The lack of restriction in macaque DCs is the result of a dysfunctional TRIM5α protein. (A) Quantitative RT-PCR was carried out on human and macaque PBLs and DCs using specific primers to amplify either TRIM5α or CypA mRNA. Results are represented as mean relative expression ratios TRIM5α/CypA mRNA plus or minus SEM. (B) Quantitative RT-PCR was carried out on macaque PBLs and DCs to amplify TRIM5α, TRIM5γ, or CypA mRNA. Results show mean relative ratios for TRIM5α/CypA (left), TRIM5γ/CypA (middle), and TRIM5α/TRIM5γ (right) plus or minus SEM. (C) Rhesus TRIM5α was inserted downstream of the CMV promoter in a TRIP HIV-1–derived vector (TRIP ΔU3 CMV TRIM5α). Macaque DCs or P4-CCR5 indicator cells (Hela-CD4-LTR-LacZ)⁸  were transduced with TRIP ΔU3 CMV TRIM5α (rhTRIM5α) or with TRIP ΔU3 CMV GFP (mock). After 48 hours, cells were infected either with HIV-1 LAI-Luc, bearing the F-Luc gene downstream of env (DCs) or with HIV-1 NL43 (P4-CCR5). After a further 48 hours, cells were lysed and plate luminescence was used to measure luciferase and β-gal activity in DCs and P4-CCR5 cells, respectively. Results are represented as percentage (± SD) of the highest mock dose.