Journal: Scientific Reports
Article Title: One-domain CD4 Fused to Human Anti-CD16 Antibody Domain Mediates Effective Killing of HIV-1-Infected Cells
Figure Lengend Snippet: Specific BiKE-mediated killing. ( a ) Killing of CHO-ZA-gp160 SC cells in presence of NK cells. Target cells were pre-labelled by PHK26 and incubated with serially diluted BiKEs or mD1.22-Fc followed by adding NK cells. The dead cells were measured by PI staining before evaluation by FACS analysis. Experiments were performed in triplicate and the error bars denote ± SD, n = 3. Represents NK cells incubated with mbk6 and CHO-ZA-gp160 SC cells (NK cells + mbk6 + CHO-ZA-gp160 SC , denoted as mbk6 in the figure); , NK cells + mbk11 + CHO-ZA-gp160 SC (denoted as mbk11); , NK cells + mD1.22-Fc + CHO-ZA-gp160 SC (denoted as mD1.22-Fc); other groups are negative controls , in which NK cells incubation with CHO-ZA-gp160 SC cells in the absence of BiKEs, or NK cells incubated gp160 negative CHO-ZA cells in the presence of BiKEs or mD1.22-Fc. , NK cells + mD1.22-Fc + CHO-ZA; , NK cells + CHO-ZA-gp160 SC ; , NK cells + mbk6 + CHO-ZA; , NK cells + mbk11 + CHO-ZA. ( b ) Killing of chronically HIV-1 infected cells (8E5) by human PBMCs mediated by BiKEs. Experiments were performed by co-incubation of PBMCs with BiKE opsonized 8E5 cells and the assay was developed by detecting LDH activity using Promega CytoTox-ONE™ Homogeneous Membrane Integrity kit. The anti-MERS-CoV mAb, IgG1 m336 was used as a negative control. Experiments were performed in duplicate and the error bars denote ± SD, n = 2.
Article Snippet: 8E5 cells (ATCC) were plated on round-bottom 96-well plates at a density of 2.5 × 10 4 cells in 25 μL RPMI1640 complete medium containing 20 IU/mL hIL-2 per well.
Techniques: Incubation, Staining, Infection, Activity Assay, Negative Control