human unc5b antibody  (Danaher Inc)

Journal: Science Advances

Article Title: The FLRT3-UNC5B checkpoint pathway inhibits T cell–based cancer immunotherapies

doi: 10.1126/sciadv.adj4698

Figure Lengend Snippet: ( A and B ) Human PBMCs from three healthy donors were activated with CD3 + CD28 Abs, and FLRT3 binding partner expression was analyzed on CD8 + T cells on days 0 (naïve), 3, and 4 (activated). (A) Representative histograms for UNC5B and LPHN1 staining on day 3, and (B) quantification of % CD8 + T cell expression. Each dot represents one donor, and error bars denote SD. ( C ) Representative dot plots of UNC5B and checkpoint receptor expression in CD8 + T cells from PBMCs activated with CD3 Abs for 10 days. ( D ) Illustration showing Jurkat-NG reporter system. ( E and F ) Jurkat-NG cells were transduced with LPHN3 or UNC5B and cocultured with EV- or FLRT3-expressing 293T-OKT3 cells for 16 hours, followed by GFP quantification by flow cytometry. Quantification of GFP MFI in Jurkat-NG cells with or without (E) UNC5B or (F) LPHN3 overexpression. ( G ) UNC5B–Jurkat-NG cells were cocultured with 293T-OKT3 cells expressing FLRT3, PD-L1, or CD80 for 16 hours, and NF-κB activity (GFP) was measured by flow cytometry. Quantification of GFP is shown. ( H and I ) UNC5B–Jurkat-NG cells were activated with (H) CD3 + CD28 Abs or (I) soluble CD3 Abs on control Fc of FLRT3 Fc–coated plates for 16 hours, and NF-κB activity (GFP) was measured by flow cytometry. Quantification of GFP levels is shown. For (E) to (I), each data point represents one technical replicate and error bars denote +SD. Data representative of at least two independent experiments. Statistical significance was determined for (E), (F), (H), and (I) by unpaired t test and for (G) by one-way ANOVA with Tukey’s post hoc test for multiple comparisons. For all the data, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Human UNC5B antibody , Abcam , ab104871.

Techniques: Binding Assay, Expressing, Staining, Transduction, Flow Cytometry, Over Expression, Activity Assay

Journal: Science Advances

Article Title: The FLRT3-UNC5B checkpoint pathway inhibits T cell–based cancer immunotherapies

doi: 10.1126/sciadv.adj4698

Figure Lengend Snippet: ( A ) Quantification of GFP–NF-κB in UNC5B-Jurkat-NG cells following 16 hours of coculture with EV or FLRT3-293T-OKT3 cells, and NP591 or isotype. Data representative of two experiments. ( B ) Four-day activated human PBMCs from three donors were cocultured with SKOV3 cells with CD3 + CD28 Abs in the presence of NP591 or isotype Ab. Three days later, IFN-γ was measured by ELISA. ( C and D ) PBMCs from three healthy donors (responders) were cocultured with donor PBMCs (stimulators) at a 1:1 ratio for 7 days in the presence of irradiated SKOV3 cells. Isotype control or NP591 was added in the cultures on days 0 and 3. On day 7, supernatants were harvested for IFN-γ (C) and TNF-α (D) analysis by ELISA. Bar graph data points in (A) to (D) represent one replicate, and error bars denote SD. ( E ) FLRT3-293T cells were admixed with human PBMCs and injected in mice intradermally. Mice were treated with NP591 anti–PD-1, NP591 and anti–PD-1 combo, or isotype control intraperitoneally starting on day 7 every 2 to 3 days × 3 weeks, then once per week. n = 12 per group. ( F ) Human PBMCs were injected intravenously followed by intradermal injection of SKOV3 cells 1 day later. Mice were treated with NP591 or isotype control intraperitoneally starting on day 7 every 2 to 3 days × 2 weeks, then once per week. N = 9 for isotype, n = 10 for NP591. Error bars denote SEM in (E) and (F). Data representative of two independent experiments. Statistical significance was determined for (A) by unpaired t test, for (B) by one-way ANOVA with Tukey’s post hoc test for multiple comparisons, for (C) and (D) by two-way ANOVA with Tukey’s post hoc test for multiple comparisons, and for (E) and (F) by paired t test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Human UNC5B antibody , Abcam , ab104871.

Techniques: Enzyme-linked Immunosorbent Assay, Irradiation, Injection

Journal: Science Advances

Article Title: The FLRT3-UNC5B checkpoint pathway inhibits T cell–based cancer immunotherapies

doi: 10.1126/sciadv.adj4698

Figure Lengend Snippet: List of commercial FACS antibodies/reagents used for cell staining.

Article Snippet: Human UNC5B antibody , Abcam , ab104871.

Techniques: Staining, Purification

Journal: Science Advances

Article Title: The FLRT3-UNC5B checkpoint pathway inhibits T cell–based cancer immunotherapies

doi: 10.1126/sciadv.adj4698

Figure Lengend Snippet: List of binding ELISA-related proteins/antibodies.

Article Snippet: Human UNC5B antibody , Abcam , ab104871.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant