killer.

Article Snippet: Extracellular fluorescence-activated cell sorting (FACS) staining for Nectin4 (MAB2659, R&D Systems), Nectin2 (BLG-337402, BioLegend), PVR (MAB25301, R&D Systems) DNAM1 (BLG-338302, BioLegend) and TIGIT (MAB7898, R&D Systems) was performed with 0.2 μg antibody per 100 000 cells.

Techniques: Staining, Binding Assay, Transfection, Plasmid Preparation, Control, Fluorescence, Labeling, FACS, Microscale Thermophoresis

Journal: Journal for Immunotherapy of Cancer

Article Title: Nectin4 is a novel TIGIT ligand which combines checkpoint inhibition and tumor specificity

doi: 10.1136/jitc-2019-000266

Figure Lengend Snippet: Nectin4 inhibits NK cytotoxicity via TIGIT. (A, C) IL2 secretion by parental BW (A, C), (A) BW-TIGIT and (C) BW-DNAM1 cells. IL2 secretion was determined by ELISA (od 650 nm) following incubation with control anti-TIGIT or anti-DNAM1 antibodies (left in a and C) or with PVR expressing cells (right in a and C). (B, D) IL2 secretion of parental BW (B, D), (B) BW-TIGIT and (D) BW-DNAM1 cells. IL2 secretion was determined by ELISA (od 650 nm) following incubation with Raji cells transfected either with an empty vector (Raji E) as a control, or with Nectin4 (Raji N4). Figure shows one representative experiment out of 3 performed. *P<0.05. (E) FACS staining of Raji cells overexpressing Nectin4 with TIGIT-Ig. TIGIT-Ig was preincubated with no antibody (left), with a control mAb (anti-CD99 mAb clone 12E7, middle) or with anti-TIGIT blocking antibody (mAb #4 generated as described previously, right). Black line histograms represent TIGIT-Ig binding. Gray filled histograms represent background staining of the secondary antibody only. (F) Mean fluorescence intensity (MFI) values of the TIGIT-Ig staining shown in (E) relative to NO antibody staining, *p<2×10 -4 . (G) [ 35 S] methionine-labeled Raji cells transfected either with an empty vector as control (Raji empty—gray) or with Nectin4 (Raji Nectin4—black), were incubated for 5 hours with NK cells. NK cells were preincubated with no antibody (left), with a control antibody (anti-CD99 mAb clone 12E7, middle) or with an anti-TIGIT antibody (mAb #4 generated as described previously, right). The effector to target (E:T) ratios are indicated on the x-axis. Figure shows one representative experiment out of three performed. Shown is the relative average killing ±SD, *p<0.05. FACS, fluorescence-activated cell sorting; IL2, interleukin-2; NK, natural killer; NS, not significant.

Article Snippet: Extracellular fluorescence-activated cell sorting (FACS) staining for Nectin4 (MAB2659, R&D Systems), Nectin2 (BLG-337402, BioLegend), PVR (MAB25301, R&D Systems) DNAM1 (BLG-338302, BioLegend) and TIGIT (MAB7898, R&D Systems) was performed with 0.2 μg antibody per 100 000 cells.

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Control, Expressing, Transfection, Plasmid Preparation, Staining, Blocking Assay, Generated, Binding Assay, Fluorescence, Labeling, FACS

Journal: mBio

Article Title: Early enterovirus translation deficits extend viral RNA replication and elicit sustained MDA5-directed innate signaling

