Novel immunotherapeutic effects of topically administered ripasudil (K-115) on corneal allograft survival

• Published in: Scientific reports 2020-11, Vol.10 (1), p.19817-19817, Article 19817
• Main Authors: Inomata, Takenori, Fujimoto, Keiichi, Okumura, Yuichi, Zhu, Jun, Fujio, Kenta, Shokirova, Hurramhon, Miura, Maria, Okano, Mikiko, Funaki, Toshinari, Sung, Jaemyoung, Negishi, Naoko, Murakami, Akira
• Format: Article
• Language: English
• Subjects: 631/250/1854; 692/699/3161; 692/699/3161/3163; Administration, Topical; Allografts; Angiogenesis; Animals; CD11b antigen; CD11c antigen; CD4 antigen; CD45 antigen; Cornea; Cornea - drug effects; Cornea - metabolism; Corneal Transplantation; Dendritic cells; Enzyme inhibitors; Flow cytometry; Graft Rejection; Humanities and Social Sciences; Immunohistochemistry; Immunology; Inflammation; Interleukin 17; Isoquinolines - therapeutic use; Kinases; Lymph nodes; Lymphocytes T; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; mRNA; multidisciplinary; Opacity; Polymerase chain reaction; Protein kinase inhibitors; Science; Science (multidisciplinary); Sulfonamides - therapeutic use; Survival; Transplantation, Homologous - methods; Vascularization; Wound healing; γ-Interferon
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-76882-w

Corneal allograft survival is mediated by the variety of immunological reactions and wound healing process. Our aim was to explore the effects of topical administration of ripasudil, a selective Rho-associated coiled-coil protein kinase inhibitor, on corneal allograft survival. Ripasudil was administered to mice thrice a day after allogeneic corneal transplantation. Corneal graft survival, opacity, neovascularization, re-epithelization, immune cell infiltration, and mRNA levels of angiogenic and pro-inflammatory factors in the grafted cornea and draining lymph nodes (dLNs) were evaluated with slit-lamp microscopy, immunohistochemistry, flow cytometry, and polymerase chain reaction. Graft survival was significantly prolonged with lower graft opacity and neovascularization scores in 0.4% and 2.0% ripasudil-treated groups, and mRNA levels of angiogenic and pro-inflammatory factors in ripasudil-treated grafted corneas were reduced. Moreover, 0.4% and 2.0% ripasudil reduced CD45 + -infiltrated leukocyte frequency, Cd11b and Cd11c mRNA levels, and the frequencies of mature dendritic cells, IFNγ-, and IL-17- producing CD4 + T cells in the dLNs of recipients. Re-epithelization rate of the grafted cornea was significantly higher in the 0.4% and 2.0% ripasudil groups than in the control. Topically applied ripasudil prolonged graft survival by downregulating neovascularization and inflammation factors, while promoting corneal re-epithelization, suggesting that ripasudil may be useful for suppressing immunological rejection in corneal transplantation.