Figure 6.

Changes in the frequency of regulatory lymphocyte cell populations inside the MLN of NOD mice following acute microparticle administration. Animals from the acute study cohort (See timeline in Figure 4A) were euthanized at 1 or 3 days following test article administration. Single cell suspensions from resected MLN were then assessed by flow cytometry for Treg and Breg frequency. A non-significant downward trend is seen in Treg frequency in MLN of mice treated with Ins-RT-NP vs. saline. Breg frequency exhibited an upward trend in mice treated with Ins-RT-NP, becoming statistically-significant significant compared to control test articles by the time of the third injection (n = 9, ANOVA p = 0.0262). Tukey's multiple comparisons determined p = 0.0215 between the saline and Ins-RT-NP treatment group. The Treg/Breg ratio was similarly significantly-different with the three-injection treatment (ANOVA p = 0.0035, post test p = 0.0025 for saline vs. Ins-RT-NP). Statistical significance is displayed as * for p < 0.05 and ** for p < 0.01 as assessed by ANOVA. Data are combined from the outcomes in two independent animal cohorts examined at different times in the study.