Fig. 2: N6/αCD3-αCD28 enhances the T cell activation and T cell mediated lysis of SHIV- and HIV- infected cells.

A, B HIV dependent activation of T cells by N6/αCD3-αCD28. Effector PBMC cells were co-cultured with target cells that were either uninfected or HIV-infected CEM cells (indicated by ENV- and ENV+) in the presence of the trispecific antibodies and Brefeldin A for 16 h. The percentage of activated CD4⁺ and CD8⁺ T cells expressing both CD69 and CD25 (A) and intracellular IFN-γ (B) were measured using flow cytometry. The data were plotted as the mean ± SEM (n = 3 biological replicates). C, D Targeted lysis of HIV infected cell lines CEM-IIIb and ACH2 by N6/αCD3-αCD28. The HIV infected cells were co-cultured with SHIV⁺ LN CD8⁺ T cells which served as effector cells, in the presence of trispecific antibodies for 12 h. Percent lysis of the infected cells was measured using flow cytometry after staining with live/dead and apoptotic cell markers. Representative dose response data (C) and area under the curve (AUC) analysis (D) from three independent experiments. E Reduction in the levels of SHIV gag DNA in CD4⁺ T cells from SHIV-infected LN by N6/αCD3-αCD28. Sorted CD4⁺ and CD8⁺ T cells that served as target and effectors, respectively, were co-cultured with trispecific antibodies for 12 h. CD4⁺ T cells were then resorted from the co-culture for quantification of SHIV gag DNA. The data were plotted as the mean ± SEM (n = 5 biological replicates). F Reduction in the number of latently-infected CD4⁺ T cells by N6/αCD3-αCD28. Resting CD4⁺ T cells (CD4⁺CD25⁻CD69⁻) were sorted from PBMCs and infected with HIV BaL after culture with CCL19 for 3 days. These CD4⁺ T cells were then co-cultured with allogeneic CD8⁺ T cells and trispecific antibodies for 14 h. Intracellular p24 expression of CD4⁺ T cells was then measured by flow cytometry. The data were plotted as the mean ± SEM (n = 6 biological replicates). Statistical significance was measured by unpaired, two-tailed Student’s t-test with P values less than 0.05 considered significant (B, D, E, F). Source Data are provided as a Source Data file.