doi: 10.1128/mbio.01915-23

Figure Lengend Snippet: Short-term time course of PVSRIPO and CAV21 infection in a panel of in vitro tumor models (multiplicity of infection [MOI], 10). (A–D) HeLa R19 (A), A375 (B), Hs683 (C), and M059J (D) cells were infected with PVSRIPO or CAV21. Lysates collected at the indicated hpi were analyzed by immunoblot for viral translation (2C) and 2A pro -directed cleavages (eIF4G, YTHDF3; asterisks indicate eIF4G/YTHDF3 cleavage products). All assays were performed at least three times, and representative immunoblots are shown. The panels in the right column depict quantification of protein levels (% max. detected, normalized to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) loading control; means ± standard error of the mean), representing the average values from three independent series. (E) Expression of poliovirus receptor PVR (CD155) and CAV21 receptor ICAM1 (CD54) in the panel. (F) P-STAT1(Y701) response to transfection of increasing concentrations of poly(I:C) in the cell line panel. Cell lysates were collected at the indicated intervals and tested for p-STAT1(Y701)/STAT1 by immunoblot.

Article Snippet: Primary antibodies used were against eIF4G1 (#2469), GAPDH (#2118), IRF3 (#119040), p-IRF3(S386) (#37829), STAT1 (#9172), p-STAT1(Y701) (#9167), IRF7 (#49200), p-IRF7(S477) (#12390), CD54 (#67836), CD155 (#13544), IFN-β (#736710), dsRNA (K1, #28764), MDA5 (#5321), IFIT1 (#14769), and NUP98 (#2598) (all Cell Signaling Tech); and puromycin (#MABE343) (Millipore); IFN-α (#18013-1-AP), and YTHDF3 (#25537–1-AP) (both Proteintech).

Techniques: Infection, In Vitro, Western Blot, Control, Expressing, Transfection

Journal: mBio

Article Title: Early enterovirus translation deficits extend viral RNA replication and elicit sustained MDA5-directed innate signaling

doi: 10.1128/mbio.01915-23

Figure Lengend Snippet: EVs employ distinct translation sites in infected host cells (adapted from reference ). ( A ) Upon virion:CD155 binding, incoming vRNA is released into the cytoplasm from endocytic vesicles/plasma membrane invaginations. Incoming vRNA is translated upon binding of eIF4G to the viral IRES ( , ); 2A pro is released auto-catalytically from the nascent polyprotein. Rupintrivir blocks viral polyprotein processing. ( B ) PV co-opts PI4KIIIβ for generating endoplasmic reticulum (ER)-derived membrane scaffolds enriched in PI4-phosphate (PI4P) lipids, providing tethers for vRNA replication sites . ER-bound, PI4P-enriched viral replication sites form the platform for vRNA replication. PI4KIIIβ-IN-10 represses PI4P synthesis. Processive vRNA replication requires nucleoside import via the equilibrative nucleoside transporters 1 and 2 (ENT1/ENT2), which are inhibited by dipyridamole.

Article Snippet: Primary antibodies used were against eIF4G1 (#2469), GAPDH (#2118), IRF3 (#119040), p-IRF3(S386) (#37829), STAT1 (#9172), p-STAT1(Y701) (#9167), IRF7 (#49200), p-IRF7(S477) (#12390), CD54 (#67836), CD155 (#13544), IFN-β (#736710), dsRNA (K1, #28764), MDA5 (#5321), IFIT1 (#14769), and NUP98 (#2598) (all Cell Signaling Tech); and puromycin (#MABE343) (Millipore); IFN-α (#18013-1-AP), and YTHDF3 (#25537–1-AP) (both Proteintech).

Techniques: Infection, Binding Assay, Clinical Proteomics, Membrane, Derivative Assay

Journal: Clinical and experimental immunology

Article Title: CD155 expression impairs anti-PD1 therapy response in non-small cell lung cancer.

