Figure 1.

Vector Suppression and Mobilization in HIV-Infected Jurkat Cells

(A and B) Various vectors were generated and tested containing (A) the sh362 target site in the vector LTR (pMoHIV vectors) or with (B) a deletion of the sh362 target site in the LTR (pΔ362 vectors). The vectors also express either shRNA targeted to the LTR-362 locus, sh362, or to the HIV Tat-Rev transcript (shTat/Rev). (C) A workflow is presented depicting how HIV-infected Jurkat cells were treated and assessed for HIV-1 expression and vector mobilization. Jurkat cells were infected with HIV-1 NL4-3 MOI = 0.01, 48 hr later transduced with the vectors (MOI = 5.0), and then 48 hr later treated ±MPA (1 μM every 2 or 3 days) and followed for 28 days for p24 and qRT-PCR for vector and viral content. (D and E) The effects of vector treatment on HIV-1 p24 pg/mL in the presence (D) and absence (E) of MPA. SEMs are shown for the sh362-expressing vectors and control HIV-1-infected Jurkat cells not treated with vector or MPA. (F and G) The relative vector to virus ratio per cell found in the supernatants at day 28 in (F) MPA-treated and (G) untreated cells. (H) The observed repression of HIV-1 by sh362 is blocked by 5′AzaC treatment where shTat/Rev is not affected, suggesting sh362 functions in a transcriptional manner. For (D)–(H), the averages of triplicate-treated cultures are shown with the SDs. *p < 0.05 based on a 2-sided paired t test of HIV-infected versus vector-treated cultures.