FIGURE 7.

Adoptive transfer (Tx) of FoxP3+ T regulatory cells (T_reg) arrests EAU progression. (A) Adoptive transfer (Tx) of unfractionated T cells (1 × 10⁶/mouse). Lymph node cells (submandibular and superficial cervical LN) were collected from adult P28-P42 3A9 TCR or dTg HEL/TCR mice and adoptively transferred (i.v.) to P21 mice (n = 6–7/treatment group). Control mice received no cells (“No”; n = 4). Eyes were taken for histology at P30. ^∗∗∗p < 0.001 on a 95% level of confidence. (B,C) Tx (i.v.) of enriched FoxP3+CD25+ T_reg cells from spleen and submandibular, superficial cervical, axillary, and inguinal LN, passed through a CD4+CD25+ regulatory T cell isolation kit (see section “T_reg Cell Isolation”) [B: CD127+; C: CD127-; 1 × 10⁶/mouse; bead-based CD127 negative selection (18)] arrests EAU progression. T_reg were isolated from 3A9 TCR mice aged P50-70 (naïve T_reg), and dTg mice of the same age (antigen-experienced T_reg), and adoptively transferred (i.v.) to P21 dTg mice. **p < 0.01, *p < 0.05 on a 95% level of confidence. In control experiments (D) FoxP3-CD25-CD4+ effector cells from P50-70 3A9 controls (n = 2) and dTg mice of the same age (n = 8) were administered to P21 mice (3 × 10⁶/mouse), and their effect on EAU progression evaluated. Fundus images of P20 mice were taken on the day before Tx (pre-Tx) and repeated 9 days later (post-Tx), on P30. Eyes were assessed for inflammation using fundoscopy (B–D), and histology (A) (see “Materials and Methods”) ^∗∗p < 0.01 on a 95% level of confidence.