Human CD160 Antibody

Detection of CD160 in Human Blood Lymphocytes by Flow Cytometry.

Human peripheral blood lymphocytes were stained with Mouse Anti-Human CD160 Monoclonal Antibody (Catalog # MAB6700) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B) and Mouse Anti-Human CD8a PE-conjugated Monoclonal Antibody (Catalog # FAB1509P). Quadrant markers were set based on control antibody staining (Catalog # MAB0041).

Detection of Human CD160 by Functional

Triggering of CD160-GPI is consistent with a positive co-stimulation role. A) Triggering of primary CD4+ T-cells with either plate-bound anti-CD3 (1 μg/ml) and anti-CD28 (0.5 μg/ml) or anti-CD3, anti-CD28 and HVEM-Fc (0.2 μg/ml) in the presence or absence of either anti-HVEM (left panel) or anti-CD160 clone CL1-R2 (right panel). IL-2 was measured in the supernatant by ELISA at 24 h post stimulation. Iso-IgG represents the matched isotype control antibody. P values were determined by two-tailed paired t test (data from three independent healthy donors). B) Left panels: Surface expression of CD160-GPI and CD160-TM on Jurkat-NFAT-Luc cells stably-transfected with CD160 plasmids. Mock-transfected cells (light grey histograms in middle and right panels) were used to set the positive and negative gates for FACS. CD160-TM is weakly detected with CD160-GPI antibodies (BY55 clone). Right panels: Quantitative RT-PCR for CD160-GPI and CD160-TM isoforms in Jurkat cells over-expressing either CD160-GPI or CD160-TM, values are relative to the house-keeping GAPDH gene transcripts (One representative experiment, n = 2). Non-transfected Jurkat (control cells) and HeLa cells were used as additional negative controls for CD160 expression. The left graph represents results with a set of Taqman probes that were not isoform selective and hybrdize both CD160-GPI and CD160-TM to demonstrate similar RNA expression levels, whereas the right graph used a set of probes that were CD160-TM specific to confirm the exclusive expression of the different CD160 isoforms in the two cell lines. C) Simultaneous triggering of TCR and CD160 using magnetic Dynal beads coated with anti-CD3, anti-CD28 and either HVEM-Fc (left panels), CD160 monoclonal antibodies (right panels) or their matched IgGs. Cell activation was monitored by measuring the absolute luciferase counts. Control cells are original Jurkat-NFAT-Luc cells non-transfected with either of the CD160 isoforms. NS: non-stimulated. P values were calculated by non-parametric two-tail t test (Mann–Whitney). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25179432), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CD160 by Functional

Differential inhibition of HVEM/CD160 binding with benchmark tool antibodies. TRF assay measuring the potency of different antibodies to inhibit the binding of recombinant human HVEM-Fc chimera to CD160+ CHO-K1 cells. A) CD160 monoclonal antibodies inhibit binding of HVEM-Fc to both CD160-GPI and CD160-TM isoforms. B) Polyclonal HVEM (left panel) and monoclonal HVEM (right panel) antibodies both enhance binding of HVEM-Fc to CD160-TM isoform. The polyclonal anti-HVEM inhibits HVEM-Fc binding to CD160-GPI (left panel). Antibody concentrations are plotted on the X axis whereas, the calculated percentage of inhibition of binding is plotted on the Y axis. Matched isotype control antibody for each individual antibody candidate was also used in the assay (empty circles and squares). CTL = control, mAb = monoclonal antibody, pAb = polyclonal antibody. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25179432), licensed under a CC-BY license. Not internally tested by R&D Systems.