Abstract
Purpose :
To assess the immunological effects of Rho-Kinase Inhibitor (Ripasudil) on murine corneal transplantation.
Methods :
Donor corneas (C57BL6) were transplanted onto BALB/c recipient mice. Placebo instillation (P group), 0.04% Ripasudil (R1 group), 0.4% Ripasudil (R2 group), 2.0% Ripasudil (R3 group) were instilled three times a day post transplantation immediately. Draining lymph nodes (dLNs) were harvested at day 14 post-transplantation (n=5 mice/group). We analyzed the frequencies of CD4+CD25+Foxp3+regulatory T cells and MHCIIhiCD11c+ mature dendritic cells (DCs) in the host dLNs using flow cytometry. The expression of Foxp3, CTLA-4 on Treg and MHCII of DCs were analyzed by mean fluorescence intensity (MFI).
Results :
Treg was significantly decreased in the R3 group compared to the P group (14.95% in P group, 14.77% in R1 group, 15.22% in R2 group, 13.60% in R3 group, p = 0.0096). MHCIIhiCD11c+ mature DCs were significantly decreased in the R2 and R3 groups compared to the P group (63.22% in P group, 59.52% in R1 group, 56.78% in R2 group, 55.12% in R3 group, p = 0.003). The MFI expression of Foxp3 in Tregs showed a significant decrease in the R3 group (P: 2398, R1: 2332, R2: 2347, R3: 2165, p = 0.0243) and the expression of MHC2 in DCs showed a significant decrease in the R3 group (P: 10889, R1: 10142, R2: 10034, R3: 7666, p = 0.0023). There was no significant difference in the expression of CTLA-4 in each group.
Conclusions :
Ripasudil instillation could inhibit maturation of dendritic cells in corneal transplantation.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.