doi: 10.1093/cei/uxac020

Figure Lengend Snippet: Figure 1: (A) Representative immunohistochemistry images of CD155, PD-L1, and TIGIT from NSCLC tumor specimens (magnification, ×400). (B) The correlation between CD155 immunostaining score and PD-L1 expression. (C) Association of tumor CD155− vs CD155+ with PFS evaluated using the Kaplan–Meier method in NSCLC patients treated with αPD1 (n = 42, CD155−; n = 39, CD155+; P = 0.001). (D) Histograms of the RECIST response categories (partial response [PR], stable disease [SD], and progressive disease [PD]) from advanced NSCLC patients (n = 81) treated with αPD1 therapy. Chi-square tests by PR vs SD+PD and CD155− vs CD155+. (E) The histograms of patients with progression free response to therapy > 6 months. Chi-square tests by response > 6 months vs response < 6 months and CD155− vs CD155+. (F) Association of TIGIT− vs TIGIT+ with PFS in patients with NSCLC treated with αPD1 therapy (n = 48, TIGIT−; n = 33, TIGIT+; P = 0.092). (G) Histograms of RECIST response categories (PR, SD, and PD) from advanced patients with NSCLC (n = 81) treated with αPD1 therapy. Chi-square tests by PR vs SD+PD and TIGIT− vs TIGIT+. (H) The histograms of patients with progression-free response to therapy > 6 months. Chi-square tests by response > 6 months vs response < 6 months and CD155− vs CD155+. **P < 0.01; ns, not significant.

Article Snippet: Slices were then incubated overnight with the primary anti-CD155 antibody (81254S, Cell Signaling Technology) at 1:100 and anti-TIGIT antibody (ab243903, Abcam) at 1:100.

Techniques: Immunohistochemistry, Immunostaining, Expressing

Journal: Clinical and experimental immunology

Article Title: CD155 expression impairs anti-PD1 therapy response in non-small cell lung cancer.

doi: 10.1093/cei/uxac020

Figure Lengend Snippet: Figure 2: (A) The PFS of patients with NSCLC categorized by PD-L1 status and CD155 tumor expression. Association between PD-L1+/CD155− vs other groups evaluated using a Kaplan–Meier method. Patients were treated with αPD1 therapy (n = 30, PD-L1+/CD155+; n=28, PD-L1+/CD155−; n=9, PD-L1−/ CD155+; n=14, PD-L1−/CD155−; P=0.003). (B) Histograms of the RECIST response categories (PR, SD, PD) in NSCLC by PD-L1 status and CD155 tumor expression from advanced NSCLC patients treated with αPD1 therapy (n = 81). Chi-square tests by PR vs SD+PD and PD-L1+/CD155− vs other groups. (C) The histograms of patients with response >6 months by PD-L1 status and CD155 tumor expression. Chi-square tests by response >6 months vs response <6 months and PD-L1+/CD155− vs other groups. (D) Association of CD155− vs CD155+ with PFS evaluated using the Kaplan–Meier method in PD-L1low patients treated with αPD1 therapy (n = 8, CD155−; n = 10, CD155+; P = 0.008). (E) Histograms of CD155 status by RECIST categories (PR, SD, PD) from PD-L1low patients. Chi-square tests by PR vs SD+PD and CD155− vs CD155+. (F) The histograms of PD-L1low patients with progression-free response to therapy >6 months. Chi-square tests by response >6 months vs response <6 months and CD155− vs CD155+. (G) Association of CD155− vs CD155+ with PFS in PD-L1high patients treated with αPD1 therapy (n = 20, CD155−; n = 20, CD155+; P < 0.001). (H) Histograms of CD155 status by RECIST categories (PR, SD, PD) from PD-L1high patients. Chi-square tests by PR vs SD+PD and CD155− vs CD155+. (I) The histograms of PD-L1high patients with progression-free response to therapy >6 months. Chi-square tests by response >6 months vs response <6 months and CD155− vs CD155+. *P < 0.05; ***P < 0.001; ns, not significant.

Article Snippet: Slices were then incubated overnight with the primary anti-CD155 antibody (81254S, Cell Signaling Technology) at 1:100 and anti-TIGIT antibody (ab243903, Abcam) at 1:100.

