FIG. 3.

Murabutide inhibits HIV-1 gene expression and viral DNA content in endogenously infected CD8-depleted blasts. Following PHA activation, CD8-depleted blasts from four HIV-1 patients were cultured in the absence or presence of 10 μg of Murabutide/ml. (A) Total RNA was extracted after a 3-day culture period, and samples (33, 100, and 300 ng) were subjected to RT-PCR amplification with primer pair GAG06-GAG04 to detect unspliced Gag or Pol mRNA and with primer pair BSS-KPNA to detect intermediate-size singly spliced viral transcripts. These mRNAs were named on the basis of the exons they contain and the proteins they produce (41). Constitutively expressed β-actin mRNA in the same samples was also amplified. An equivalent amount of RNA from the 8E5 cell line was used as positive control for RT-PCR amplification. The HIV-1 isolates in cultures from patients 1 and 2 were X4 and R5 tropic, respectively. (B) Total DNA was extracted after 5 days of culture in the absence or presence of Murabutide, and various concentrations (5.6, 28, 140, and 700 ng) were subjected to PCR amplification with primer pair GAG06-GAG04 to detect the HIV-1 gag gene. Cell equivalence was determined by amplification of the β-globin housekeeping gene. HIV-1 isolates from cultures of patients 3 and 4 presented X4 and R5 tropism, respectively.