title: Aspergillus fumigatus promotes T helper type 2 responses through thymic stromal lymphopoietin production by human corneal epithelial cells.
abstract: BACKGROUND
  Fungal keratitis is a major cause of blindness. To understand the mechanism of both innate and adaptive immunity in corneal infection is of great significance in the treatment and prevention of fungal keratitis. Our previous study concerned innate immunity. Here, we explored the potential role of thymic stromal lymphopoietin (TSLP) in adaptive immunity of fungal keratitis.
  
  
  METHODS
  Human corneal epithelial cells (HCECs) were stimulated with Aspergillus fumigatus hyphae (10(6) pieces per millilitre) with or without TSLP siRNA, and peripheral blood mononuclear cells (PBMCs) were cultured with or without TSLP. HCECs and PBMCs were co-cultured in a transwell system for various periods. Then we collected PBMCs and detected the proliferation and activation as well as T helper type 2 (Th2) differentiation by flow cytometry and quantitative real-time reverse transcription polymerase chain reaction. IgG and IgA levels in supernatants of PBMCs were measured by means of ELISA.
  
  
  RESULTS
  Thymic stromal lymphopoietin could induce a Th2 response in vitro, and the expression of TSLP was highly increased in HCECs stimulated with A. fumigatus hyphae. A. fumigatus-infected HCECs were capable of promoting human lymphocyte proliferation and activating human CD4(+) T cells, CD8(+) T cells and B cells by up-regulating the expression of activation marker CD69. Importantly, Th2 differentiation of CD4(+) T cells was induced during co-culture with A. fumigatus-infected HCECs in a transwell system. Interestingly, blockade of TSLP using siRNA prevented the proliferation and activation of lymphocytes as well as Th2 differentiation. We also detected an increased IgG level that was associated with TSLP.
  
  
  CONCLUSIONS
  These findings suggested that HCEC-derived TSLP has a key role in adaptive immune responses of fungal keratitis via skewing Th2 differentiation and promoting humoral immunity.
authors.affiliation: Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, China.
authors.forename: Luping
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, China.
authors.forename: Leyi
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, China.
authors.forename: Xinyi
authors.initials: X
authors.lastname: Wu
mesh_terms: D056704:Adaptive Immunity / Q000502:physiology*; D000328:Adult; D015703:Antigens, CD / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism; D001232:Aspergillus fumigatus / Q000502:physiology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002460:Cell Line; D018920:Coculture Techniques; D016207:Cytokines / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D019573:Epithelium, Corneal / Q000276:immunology / Q000378:metabolism / Q000382:microbiology*; D005260:Female; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D056724:Immunity, Humoral / Q000502:physiology; D007074:Immunoglobulin G / Q000097:blood; D037181:Lectins, C-Type / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008213:Lymphocyte Activation / Q000502:physiology; D008297:Male; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D018418:Th2 Cells / Q000276:immunology*; D014162:Transfection; D000094632:Thymic Stromal Lymphopoietin; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D016207:Cytokines; D007074:Immunoglobulin G; D037181:Lectins, C-Type; D034741:RNA, Small Interfering; D000094632:Thymic Stromal Lymphopoietin; D000099192:CD69 Antigens
keywords: Aspergillus fumigatus; Th2 response; adaptive immunity; human corneal epithelial cell; thymic stromal lymphopoietin