title: Induction of Chemokine Secretion and Monocyte Migration by Human Choroidal Melanocytes in Response to Proinflammatory Cytokines.
abstract: To determine to which extent inflammatory cytokines affect chemokine secretion by primary human choroidal melanocytes (HCMs), their capacity to attract monocytes, and whether HCMs are able to influence the proliferation of activated T cells.
  
  
  
  Primary cultures of HCMs were established from eyes of 13 donors. Human choroidal melanocytes were stimulated with IFN-γ and TNF-α or with supernatant from activated T cells (T-cell-conditioned media [TCM]). Gene expression analysis was performed by using microarrays. Protein levels were quantified with ELISA or cytometric bead array. Supernatants of HCMs were assessed for the capability to attract monocytes in a transwell plate. Proliferation of activated T cells was assessed in a direct coculture with HCMs by a [3H]-thymidine incorporation assay.
  
  
  
  Stimulation of HCMs with TCM or IFN-γ and TNF-α resulted in increased expression and secretion of CXCL8, CXCL9, CXCL10, CXCL11, CCL2, CCL5 and intercellular adhesion molecule 1. Vascular endothelial growth factor and monocyte migration inhibitory factor were constitutively expressed without changes in response to proinflammatory cytokines. Supernatants derived from unstimulated cultures of 10 HCM donors induced a high initial level of monocyte migration, which decreased upon stimulation with either TCM or IFN-γ and TNF-α. The supernatants from three HCM donors initially showed a low level of monocyte attraction, which increased after exposure to proinflammatory cytokines. Direct coculture of HCMs with T cells resulted in inhibition of T-cell proliferation.
  
  
  
  These results showed that normal and activated HCMs are immunologically active by secreting chemokines, and that HCMs are able to attract monocytes in addition to inhibiting T-cell proliferation.
authors.affiliation: University of Copenhagen, Department of Immunology and Microbiology, Eye Research Unit, Copenhagen, Denmark.
authors.forename: Tina
authors.initials: T
authors.lastname: Jehs
authors.affiliation: University of Copenhagen, Department of Immunology and Microbiology, Eye Research Unit, Copenhagen, Denmark.
authors.forename: Carsten
authors.initials: C
authors.lastname: Faber
authors.affiliation: University of Copenhagen, Department of Immunology and Microbiology, Eye Research Unit, Copenhagen, Denmark.
authors.forename: Maja S
authors.initials: MS
authors.lastname: Udsen
authors.affiliation: Department of Ophthalmology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Martine J
authors.initials: MJ
authors.lastname: Jager
authors.affiliation: Centre for Ophthalmology and Vision Sciences, Institute of Human Development, AV Hill Building, University of Manchester, Manchester, United Kingdom.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Clark
authors.affiliation: University of Copenhagen, Department of Immunology and Microbiology, Eye Research Unit, Copenhagen, Denmark.
authors.forename: Mogens H
authors.initials: MH
authors.lastname: Nissen
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002465:Cell Movement; D049109:Cell Proliferation; D054370:Chemokine CXCL9 / Q000378:metabolism*; D002829:Choroid / Q000378:metabolism / Q000473:pathology*; D017077:Culture Media, Conditioned; D016207:Cytokines / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D008544:Melanocytes / Q000473:pathology*; D008545:Melanoma / Q000378:metabolism / Q000473:pathology*; D008875:Middle Aged; D009000:Monocytes / Q000473:pathology*; D010784:Photomicrography; D014407:Tumor Cells, Cultured; D014604:Uveal Neoplasms / Q000378:metabolism / Q000473:pathology*; D055815:Young Adult; D000098943:Uveal Melanoma
chemical_list: D054370:Chemokine CXCL9; D017077:Culture Media, Conditioned; D016207:Cytokines