FIG. 8.

Protozoal DNA stimulates enhanced steady-state levels of transcripts for inflammatory cytokines and iNOS in macrophages. RNA was collected from macrophages stimulated with 1 ng of lipopolysaccharide (LPS) per ml without or with 50 U of IFN-γ per ml or 25 μg of unmethylated or methylated E. coli, T. cruzi, T. brucei, or B. bovis DNA per ml, IFN-γ and 10 μg of polymyxin B sulfate per ml. Following RNA isolation and DNase treatment, RNA was analyzed by RT-PCR. Panels A to C and panels D to F represent two independent experiments. PCR products were semiquantified by nonsaturating densitometry and normalized to actin, and each sample was evaluated relative to the medium control. (B) IL-12 p40 analysis of samples in panel A; (C) iNOS analysis of samples in panel A; (E) IL-12 p40 analysis of samples in panel D; (F) iNOS analysis of samples in panel D. A third experiment gave a similar pattern of results.