Figure 3.

pTregs show a higher susceptibility to convert into exFoxp3 cells after transfer into dry eye hosts. pTregs (CD4⁺CD25⁺Nrp-1⁻) and tTregs (CD4⁺CD25⁺Nrp-1⁺) from transgenic dry eye disease (DED) mice (Foxp3-GFP×R26-RFP) were adoptively transferred into control and DED transplant recipients. (A and B) Draining lymph nodes were harvested at day 14 post-transplantation. Single cell suspensions were analyzed by flow cytometry. Frequencies of exFoxp3 cells (GFP⁻RFP⁺) from control and DED hosts that received (A) pTregs (59.02 ± 0.92% vs. 78.26 ± 0.69%, n = 4, ***p < 0.0001 unpaired two-tailed Student’s t test) or (B) tTregs was assessed. (C) Representative flow cytometry plots showing frequencies of IL-17 and IFNγ-expressing exFoxp3 cells in draining lymph nodes from control and dry eye hosts. (D) Nrp1⁺ tTregs or Nrp1⁻ pTregs from hosts with accepted allografts were adoptively transferred to dry eye hosts, and graft survival was assessed for 8 weeks (n = 10, χ² = 0.6087, p = 0.4353).