title: Cytokine expression in human dermal fibroblasts stimulated with eosinophil cationic protein measured by protein array.
abstract: BACKGROUND
  Eosinophil cationic protein (ECP) was reported previously to be involved in allergic inflammation with cytotoxic activity. On the other hand, recent studies showed that ECP did not induce cell death but inhibited the growth of cancer-derived cells. Our previous study indicated that human ECP enhanced differentiation of rat neonatal cardiomyocytes and stress fiber formation in Balb/c 3T3 mouse fibroblasts, while the effects of human ECP on human fibroblasts are unknown.
  
  
  OBJECTIVE
  The present study was performed to determine the effects of human ECP on cytokine expression in human fibroblasts by protein array.
  
  
  METHODS
  The effects of recombinant human ECP (rhECP) on normal human dermal fibroblasts (NHDF) were examined by assaying cell growth. Furthermore, cytokine expression of NHDF stimulated by ECP, which could influence cell growth, was evaluated by protein array.
  
  
  RESULTS
  ECP was not cytotoxic but enhanced the growth of NHDF. The peak rhECP concentration that enhanced the cell counts by 1.56-fold was 100 ng/mL, which was significantly different from cultures without ECP stimulation (ANOVA/ Scheffe's test, P < 0.05). Array analyses indicated that ciliary neurotrophic factor (CNTF), neutrophil-activating peptide (NAP)-2, and neurotrophin (NT)-3 were significantly upregulated in NHDF stimulated with 100 ng/mL ECP compared to those without stimulation.
  
  
  CONCLUSIONS
  ECP is not cytotoxic but enhances the growth of NHDF. CNTF, NAP-2, and NT-3 were suggested to be involved in enhancing the growth of NHDF. These findings will contribute to determination of the role of ECP in allergic inflammation.
authors.affiliation: Department of Pathophysiology-Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
authors.forename: Takamaro
authors.initials: T
authors.lastname: Sato
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Soga
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yamaguchi
authors.forename: Michio
authors.initials: M
authors.lastname: Meguro
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Maeda
authors.forename: Joji
authors.initials: J
authors.lastname: Tada
authors.forename: Takayuki
authors.initials: T
authors.lastname: Otani
authors.forename: Masaharu
authors.initials: M
authors.lastname: Seno
authors.forename: Shogo
authors.initials: S
authors.lastname: Takashiba
mesh_terms: D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis*; D047150:Eosinophil Cationic Protein / Q000276:immunology / Q000378:metabolism* / Q000494:pharmacology; D005347:Fibroblasts / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D040081:Protein Array Analysis; D011994:Recombinant Proteins; D012867:