Lung tissue is the primary tissue colonized by Y. pestis during pneumonic infections. Because the lungs reside in the thoracic cavity covered by other organs and bone, we again used B6(Cg)-Tyrc-2J/J mice to increase the probability
of detecting signal from lung tissue. In some isolated cases, radiance was detected from the abdomen and from feces at 6 hpi (data not shown). This signal was not detected at any latter time points and presence of abdominal or fecal signal did not appear to alter the course of infection in the animals where it was detected. Very little light was detected in the mice at 24 hpi, at which time some mice showed signal #Talazoparib solubility dmso randurls[1|1|,|CHEM1|]# from different regions in the neck or on the head (Figure 6A). At 48 hpi, light was detected in all animals, mainly from the mid
and upper thorax (Figure 6B). Radiance spread and intensity increased considerably at 72 hpi, Lonafarnib solubility dmso a time at which all mice showed pronounced signs of disease. Immediately after imaging at 72 hpi, one of the four mice in the group was sacrificed and dissected to determine the source of light. The lungs were determined to be the source of luminosity from the thorax, and light from this organ was confirmed to be unique to IN infections as animals infected using other routes (e.g. ID, Figure 6C) did not show signal from the lungs. Additionally, we observed that IN-inoculated animals showed signal from the tip of the nose (visible in Figure 6C) indicating that bacteria were present at the site of inoculation at 72 hpi. Upon dissection of the lungs, we noticed that part of the organ was necrotic in appearance; imaging of isolated lungs showed that the necrotized tissue produced higher levels of signal (Figure 6D) in comparison to other areas of the lung. While Figure 6C and 6D show data from only one mouse, we performed this experiment
multiple times and in all cases we made the same observations mentioned above (data not shown). Figure 6 BLI of C57BL/6J mice infected subcutaneously with Δ caf1 Δ psaA Y. pestis carrying the pGEN- luxCDABE vector. (A) Mice were inoculated with ~200 CFU of the double mutant. Images correspond to infected animals at different time points post inoculation (shown in hpi). A color bar serves as a reference for the radiance scale (p/sec/cm2/sr) VAV2 used to standardize all images. (B) Images of superficial cervical lymph nodes, spleen and liver (from one of the mice shown in A) imaged individually after dissection. Luminescence was detected only from lymph nodes, imaged in an individual scale of radiance with a Min = 2.28e6 and Max = 4.27e7. (C) Transformed values (ln) of the mean radiance per group from the neck (left) and abdomen (right) from animals infected with Yplux + (gray circles) and YpΔcaf1ΔpsaA lux + (white circles), as determined by measurements from regions of interest (ROI) of images from two independent experiments. A dotted line depicts background radiance levels.