Fig. 5

MU-CAR-T cells efficiently suppressed HIV-1 rebound after the withdrawal of antiviral treatment in vitro. A Flow chart of the experimental design. PHA-activated CD4⁺ T cells were infected with wild-type HIV-1_{NL4 − 3}. On Day 6, inhibitory drugs including AZT and Lopinavir were added to inhibit viral replication. On Day 14, anti-HIV-1 drugs were withdrawn, and infected CD4⁺ T cells were co-cultured with various groups of T cells including control CD8⁺ T cells, SdΔN17-28BBZ3-expressing CD8⁺ T cells, conventional CAR-T and VRC01-MU-CAR-T cells. On Day 26, infected CD4⁺ T cells were harvested to detect viral RNAs. HIV-1 p24 proteins within the supernatant were monitored every 2 days. B The expression of activation markers including CD25 and HLA-DR was analyzed on unstimulated and activated CD4⁺ T cells, as well as CD4⁺ T cells on Day 14 post infection. The percentage of cells was indicated in each quadrant. C Culture supernatants were tested for the presence of p24 utilizing ELISA assay every 2 days, which represented the expression and rebound of HIV-1. Data represented as mean ± SD. N = 3 independent biological replicates. D Cells from each group were harvested on Day 26, and cell-associated HIV-1 RNAs were quantified utilizing real-time RT-qPCR. Data represented as mean ± SD. N = 3 independent biological replicates. Data in (C) were analyzed by one-way ANOVA with Tukey’s multiple comparisons test. Data in (D) were analyzed by two-way ANOVA with Tukey’s multiple comparisons test. **** p < 0.0001