Journal: Frontiers in Immunology

Article Title: Redirection of Cord Blood T Cells and Natural Killer Cells for Elimination of Autologous HIV-1-Infected Target Cells Using Bispecific DART® Molecules

doi: 10.3389/fimmu.2020.00713

Figure Lengend Snippet: Redirection of cord blood mononuclear cell (CBMC)-derived cytolytic T cells for elimination of autologous HIV-1 infected CD4 T cells by HIV × CD3 DART molecules is not improved by incubation with (A) 10 ng/mL exogenous IL-12 (CBMC n = 8, PBMC n = 5), (B) costimulation with anti-CD28 or anti-CD137 antibodies (CBMC n = 3, PBMC n = 5), or (C) a combination of costimulation with anti-CD28 and anti-CD137 (CoStim) and exogenous IL-12 (CBMC n = 4, PBMC n = 3). (D) HIV × CD3 DART molecule-mediated elimination of autologous HIV-1-infected CD4 + T cells is improved when CD8 + T cells used as effectors are activated by T cell receptor stimulation (anti-CD28, anti-CD3, and IL-2) for 48 h prior to use in the cell killing assays (CBMC and PBMC n = 5). Box plots represent the interquartile ranges, horizontal lines indicate the medians, and error bars extend to the minimum and maximum observed values. 24 h killing assays with a CD8 + effector to HIV-1 4403bmC5-infected autologous CD4 + target cell ratio of 30:1. All data reported as positive area under the dilution curve (pAUC).

Article Snippet: After 48 h of infection, the CD4 + T cells were incubated with CD8 + T cells purified from autologous PBMC or CBMC using negative selection with magnetic beads (human CD8 + T cell isolation kit, Miltenyi Biotec) at a CD8 + T cell to target cell ratio of 30:1 in 1⁄2 area opaque flat bottom plates (Corning Life Sciences, Corning, NY).

Techniques: Derivative Assay, Infection, Incubation