Figure 1
Herbimycin A inhibits BCG- and SP-A-BCG-induced production of nitric oxide. BCG were collected by centrifugation, and then suspended in PBS. SP-A (20 μg/5 × 105 BCG) or buffer was added, and the mixtures incubated for 30 min at 37°C. The BCG (B) or SP-A-BCG (B/S) complexes were pelleted, resuspended in medium, and added to RBMM (5 × 105) in 24 well plates at an MOI of 1. One-half of the cells from each treatment (BCG or SP-A-BCG) were exposed to herbimycin A (HA) at a concentration of 100 nM. Cells plus mycobacteria were incubated for 24 hr in serum-free DMEM. The spent culture medium was removed at 24 hr, and nitrate/nitrite levels were measured using the Griess reagent. Results are the average ± S.D. for triplicate determinations, and are representative of four separate experiments. *p < .001 for B/S compared to BCG; **p < .001 for B+HA compared to BCG; ***p < .001 for B/S + HA compared to B/S.