Desiccating Stress-induced Dry Eye Disease is a High-Risk Factor for Corneal Allograft Rejection

Abstract

Purpose: We established a murine transplantation model in dry eye recipients to investigate the effect of desiccating stress-induced ocular surface inflammation as a high-risk condition in corneal transplantation.

Methods: Dry eye disease (DED) was induced by exposing BALB/c mice to a controlled environment chamber (desiccating stress) for 14 days, and then corneal allograft transplantation was performed using wild-type C57BL/6 or transgenic CD45.1⁺ donor corneas. Graft opacity, neovascularization, and survival were assessed for 8 weeks after transplantation. 24 hours after transplantation we analyzed the frequencies and maturation status of donor- and recipient-derived antigen-presenting cells (APCs) in the draining lymph nodes using flow cytometry. T cell proliferation was assessed in a mixed lymphocyte reaction assay using BrdU incorporation. We determined the in vivo T cell response using a delayed-type hypersensitivity (DTH) assay. T cell allosensitization was measured using ELISPOT assay.

Results: Desiccating stress significantly increased graft rejection, graft opacity and neovascularization. We observed significantly higher frequencies of both donor APCs (0.61 ± 0.03% vs. 0.22 ± 0.03%, n=8/group, p=0.0009) and recipient APCs in the DLNs of DED recipients compared to controls (room air) (1.01 ± 0.13% vs. 0.62 ± 0.06%, n=8/group, p=0.02). Donor-derived APCs in DED recipients expressed increased MHC II (maturation marker) and co-stimulatory molecules (CD80 and CD86). T cell proliferation and T cell allosensitization were increased in DED recipients.

Conclusions: These results demonstrate that pre-existing DED in corneal graft recipients enhances allosensitization, and thus is a high-risk factor for allograft rejection.