Figure 2: Treatment with AT-RvD1 prior to disease onset reduces lymphocytic infiltration and counter-regulates inflammatory responses in female NOD/ShiLtJ mice.

Female NOD/ShiLtJ mice were treated with AT-RvD1 and a vehicle control with and without DEX as described in the Material and Methods. Submandibular glands (SMG) were then harvested and sectioned as described in the Material and Methods. Tissue sections were stained with H&E and visualized on a Leica DMI6000B Inverted Microscope. Sections were scored using a histopathological grading system as described in the Material and Methods. Each point in the graph represents an average of the grades taken at three levels from a single mouse SMG (i.e., top, middle, and bottom) with a dashed threshold line of grade 3 indicating severe lymphocytic infiltration in SMG. (A) Representative images showing a grading scale of 0–4 utilized to assess the degree of lymphocytic infiltration (highlighted in yellow circle). Groups are as follows: (B) Females at 12 wk, and (C) females at 20 wk. Results from N > 7 mice were used per condition and data are expressed as mean ± SD, with *P < 0.05 indicating a significant difference from vehicle, and ^#P < 0.05 indicating a significant difference from RvD1. Additionally, RNA was isolated from the SMG for qPCR analysis of the following: (D) Pro-inflammatory cytokine profile at 12 wk, (E) pro-inflammatory cytokine profile at 20 wk, (F) anti-inflammatory molecule profile at 12 wk and (G) anti-inflammatory molecule profile at 20 wk. Results from N = 6 mice were used per condition and data are expressed as mean ± SD, with *P < 0.05 indicating a significant difference from controls. Orange boxes indicate pro-inflammatory cytokines and blue boxes indicate anti-inflammatory molecules.