Induction of IL-12Rβ2 mRNA is depressed in cells from HIV-infected individuals. PBMC from nine HIV+ patients and six healthy controls were cultured at 2 × 106/mL for 20 hours with PHA + TPA, anti-CD3 (OKT3), TPA + ionomycin (Io), or medium alone. RNA was extracted and competitive RT-PCR for IL-12Rβ2 was performed as described in Materials and Methods. (A) Representative samples of PHA + TPA-stimulated samples from each cohort are shown to demonstrate competitive IL-12Rβ2 RT-PCR. Near-equivalent competition is observed at 0.457 fg IL-12Rβ2 for the control sample and at 0.152 fg for the patient sample. (B) Densitometric values in arbitrary units were assigned by the ImageQuaNT program to the ethidium bromide-stained bands of wild-type and competitor fragments, which most closely approached equivalent competition. IL-12Rβ2 quantities were computed through the following formula: Unknown Sample Quantity = (ImageQuaNT Sample Value)(Quantity of Competitor)/(ImageQuaNT Competitor Value). Data represent the means of values derived from nine patients and six controls (except patients stimulated with TPA + ionomycin, wheren = 2). The P values comparing patient versus control group IL-12Rβ2 expression and unstimulated (no stim) versus stimulated groups are listed and were calculated via an unpaired student’s t-test. n.s., not significant.
