FIG.3.
Expression of ROS and gp91 phox immunoreactivity in porcine neutrophils. (A to D) NBT reduction due to ROS activity. The arrows indicate formazan-positive neutrophils removed from 1326/28Φr-inoculated pigs (B), F98-inoculated pigs (C), and 1326/28Φr-protected pigs after F98 challenge (D). Formazan was not observed in NBT reactions used to examine blood neutrophils taken from uninfected pigs (A) unless they were challenged with bacteria in vitro (not shown). (E to H) Confocal microscope images showing gp91 (green) immunofluorescence associated with the cell membrane (arrows) in neutrophils isolated from uninfected protected pigs (E). In neutrophils isolated from 1326/28Φr-inoculated pigs (F), F98-inoculated pigs (G), and 1326/28Φr-protected pigs following F98 challenge (H), gp91 immunofluorescence appears to be increased throughout the cytoplasm (arrows). (I to J) Overlaid images of gp91 (green) and propidium iodide nuclear stain (red) activities in neutrophils removed from 1326/28Φr-protected pigs following F98 challenge (I) or neutrophils isolated from unprotected F98-infected pigs (J). Scale bars = 10 μm.