Techniques: Expressing

Journal: Clinical and experimental immunology

Article Title: CD155 expression impairs anti-PD1 therapy response in non-small cell lung cancer.

doi: 10.1093/cei/uxac020

Figure Lengend Snippet: Figure 3: (A) Association of CD155− vs CD155+ with PFS evaluated using the Kaplan–Meier method. LUAC patients (n = 27, CD155−; n = 22, CD155+; P = 0.08) or LUSC patients (n = 15, CD155−; n = 17, CD155+; P = 0.002). (B) Histograms for CD155 status by RECIST categories (PR, SD, PD) from advanced LUAC or LUSC. Chi-square tests by PR vs SD+PD and CD155− vs CD155+. (C) The histograms of patients with progression free response to therapy > 6 months. Chi-square tests by response >6 months vs response <6 months and CD155− vs CD155+. (D) Association of CD155− vs CD155+ with PFS in PD-L1+ LUAC (n = 19, CD155−; n = 17, CD155+; P = 0.016) or LUSC (n = 9, CD155−; n = 13, CD155+; P < 0.001). (E) Histograms for CD155 status by RECIST categories (PR, SD, PD) from PD-L1+ LUAC or LUSC. Chi-square tests by PR vs SD+PD and CD155− vs CD155+. (F) The histograms of PD-L1+ patients with progression-free response to therapy > 6 months. Chi-square tests by response >6 months vs response <6 months and CD155− vs CD155+. *P < 0.05; ns, not significant.

Article Snippet: Slices were then incubated overnight with the primary anti-CD155 antibody (81254S, Cell Signaling Technology) at 1:100 and anti-TIGIT antibody (ab243903, Abcam) at 1:100.

Techniques:

Journal: Clinical and experimental immunology

Article Title: CD155 expression impairs anti-PD1 therapy response in non-small cell lung cancer.

doi: 10.1093/cei/uxac020

Figure Lengend Snippet: Figure 4: (A) Association of CD155− vs CD155+ with PFS evaluated using the Kaplan–Meier method. Patients treated with first-line αPD1 therapy (n = 18, CD155−; n = 19, CD155+; P = 0.003) or posterior-line αPD1 therapy (n = 24, CD155−; n = 20, CD155+; P = 0.025). (B) Histograms for CD155 status by RECIST categories (PR, SD, PD) from advanced patients with NSCLC who received first-line αPD1 therapy (n = 37) or posterior-line αPD1 therapy (n = 44). Chi-square tests by PR vs SD+PD and CD155− vs CD155+. (C) The histograms of patients with progression free response to therapy > 6 months. Chi-square tests by response >6 months vs response <6 months and CD155− vs CD155+. (D) Association of CD155− vs CD155+ with PFS in PD-L1+ NSCLC patients treated with first-line αPD1 therapy (n = 14, CD155−; n = 15, CD155+; P = 0.002) or posterior-line αPD1 therapy (n = 14, CD155−; n = 15, CD155+; P < 0.001). (E) Histograms for CD155 status by RECIST category (PR, SD, PD) from PD-L1+ NSCLC patients treated with first-line αPD1 therapy or posterior-line αPD1 therapy. Chi-square tests by PR vs SD+PD and CD155− vs CD155+. (F) The histograms of PD-L1+ patients with progression free response to therapy >6 months. Chi-square tests by response >6 months vs response <6 months and CD155− vs CD155+. *P < 0.05; **P < 0.01; ***P < 0.001; ns, not significant.

Article Snippet: Slices were then incubated overnight with the primary anti-CD155 antibody (81254S, Cell Signaling Technology) at 1:100 and anti-TIGIT antibody (ab243903, Abcam) at 1:100.

