Figure 1.
In situ detection of U. parvum in placental and fetal tissues.
Representative 2D projection images (600 x magnifications) of placental choriodecidual junction (A), vitelline membrane (B), fetal intestine (C), and fetal lung (D). Scale bars are equivalent to 100 µm. U. parvum (red) was initially labeled with rabbit polyclonal antibody as previously described [69], [71]. Cell nuclei (blue) were labeled with DAPI. Long arrows (A and B) are highlighting U. parvum engulfed by polymorphonuclear cells. Block arrows (D and E) are highlighting epicellular U. parvum.
Table 1.
Lesion scoring criteria for maternal inflammatory response [32].
Figure 2.
Representative images of metritis.
Sham inoculated (Control) and representative lesions in U. parvum infected C57BL/6 mice (B, mild metritis) and in U. parvum infected BALB/c mice (C and D, more extensive metritis). Scale bars are equivalent to 100 µm. Arrows are highlighting neutrophilic infiltrates.
Figure 3.
Maternal inflammatory response defined by the degree of chorioamnionitis in U. parvum infected BALB/c and C57BL/6 dams.
Distribution of raw maternal inflammatory response (MIR) placental scores using the scoring criteria of Redline et al. [32] summarized in Table 1. Median scores are demarcated with a horizontal line in each graph. Data are a compilation of 3 independent experiments. Accompanying H & E stained photomicrographs of MIRS stages 1–3. Scale bars are equivalent to 100 µm. Arrows are highlighting neutrophilic infiltrates.
Table 2.
Fetal inflammatory response criteria modified from Redline 2006 [32].
Figure 4.
Fetal inflammatory response (FIR) in U. parvum infected BALB/c and C57BL/6 mice.
Distribution of raw FIR scores as described in Table 2. Median scores are demarcated with a horizontal line in each graph. Data are a compilation of 3 independent experiments. Accompanying H & E stained sections are representative images of fetal pathology in umbilical vessels, brain, lung, liver, and intestine. Scale bar is equivalent to 100 µm. Arrows are highlighting neutrophilic infiltrates.
Figure 5.
Profiling of placental inflammatory mediators in sham inoculated BALB/c and C57BL/6 mice. (
A) Normalized gene expression values are expressed as 2−ΔCt [72]. (B) Production of IL-10 expressed as mean (pg/gm weight of tissue) ± SD. Values represent 5 biological replicates from 3 independent experiments.
Figure 6.
Placental inflammatory profiles in U. parvum infected BALB/c and C57BL/6 mice. (
A) Fold change (2−ΔΔCt ) is relative to normalized Ct values from sham inoculated controls, which was determined by the method of Schmittgen and Livak [72]. Values represent the mean 2−ΔΔCt ± SD of 5 biological replicates from 3 independent experiments.* Indicates a significant difference (P<0.01), which was determined by Student’s t test using 2−ΔCt data. (B) Mean IL-10± SD expressed as pg/gm weight of tissue of 5 biological replicates from 3 independent experiments. ** Indicates a significant difference (P<0.03).
Table 3.
Predominant BALB/c and C57BL/6 response profile to U. parvum intrauterine infection.