Over 1.5 million individuals suffer from cornea vascularization due to genetic and/or environmental factors， compromising visual acuity and often resulting in blindness. Current treatments of corneal vascularization are limited in efficacy and elicit undesirable effects including， ironically， vision loss. To develop a safe and effective therapy for corneal vascularization， adeno-associated virus (AAV) gene therapy， exploiting a natural immune tolerance mechanism induced by human leukocyte antigen G (HLA-G)， was investigated. Self-complementary AAV cassettes containing codon optimized HLA-G1 (transmembrane) or HLA-G5 (soluble) isoforms were validated in vitro. Then， following a corneal intrastromal injection， AAV vector transduction kinetics， using a chimeric AAV capsid， were determined in rabbits. One week following corneal trauma， a single intrastromal injection of scAAV8G9-optHLA-G1?+?G5 prevented corneal vascularization， inhibited trauma-induced T-lymphocyte infiltration (some of which were CD8)， and dramatically reduced myofibroblast formation compared to control treated eyes. Biodistribution analyses suggested AAV vectors persisted only in the trauma-induced corneas; however， a neutralizing antibody response to the vector capsid was observed inconsistently. The collective data demonstrate the clinical potential of scAAV8G9-optHLA-G to safely and effectively treat corneal vascularization and inhibit fibrosis while alluding to broader roles in ocular surface immunity and allogenic organ transplantation.