Fig. 6.

HA29-scDb induces dose-dependent viral suppression. (A) Representative flow plots from a suppression assay showing the dramatic decrease in viable GFP⁺ cells with increasing concentrations of HA29-scDb compared to cocultures containing isotype (H2) scDb. Activated CD4⁺ T cells from an A2-expressing healthy donor were infected with ΔEnv-NL4.3-EGFP and cocultured with autologous prestimulated CD8⁺T cells for 72 h before flow cytometric analysis (Materials and Methods). (B) Percent of live GFP⁺ cells remaining after 72 h of coculture with HA29-, HI12-, or H2-scDb and autologous CD8⁺ T cells. Results are expressed as a percent of the live GFP⁺ cells remaining in cultures without scDbs. (C) Suppression assays using cells from three A2⁺ and one A2⁻ donor. Results are expressed as percent of the live GFP⁺ cells remaining relative to cultures without scDbs. (D) Suppression of a replication-competent reservoir isolate 33A10 (56). Healthy donor CD4⁺ T cells were activated, infected with 33A10, and cocultured with autologous CD8⁺ T cells in in the presence of 0.25 nM of the HA29-, HI12-, or H2-scDb. Supernatant p24 was measured by ELISA after 1, 3, and 5 d of coculture. Reduction in p24 was normalized to the no scDb control. (E) Target cells (infected cells or infected cells pulsed with 10 μg/mL ILK peptide) were cocultured with autologous CD8⁺ T cells in the presence of the HI12- or H2-scDbs. Results are expressed as a percent of the live GFP⁺ cells remaining in cultures without scDbs. Data in represent the mean ± SD of three independent experiments analyzed by two-way ANOVA followed by Tukey’s multiple comparison test. Data in C represent the mean ± SD of four independent experiments analyzed by two-way ANOVA followed by Tukey’s multiple comparison test. Data in D and E represent the mean and range of two technical replicates and were analyzed by two-way ANOVA followed by Tukey’s multiple comparison test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.