Induction of IL-12 mRNA by a Multistimulus Panel Is Impaired in Cells From HIV-Infected Subjects
| Conditions . | p40 . | p35 . | ||
|---|---|---|---|---|
| HIV− . | HIV+ . | HIV− . | HIV+ . | |
| −/− | 1 | 1 | 1 | 1 |
| −/S aureus | 112.3 | 49.1 | 5.7 | 3.2 |
| −/LPS | 72.2 | 7.4 | 2.6 | 1.7 |
| IFN-γ/LPS | 96.9 | 98.5 | 6.3 | 4.5 |
| −/OK432 | 117.9 | 75.6 | 5.1 | 2.6 |
| −/LTA | 90.7 | 27.1 | 1.9 | 1.2 |
| IFN-γ/LTA | 183.5 | 48.0 | 9.9 | 6 |
| IL-4/S aureus | 125.6 | 44.1 | 7.9 | 3.7 |
| Conditions . | p40 . | p35 . | ||
|---|---|---|---|---|
| HIV− . | HIV+ . | HIV− . | HIV+ . | |
| −/− | 1 | 1 | 1 | 1 |
| −/S aureus | 112.3 | 49.1 | 5.7 | 3.2 |
| −/LPS | 72.2 | 7.4 | 2.6 | 1.7 |
| IFN-γ/LPS | 96.9 | 98.5 | 6.3 | 4.5 |
| −/OK432 | 117.9 | 75.6 | 5.1 | 2.6 |
| −/LTA | 90.7 | 27.1 | 1.9 | 1.2 |
| IFN-γ/LTA | 183.5 | 48.0 | 9.9 | 6 |
| IL-4/S aureus | 125.6 | 44.1 | 7.9 | 3.7 |
PBMCs were isolated from 13 HIV+ patients and 10 control subjects. PBMCs (2.5 × 106) from each donor were stimulated in1 mL culture medium for 16 h ± IFN-γ or IL-4 followed by 4 hours of stimulation with S aureus Cowan (SAC), LPS, OK432, LTA, or medium alone. The same 2 mL Ultraspec was used for all subjects within a condition, thus each PCR band represents pooled RNA from 2.5 × 106 cells from each of the 13 or 10 individuals of the group. RT-PCR was performed for the detection of p40, p35, TNF-α, and HPRT message. Densitometric values in arbitrary units were assigned to the ethidium bromide-stained bands by the ImageQuaNT program and these were normalized to HPRT values. Fold-increase values quantitate the increase of p40 and p35 mRNA expression after stimulation compared with unstimulated cells (−/−).