Fig. 6

Flow analysis of immune cells; and ELISA of IL-2 and IFN-γ production. a Representative flow plots of leukocyte distributions of (from left to right) (1) non surgery, non-IS treatment control (NN AMC); (2) IS treated RO transplant recipient (NN TP IS); and 3) foxn1−/− mutant RO transplant recipient (Nude TP) RD rats. Uppermost row isolates lymphocyte from non-lymphocyte proportions of leukocyte population of each, based on characteristic forward and side scatter distribution. Where leukocytes are further distinguished into lymphocytes (“lymphos”) and non-lymphocyte leukocytes (“non-lymphos”). Lower row quantifies CD11bc+ myeloid populations from ‘non-lympho’ proportion of leukocytes from each respective immune status and treatment category. b Mean population profile of myeloid proportion of leukocytes (CD11 bc +) across immune status and treatment categories. Note immunocompetent sham surgery category is included. n = 3 each category. Both immune deficient (nude TP) and NN TP IS group demonstrating significantly higher relative populations of myeloid CD11 b/c+ cells than immune competent control or sham groups. TP NO IS group not represented. c Serum IL-2 levels of single surgery cohort approximately 1 week (8 days) and 7 weeks (43 days) post-surgery, Surgery groups: NN TP IS n = 1, NN TP NO IS n = 2, and NN Sham, n = 2. Mean values are given where n > 1. d and e Comparison of serum IFN-γ levels across immunocompetent surgery modalities 1–2 days and 3–4 weeks post-surgery and in non-surgery, non-treatment age matched RD control group. 1–2 days post-surgery: N = 4 for each group. 3–4 weeks post-surgery, for each group NN AMC n = 4; NN TP IS n = 7; NN TP NO IS n = 6; NN Sham n = 6. Statistical analysis performed using ordinary one-way ANOVA with Dunnet multiple comparisons test. TP NO IS expressing greatest average IFN-γ. (though not statistically significant compared to NN TP IS). The sham group expressed the lowest overall IFN-γ levels; likely a result of homeostatic, immune modulatory rebound from the initial inflammation inciting event of invasive surgery, well within range of the competent immune response. f Serum IFN-γ levels from 1–2 days to 3–4 weeks post-surgery (n = 4 individuals from each surgery modality spanning 2–3 surgery cohorts each group measured at indicated time points). g Proportion of IFN-γ producing cells across surgery categories at 3–4 weeks post-surgery and h Representative proportions of IFN-γ producing immune cells across treatment categories of immune competent RD rats. e Table combines relative populations of IFN-γ producing cells normalized to non-surgery control. Across treatment categories, interrogates relationship between T-cell and NK populations upstream and serum IFN-γ levels downstream. X-axis values represent ratios of indicated categories compared to a normalized to immune competent, non -surgery control (NN AMC) representative of immune homeostatic levels