Fig. 7.

Effect of TLR9 antagonist treatment on immune cells in Tlr9^+/+ NOD mice. (a) Regulatory T cells (Foxp3⁺ Tregs) in pancreatic islets. Immune cells were extracted from pancreatic islets isolated from the female progeny (5–6 weeks old) of Tlr9^+/+ NOD mice treated with TLR9 antagonist ODN or control ODN during pregnancy (n = 3 pregnant dams/group). Cells were stained with monoclonal antibodies to CD3, CD4 and Foxp3. The percentage of Foxp3⁺CD4 T cells is shown after gating for CD3⁺ cells (p = 0.235; n = 6 mice for both groups). (b) Reduced naive (CD44⁻CD62L⁺) CD4⁺ T cells in the spleen of the female progeny (5–6-weeks of age) from TLR9 antagonist-treated pregnant NOD mice. Splenocytes from these mice were isolated and stained with monoclonal antibodies to CD3, CD4, CD44 and CD62L. The percentage of naive CD4⁺ T cells is shown after gating for CD3⁺CD4⁺ T cells (p = 0.0011; n = 7 mice for both groups). (c) Increased memory (CD44⁺CD62L⁻) CD4⁺ T cells in the spleen of the female progeny (5–6 weeks old) from TLR9 antagonist-treated pregnant NOD mice. Splenocytes from these mice were isolated and stained with monoclonal antibodies to CD3, CD4, CD44 and CD62L. The percentage of memory CD4⁺ T cells is shown after gating for CD3⁺CD4⁺ T cells (p = 0.0106; n = 7 mice for both groups). Two independent experiments were carried out. Data are expressed as means (SD). Data were analysed by two-tailed unpaired Student’s t test. *p < 0.05, **p < 0.01