Techniques:

Journal: Clinical and experimental immunology

Article Title: CD155 expression impairs anti-PD1 therapy response in non-small cell lung cancer.

doi: 10.1093/cei/uxac020

Figure Lengend Snippet: Figure 5: (A) Association of tumor CD155− vs CD155+ with PFS evaluated using the Kaplan–Meier method in NSCLC patients treated with αPD1 monotherapy (n = 17, CD155−; n = 13, CD155+; P = 0.704), αPD1-combitherapy (n = 25, CD155−; n = 26, CD155+; P < 0.001). (B) Histograms for CD155 status by RECIST category (PR, SD, PD) from advanced patients NSCLC treated with αPD1-monotherapy(n = 30) or αPD1-combitherapy (n = 51). Chi-square tests by PR vs SD+PD and CD155− vs CD155+. (C) The histograms of patients with progression free response to therapy > 6 months. Chi- square tests by response>6 months vs response<6 months and CD155− vs CD155+. (D) PFS of NSCLC patients with PD-L1+ and categorized by CD155 tumor expression. Association between PD-L1+/CD155− vs PD-L1+/CD155+ evaluated using Kaplan–Meier method. Patients were treated with αPD1- monotherapy (n = 8, PD-L1+/CD155−; n = 9, PD-L1+/CD155+; P = 0.123), αPD1-combitherapy (n = 20, PD-L1+/CD155; n = 21, PD-L1+/CD155+; P < 0.001). (E) Histograms for CD155 status by RECIST categories (PR, SD, PD) from patients with advanced NSCLC treated with αPD1-monotherapy (n = 17) or αPD1-combitherapy (n = 41). Chi-square tests by PR vs SD+PD and PD-L1+/CD155− vs PD-L1+/CD155+. (F) The histograms of patients with response >6 months in PD-L1+/CD155− and PD-L1+/CD155+ group. Chi-square tests by response >6 months vs response< 6 months and PD-L1+/CD155− vs PD-L1+/ CD155+. **P < 0.01; ***P <0.001; ns, not significant.

Article Snippet: Slices were then incubated overnight with the primary anti-CD155 antibody (81254S, Cell Signaling Technology) at 1:100 and anti-TIGIT antibody (ab243903, Abcam) at 1:100.

Techniques: Expressing

Journal: Science Advances

Article Title: Human stem cell-derived β cells expressing an optimized CD155 reduce cytotoxic immune cell function for application in type 1 diabetes

doi: 10.1126/sciadv.adx9755

Figure Lengend Snippet: ( A ) Schematic of parental Mel1 INS-GFP (blue) and engineered WT (red) or Mut CD155 (green) hPSC lines [Created in BioRender. Russ, H. (2025); https://BioRender.com/r06dcp3 ]. ( B ) Immunofluorescence staining for OCT3/4 (green), NANOG (red), and 4′,6-diamidino-2-phenylindole (white) in hPSC. Scale bar represents 100 µm. ( C ) Flow cytometry for expression of CD155 in hPSC. ( D to F ) hPSC were treated with fluorescently labeled TIGIT-Ig to compare binding efficiency between the parental Mel1 (blue), CD155 WT–expressing (red), and CD155 Mut–expressing (green) lines. (D) Representative histograms of cell-bound Ig after treatment at the 25,000 ng/ml condition. (E) Differences in Ig binding efficiency characterized across a range of 0 to 25,000 ng/ml, with additional comparisons made using (F) TIGIT-Ig selectivity area under the curve (AUC) values for each binding curve, where the bound TIGIT-Ig gMFI value for each line was normalized to the Mel1-bound TIGIT-Ig gMFI at each concentration. Data reflect biological n = 3 per condition for TIGIT-Ig sBC. Significant P values are reported for one-way analysis of variance (ANOVA) with Bonferroni’s multiple comparisons of AUC values between paired samples. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: CD155 plasmid template was purchased from OriGene (RC202254), followed by transformation, amplification, and purification in New England Biolabs (NEB) 5-alpha competent Escherichia coli.

