title: Partial characterization of a putative new growth factor present in pathological human vitreous.
abstract: BACKGROUND
  Several growth factors have been implicated in the development of proliferative eye diseases, and some of those are present in human vitreous (HV). The effects of HV on cellular responses which modulate proliferative cell processes were studied. This study describes the partial characterization of a vitreous factor activity which does not correspond to any of the previously reported growth factors in pathological HV.
  
  
  METHODS
  Vitreous humour was obtained from medical vitrectomies, from patients with PDR and PVR. The biological activity of the vitreous factor was determined by its ability to increase cytosolic calcium concentration ([Ca2+]i), increase production of inositol phosphates, and induce cell proliferation in the cell line EGFR T17. In some experiments other cell lines, such as NIH 3T3, 3T3-L1, FRTL5, A431, PC12, Y79, and GH3, were also employed. Measurement of [Ca2+]i in cell suspensions was performed using the fluorescent Ca2+ indicator fura-2. The activity of the factor present in HV was compared with other growth factors by means of: (a) [Ca2+]i mobilization pattern, (b) sequential homologous and heterologous desensitization of receptors, (c) effects of phorbol esters on their action, and (d) inactivation after treatment with different proteolytic enzymes.
  
  
  RESULTS
  The HV-induced cell proliferation and increases in [Ca2+]i concentration were characterized by a peculiar time pattern. The different approaches used ruled out its identity with PDGF, bFGF, EGF, TGF-beta, IGFs, TNF-alpha, NGF, and other compounds such as ATP, angiotensin I, and bradykinin. Vitreous factor actions are mediated by specific receptors apparently regulated by PKC. This factor is able to induce [Ca2+]i mobilization in most of the cell lines studied, indicating that its effects are not tissue specific.
  
  
  CONCLUSIONS
  These results suggest the presence of a growth factor activity in pathological HV which may be due to the presence of an undescribed growth factor in the eye.
authors.affiliation: Department of Medicine, Faculty of Medicine, University of Santiago de Compostela, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Pombo
authors.forename: L
authors.initials: L
authors.lastname: Bokser
authors.forename: X
authors.initials: X
authors.lastname: Casabiell
authors.forename: J
authors.initials: J
authors.lastname: Zugaza
authors.forename: M
authors.initials: M
authors.lastname: Capeans
authors.forename: M
authors.initials: M
authors.lastname: Salorio
authors.forename: F
authors.initials: F
authors.lastname: Casanueva
mesh_terms: D002118:Calcium / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003930:Diabetic Retinopathy / Q000209:etiology / Q000503:physiopathology / Q000601:surgery; D000431:Ethanol / Q000494:pharmacology; D005136:Eye Proteins / Q000502:physiology*; D005347:Fibroblasts / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D016257:Fura-2 / Q000031:analogs & derivatives / Q000378:metabolism; D006133:Growth Substances / Q000502:physiology*; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007295:Inositol Phosphates / Q000096:biosynthesis; D014407:Tumor Cells, Cultured / Q000378:metabolism; D014821:Vitrectomy; D018630:Vitreoretinopathy, Proliferative / Q000209:etiology / Q000503:physiopathology / Q000601:surgery; D014822:Vitreous Body / Q000502:physiology*
chemical_list: D005136:Eye Proteins; D005456:Fluorescent Dyes; D006133:Growth Substances; D007295:Inositol Phosphates; C049925:fura-2-am; D000431:Ethanol; D002118:Calcium; D016257:Fura-2