title: Topical application of FTY720 and cyclosporin A prolong corneal graft survival in mice.
abstract: OBJECTIVE
  To investigate the effects of topical FTY720 and cyclosporin A (CsA) on allogeneic corneal transplantation in mice.
  
  
  METHODS
  A total of 75 BALB/c mice received corneal grafts from C57BL/6 donors. Recipients were treated with 0.1%, 0.3%, or 0.5% FTY720 ophthalmic gel or 1% CsA eye-drops after the graft (controls received no treatment). The number of cluster of differentiation (CD)4+ T cells and CD4+CD25+forkhead box P3 (Foxp3)+ regulatory (Treg) cell phenotypes were measured by flow cytometry. Cytokine mRNA expression in corneal grafts was analyzed by real-time quantitative PCR. CD4 + T cells and cytokines in corneal samples were identified by immunohistochemical staining.
  
  
  RESULTS
  Corneal graft survival was prolonged by treatment with topical 0.5% FTY720 (mean survival time [MST], 24.1±1.6 days) or 1% CsA eye-drops (MST 25.0±1.9 days) compared with controls (MST, 13.4±0.5 days; n=9, both p<0.01). Topical 0.5% FTY720 treatment significantly increased the percentages of CD4 + T (p<0.05) and Treg cells (p<0.01; n=5) in the cervical lymph nodes compared with controls. Transforming growth factor-β1 (TGF-β1) mRNA transcription in corneal grafts after topical 0.5% FTY720 increased (p<0.05, n=3), while interleukin-2 (IL-2) and interferon-γ (IFN-γ) mRNA expression in corneal grafts treated with 1% CsA decreased (p<0.01, p<0.05, respectively). These cytokine results were paralleled by similar immunohistochemical staining. Topical 0.5% FTY720 and 1% CsA treatment reduced the infiltration of CD4+ Tcells in the grafts.
  
  
  CONCLUSIONS
  Topical 0.5% FTY720 and 1% CsA can effectively prolong allogeneic corneal graft survival in mice. Treatment with topical 0.5% FTY720 increases the percentage of CD4+ T cells and the percentage of Treg cells in cervical lymph nodes. The 0.5% FTY720 increased TGF-β1 mRNA expression and decreases infiltration of CD4+ T cells in corneal grafts, while topical 1% CsA down-regulated the expression of IL-2 and IFN-γ.
authors.affiliation: Departments of Ophthalmology, Chinese PLA General Hospital, Beijing, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Liu
authors.forename: Jingjing
authors.initials: J
authors.lastname: Jiang
authors.forename: He
authors.initials: H
authors.lastname: Xiao
authors.forename: Xiaokui
authors.initials: X
authors.lastname: Wang
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Yubo
authors.initials: Y
authors.lastname: Gong
authors.forename: Dajiang
authors.initials: D
authors.lastname: Wang
authors.forename: Yifei
authors.initials: Y
authors.lastname: Huang
mesh_terms: D060433:Administration, Ophthalmic; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D003315:Cornea / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D016039:Corneal Transplantation; D016572:Cyclosporine / Q000008:administration & dosage* / Q000627:therapeutic use; D016207:Cytokines / Q000096:biosynthesis / Q000276:immunology; D000068876:Fingolimod Hydrochloride; D051858:Forkhead Transcription Factors / Q000096:biosynthesis; D005782:Gels; D006085:Graft Survival / Q000187:drug effects* / Q000276:immunology; D007166:Immunosuppressive Agents / Q000008:administration & dosage* / Q000627:therapeutic use; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009883:Ophthalmic Solutions; D011409:Propylene Glycols / Q000008:administration & dosage* / Q000627:therapeutic use; D013110:Sphingosine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000627:therapeutic use; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000187:drug effects; D014184:Transplantation, Homologous
chemical_list: D016207:Cytokines; D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse; D005782:Gels; D007166:Immunosuppressive Agents; D009883:Ophthalmic Solutions; D011409:Propylene Glycols; D016572:Cyclosporine; D000068876:Fingolimod Hydrochloride; D013110:Sphingosine