FIG. 1.

ps20 mRNA correlates with HIV spread. One million cells (clones, primary CD4 CD45RO cells, and H9/HUT 78 cells) were infected overnight with a virus dose standardized as to HIV Gag p24-CA concentration (2 ng 2044 strain or 4 ng NL4-3 strain). Expanded CD4 cells and clones were stimulated with allogeneic allophycocyanin-PHA-IL-2 for 5 days prior to infection and cultured at 2 × 10⁵/ml in 30 IU/ml IL-2 postinfection. Mean peak p24-CA levels (day 8) in three biological replicate cultures are shown. qRT-PCR results for ps20 per population were determined in triplicate at the time of infection, and the mean number of ps20 molecules per cell is shown. (a) Comparison of NP and P counterpart clones from donors 8, 134, and 86 infected with the 2044 (X4) strain. (b) For ex vivo results, CD4 CD45RO⁺ T cells were infected with 2044 and then cultured in 30 IU/ml IL-2. For expanded results, purified CD4⁺ CD45RO⁺ T cells were expanded by two rounds of stimulation with allogeneic PBMC-PHA-IL-2 (see Materials and Methods) and then infected with 2044 and maintained in 30 IU/ml IL-2 postinfection. (c) Comparison of H9 versus HUT 78 immortalized cells infected with NL4-3 or 2044 strains. (d) A statistical correlation of peak p24-CA levels versus ps20 mRNA molecules per cell for each population was derived on GraphPad PRISM software. The two-tailed P value derived by use of the nonparametric Spearman's correlation is shown. The goodness-of-fit R² value derived by linear regression analysis is shown.