Figure 1.
Phenotypic composition of T cell subsets in the ileum and mesenteric lymph nodes (MLN) of normal rats during day of life (DOL) 0 to DOL 4.
The frequencies of T cell subsets including CD3+, CD4+, CD8+, CD4+CD8+, CD4+Foxp3+, and double positive CD4+CD8+Foxp3+ cells, and the comparisons between DOL 0 and DOL 4 were shown by representative flow cytometry plots in ileum (1A–a) and mesenteric lymph node (1B–a) of newborn rats. The bars reflect the percentage of cells, shown as means ± SE, N = 9 pups at each DOL. Comparisons were made between on DOLs 1, 2, 3, or 4 compared with DOL0 in the ileum (1A–b) or MLN (1B–b) of newborn rats, respectively. *p<0.05, **p<0.01, and ***p<0.001.
Figure 2.
Effect of breast milk or formula feeding with or without Lactobacillus reuteri DSM 17938 on the frequency of Foxp3+ T cells in the ileum and mesenteric lymph node (MLN) of rats during DOL 0 and DOL 4.
Newborn rats were fed immediately after birth for 4 days. (a) and (c): Changes of single CD4+Foxp3+ Tregs (%) in the ileum (a) and MLN (c); (b) and (d): changes of double CD4+CD8+Foxp3+ T cells in the ileum (b) and MLN (d). The percentage (%) of cells is represented as the mean ± SE of N = 9 observations (pups) in each DOL of each feeding group (total N = 45 for each feeding group).* Dam-fed +17938 vs. Dam-fed, **p<0.01, ***p<0.001. # Formula vs. Dam-fed, # p<0.05, ## p<0.01. &: Formula-fed +17938 vs. Formula-fed, & p<0.05, && p<0.01.
Figure 3.
Effect of Lactobacillus reuteri DSM 17938 on survival, CD3+ T cells and Foxp3+ T cells in the ileum of rats with NEC.
(a) Survival (%) of different groups of rat pups (dam-fed, NEC, and NEC+17938). (b) Changes of % CD3+ T cells of lymphocytes in different groups. (c) Immunohistochemical staining of CD3+ cells in rat pup ileum, 200 × magnification. (d) Number of CD3+ T cells per 5 villi in IHC. (e) Percentage of CD4+Foxp3+Tregs in each group on day 4 of NEC. (f) Percentage of double positive CD4+CD8+Foxp3+ T cells. The numbers represent percent of T cells (means ± SE). Each dot represents one animal in each group in the figures b, e, and f. Dam-fed: N = 15, NEC: N = 13, NEC+17938: N = 20.
Figure 4.
Effect of Lactobacillus reuteriDSM 17938 on CD3+, CD4+ and Foxp3+ T cells in the mesenteric lymph nodes of rats with NEC.
(a): changes of CD3+ T cells of lymphocytes, (b) % of CD4+ T cells/CD3+ T cells, (c) % of CD4+Foxp3+, and (d) double positive CD4+CD8+Foxp3+ cells in the mesenteric lymph nodes in different groups. The numbers represent percent of T cells (mean ± SE). Each dot represents one animal in each group in the figures. Dam-fed: N = 15, NEC: N = 13, NEC+17938: N = 20.