Figure 6. Down-regulation of CCR5 in systemic lymphoid tissues and inhibition of HIV-1 replication ex vivo by Dual sh1005/sh516.

Tissues were isolated from a Dual sh1005/sh516-transduced HSPC-transplanted mouse at eleven weeks post-transplantation. A. Representative data showing CCR5 expression in EGFP- and mCherry- marked human CD4⁺CD3⁺CD45⁺CD19⁻ T-cells within a gated lymphocyte population. B. Splenocytes were depleted of human CD8⁺ cells and murine CD45⁺ cells and then stimulated with PHA/IL-2 for two days. Five days later, EGFP⁺ and mCherry⁺ cells were then sorted by FACS at >97% purities. Sorted cells (5×10⁴) were then infected with either HIV-1_NL4-3 at MOI 0.5, HIV-1_{NFNSX SL9} at MOI 5.0, or HIV-1_JR-CSF at MOI 1.0 for four hours. Cells were then washed five times before culturing. HIV-1 replication was monitored by p24 ELISA analysis of culture supernatants at four and seven days post-infection. Error bars: mean + SD. Representative data of three independent experiments.