A Model of Corneal Graft Rejection in Semi-Inbred NIH Miniature Swine: Significant T-Cell Infiltration of Clinically Accepted Allografts

Abstract

The purpose of our study is to develop a pre-clinical model of corneal graft rejection in the semi-inbred NIH minipig as a model of human rejection. NIH minipigs received corneal allografts with MHC and minor mismatches, or minor mismatches alone. Clinical rejection was monitored, and major subsets of leukocytes and ingress of vessels were quantified post-mortem by automated digital methods. Spectratypes of recipient T-cell receptor β-subunit variable region (TRβV) were analyzed. The capacity of pig corneal endothelial cells to proliferate in vivo was assessed. Autografts (n = 5) and SLA(cc) to SLA(cc) allografts (minor mismatches, n = 5) were not rejected. Median graft survival of SLA(dd) and SLA(bb) allografts in SLA(cc) strain recipients (major and minor mismatches) was 57 (n = 10) and 67 (n = 6) days, respectively. Rejected grafts did not recover clarity in vivo, and corneal endothelial cells did not proliferate in organ culture after cryo-injury. There were significantly more leukocytes in clinically rejected versus accepted grafts (P < 0.0001) and in transplanted versus contralateral eyes (P < 0.0001). Numbers of T-cells were significantly greater in clinically accepted grafts versus autografts and in rejected grafts versus accepted (P < 0.005 for most subsets). There were significant differences in TRβV spectratype between graft groups in cornea, but not in draining lymph node or blood (P < 0.05). The NIH minipig offers a robust model of human rejection suitable for immunological or therapeutic studies. In particular, there is limited capacity for corneal endothelial repair in vivo, and histological evidence suggests that allosensitization of the recipient may develop in the absence of clinical rejection.