Techniques: Immunofluorescence, Staining, Flow Cytometry, Expressing, Labeling, Binding Assay, Concentration Assay

Journal: Science Advances

Article Title: Human stem cell-derived β cells expressing an optimized CD155 reduce cytotoxic immune cell function for application in type 1 diabetes

doi: 10.1126/sciadv.adx9755

Figure Lengend Snippet: ( A ) Schematic of differentiation from hPSC to sBC [Created in BioRender. Russ, H. (2025); https://BioRender.com/0d823q8 ]. ( B ) Representative bright field and pINS.GFP reporter live images of differentiating clusters at subsequent time points. ×4 or ×10 magnification images as denoted. ( C to H ) Flow cytometry analysis at day 23 of n = 6 independent differentiations of Mel1, CD155 WT, and CD155 Mutant sBC for frequency of (C) C-peptide + cells, (D) C-peptide + NKX6.1 + double-positive cells, (E) glucagon + cells, (F) total CD155 + cells or (G) CD155 + C-peptide + populations, and (H) geometric mean fluorescence intensity (gMFI) of CD155 within C-peptide + cells. ( I ) Immunofluorescence staining of day 23 sBC for insulin (green), NKX6.1 (magenta), and DAPI (white). Scale bar represents 200 µm. ( J ) Dynamic glucose stimulated insulin secretion data from perfusion study as percent of total insulin content. n = 3 independent differentiations. Significant P-values are reported for one-way ANOVA with Bonferroni’s multiple comparisons. **** P < 0.0001. ns, not significant; IBMX, 3-isobutyl-1-methylxanthine (IBMX); PP, pancreatic progenitors; DE, definitive endoderm.

Article Snippet: CD155 plasmid template was purchased from OriGene (RC202254), followed by transformation, amplification, and purification in New England Biolabs (NEB) 5-alpha competent Escherichia coli.

Techniques: Flow Cytometry, Mutagenesis, Fluorescence, Immunofluorescence, Staining

Journal: Science Advances

Article Title: Human stem cell-derived β cells expressing an optimized CD155 reduce cytotoxic immune cell function for application in type 1 diabetes

doi: 10.1126/sciadv.adx9755

Figure Lengend Snippet: sBC were treated with fluorescently labeled ( A to C ) TIGIT-Ig or ( D to F ) CD226-Ig to compare binding efficiency between the parental Mel1 (blue), CD155 WT–expressing (red), and CD155 Mut–expressing (green) lines. [(A) and (D)] Representative histograms show cell-bound Ig after treatment at the 25,000 ng/ml condition. [(B) and (E)] Differences in Ig binding efficiency were characterized across a range of 0 to 25,000 ng/ml, with additional comparisons made using [(C) and (F)] AUC values for each binding curve. Data reflect biological n = 3 per condition for TIGIT-Ig sBC and n = 4 per condition for CD226-Ig. Significant P values are reported for one-way ANOVA with Bonferroni’s multiple comparisons of AUC values between paired samples. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: CD155 plasmid template was purchased from OriGene (RC202254), followed by transformation, amplification, and purification in New England Biolabs (NEB) 5-alpha competent Escherichia coli.

Techniques: Labeling, Binding Assay, Expressing

Journal: Science Advances

Article Title: Human stem cell-derived β cells expressing an optimized CD155 reduce cytotoxic immune cell function for application in type 1 diabetes

doi: 10.1126/sciadv.adx9755

Figure Lengend Snippet: The expression of ( A and B ) HLA-A2; ( C and D ) HLA-A,B,C; ( E and F ) PD-L1; ( G and H ) CD112; and ( I and J ) CD155 was evaluated on INS-GFP + sBC following 48 hours with (orange) or without (blue) IFN-γ treatment. [(A), (C), (E), (G), and (I)] Representative histograms show staining of Mel1, CD155-WT, and CD155-Mut sBC lines relative to fluorescence-minus one (FMO) controls (black). [(B), (D), (F), (H), and (J)] Paired dot plots show gMFI values by sBC line and treatment condition. Data reflect biological n = 6 per condition. Significant P values are reported for two-way ANOVA with Bonferroni’s multiple comparisons between paired samples. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: CD155 plasmid template was purchased from OriGene (RC202254), followed by transformation, amplification, and purification in New England Biolabs (NEB) 5-alpha competent Escherichia coli.

