Table 3. Composition of organ populations is altered in control vs. TEMPOL-fed EAE animals.
Cells were isolated 10 days after EAE induction in animals on control or TEMPOL feed for 14 days prior to immunization. Analyses were performed on flow cytometry data by selecting the live cells based on forward and side scatter, then selecting the CD45+ leukocyte population, then subsequently gating the described populations as a percentage of CD45+ cells. Values represent average results of 2 pooled experiments (n = 13 mice total per feed group) for lymphoid organs ±SD, and 8 animals per group, brain and spinal cord pooled 2 to 1 (n = 4 per group).
| Control Average % |
TEMPOL Average % |
P value | |
|---|---|---|---|
| Lymph node | |||
| B cells (CD19+) | 55.6 ±3.4 | 54.6 ±4.1 | 0.441 |
| Myeloid cells (CD11b+CD11c−) | 1.1 ±0.6 | 1.8 ±0.8 | 0.051 |
| Dendritic cells (CD11c+CD11b−) | 1.3 ±0.2 | 1.3 ±0.2 | 0.681 |
| Myeloid DCs (CD11b+CD11c+) | 2.0 ±0.5 | 2.3 ±0.3 | 0.045* |
| NK cells (NK1.1+) | 4.0 ±0.7 | 4.5 ±0.5 | 0.043* |
| T cells (CD3+) | 37.9 ±4.4 | 38.2 ±2.9 | 0.538 |
| % CD4+ T cells | 53.3 ±3.3 | 46.7 ±2.3 | <0.001* |
| % CD8+ T cells | 40.1 ±2.1 | 46.4 ±2.2 | <0.001* |
| % CD4+CD25+Foxp3+ T cells | 9.7 ±2.6 | 12.3 ±2.5 | 0.027* |
| Spleen | |||
| B cells (CD19+) | 36.3 ±7.3 | 34.2 ±6.3 | 0.719 |
| Myeloid cells (CD11b+CD11c−) | 26.1 ±8.0 | 29.4 ±5.0 | 0.442 |
| Dendritic cells (CD11c+CD11b−) | 1.5 ±0.3 | 1.4 ±0.3 | 0.259 |
| Myeloid DCs (CD11b+CD11c+) | 3.1 ±1.4 | 2.9 ±0.4 | 0.858 |
| NK cells (NK1.1+) | 4.9 ±1.5 | 4.7 ±2.4 | 0.305 |
| T cells (CD3+) | 19.7 ±8.0 | 17.6 ±4.3 | 0.918 |
| % CD4+ T cells | 56.6 ±3.6 | 50.7 ±4.2 | 0.001* |
| % CD8+ T cells | 32.1 ±3.2 | 37.7 ±4.0 | 0.001* |
| % CD4+CD25+Foxp3+ T cells | 10.2 ±2.2 | 8.8 ±1.1 | 0.045* |
| CNS – brain & spinal cord | |||
| # cells recovered (x106) | 1.8 ±0.2 | 1.0 ±0.1 | |
| CD45 high leukocytes | 62.0 ±8.8 | 28.1 ±6.5 | 0.006* |
| CD11b | 32.7 ±8.6 | 53.7 ±5.5 | 0.027* |
| CD3 | 57.7 ±8.0 | 39.3 ±5.5 | 0.031* |
Mann-Whitney Rank Sum test with uncorrected p values are shown for each subset analyzed comparing control to TEMPOL fed animals, *p < 0.050. Bolded values are those withstanding Bonferroni correction for six populations analysed in the spleen and lymph node (p < 0.0083) and correction for four populations analyzed in the CNS (p < 0.0125).