Techniques: Expressing, Staining, Fluorescence

Journal: Science Advances

Article Title: Human stem cell-derived β cells expressing an optimized CD155 reduce cytotoxic immune cell function for application in type 1 diabetes

doi: 10.1126/sciadv.adx9755

Figure Lengend Snippet: ( A ) Experimental scheme depicting activation assay to assess the immunogenicity of Mel1 (blue), CD155 WT–expressing (red), and CD155 Mut–expressing (green) sBC by MART- or PPI-reactive avatars, as measured by flow immunophenotyping of T cells [Created in BioRender. Brown, M. (2025); https://BioRender.com/sdjn41g ]. Plots show differences in expression of the T cell activation markers, ( B to E ) CD96, ( F to I ) CD226, and ( J to M ) TIGIT on [(B) and (C), (F) and (G), and (J) and (K)] MART-1– or [(D) and (E), (H) and (I), and (L) and (M)] PPI-reactive avatars between sBC lines at the 10:1 E:T ratio compared to no sBC (purple) and no dye (black) controls. Data reflect biological n = 8 per condition. Significant P values are reported for one-way ANOVA with Bonferroni correction for multiple comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: CD155 plasmid template was purchased from OriGene (RC202254), followed by transformation, amplification, and purification in New England Biolabs (NEB) 5-alpha competent Escherichia coli.

Techniques: Activation Assay, Immunopeptidomics, Expressing

Journal: Science Advances

Article Title: Human stem cell-derived β cells expressing an optimized CD155 reduce cytotoxic immune cell function for application in type 1 diabetes

doi: 10.1126/sciadv.adx9755

Figure Lengend Snippet: ( A ) Experimental scheme depicting proliferation assay used to characterize the immunogenicity of sBC lines when cultured with allogenic naïve CD8 + T cells [Created in BioRender. Brown, M. (2025); https://BioRender.com/5fgeu0g ]. ( B ) Representative dye dilution plots depict the proliferation of naïve CD8 + T cells following 6 days of coculture with Mel1 (blue), CD155 WT–expressing (red), and CD155 Mut–expressing (green) sBC compared to no sBC (purple) and no dye (black) controls, with ( C ) violin plots showing proliferation indices for each condition (biological n = 4 per condition). Significant P values are reported for paired samples using one-way ANOVA with Bonferroni correction for multiple comparisons. *** P < 0.001 and **** P < 0.0001.

Article Snippet: CD155 plasmid template was purchased from OriGene (RC202254), followed by transformation, amplification, and purification in New England Biolabs (NEB) 5-alpha competent Escherichia coli.

Techniques: Proliferation Assay, Immunopeptidomics, Cell Culture, Expressing

Journal: Science Advances

Article Title: Human stem cell-derived β cells expressing an optimized CD155 reduce cytotoxic immune cell function for application in type 1 diabetes

doi: 10.1126/sciadv.adx9755

Figure Lengend Snippet: ( A ) Experimental scheme depicting CML assay to assess the immunogenicity of Mel1 (blue), CD155 WT–expressing (red), and CD155 Mut–expressing (green) sBC by MART- or PPI-reactive avatars, as measured by 51 Cr release, with T cell cytokine production assessed by LEGENDplex [Created in BioRender. Brown, M. (2025); https://BioRender.com/nehgyik ]. Plots show differences in cell culture supernatant concentrations following coculture with ( B to E ) MART-1– or ( F to I ) PPI-reactive avatars of [(B) and (F)] FasL, [(C) and (G)] granzyme B, [(D) and (H)] perforin, and [(E) and (I)] granulysin between sBC lines at each E:T ratio. Data reflect biological n = 8 per condition. Significant P values are reported for two-way ANOVA with Bonferroni correction for multiple comparisons between CD155 WT and CD155 Mut conditions. ( J and K ) Violin plots show percent-specific lysis of sBC by (J) MART-1– or (K) PPI-reactive avatars at each E:T ratio. Data reflect biological n = 5 per condition. Significant P values are reported for two-way ANOVA with Bonferroni correction for multiple comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: CD155 plasmid template was purchased from OriGene (RC202254), followed by transformation, amplification, and purification in New England Biolabs (NEB) 5-alpha competent Escherichia coli.

Techniques: Immunopeptidomics, Expressing, Cell Culture, Lysis

Journal: Science Advances

Article Title: Human stem cell-derived β cells expressing an optimized CD155 reduce cytotoxic immune cell function for application in type 1 diabetes

doi: 10.1126/sciadv.adx9755

Figure Lengend Snippet: ( A ) Experimental scheme depicting CML assay to assess the effect of TIGIT blockade (pink), relative to an isotype control (black) on CML of parental Mel1 and CD155 Mut–expressing sBC lines by PPI-reactive avatars, as measured by 51 Cr release, with T cell cytokine production assessed by LEGENDplex [Created in BioRender. Brown, M. (2025); https://BioRender.com/3l30agq ]. Plots show differences in cell culture supernatant concentrations following coculture with Mel1 or CD155 Mut sBC of ( B ) FasL, ( C ) granzyme B, ( D ) perforin, and ( E ) granulysin between anti (α)–TIGIT and isotype-treated PPI-reactive avatars at each E:T ratio. Data reflect biological n = 4 per condition. ( F ) Violin plots show percent-specific lysis of Mel1 (dark gray and dark pink) or CD155 Mut (light gray and light pink) sBC at each E:T ratio with or without anti-TIGIT blockade. Data reflect biological n = 4 per condition. Significant P values are reported for three-way ANOVA with Bonferroni correction for multiple comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: CD155 plasmid template was purchased from OriGene (RC202254), followed by transformation, amplification, and purification in New England Biolabs (NEB) 5-alpha competent Escherichia coli.

Techniques: Control, Expressing, Cell Culture, Lysis

Journal: Science Advances

Article Title: Human stem cell-derived β cells expressing an optimized CD155 reduce cytotoxic immune cell function for application in type 1 diabetes

doi: 10.1126/sciadv.adx9755

Figure Lengend Snippet: ( A ) Experimental scheme depicting CML assay to assess the effect of TIGIT blockade (pink), relative to an isotype control (black) on CML of Mel1 (blue), CD155 WT–expressing (red), and CD155 Mut–expressing (green) sBC by NK cells, as measured by flow immunophenotyping of NK cells and 51 Cr release assays [Created in BioRender. Brown, M. (2025); https://BioRender.com/2cr4baw ]. ( B to I ) Violin plots show differences in expression of the NK cell activation markers [(B) and (C)] CD27, [(D) and (E)] CD69, [(F) and (G)] NKG2D, and [(H) and (I)] HLA-DR between sBC lines at the 1:1 E:T ratio compared to no sBC (purple) and no dye (black) controls. Data reflect biological n = 8 per condition. Significant P values are reported for one-way ANOVA with Bonferroni correction for multiple comparisons. ( J and K ) Violin plots show percent-specific lysis of (J) Mel1, CD155 WT, and CD155 Mut sBC at each E:T ratio in isotype control cultures and (K) Mel1 (dark gray and dark pink) or CD155 Mut (light gray and light pink) sBC at each E:T ratio with or without anti-TIGIT blockade. Data reflect biological n = 6 per condition. Significant P values are reported for three-way ANOVA with Bonferroni correction for multiple comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. mAb, monoclonal antibody.

Article Snippet: CD155 plasmid template was purchased from OriGene (RC202254), followed by transformation, amplification, and purification in New England Biolabs (NEB) 5-alpha competent Escherichia coli.

Techniques: Control, Expressing, Activation Assay, Lysis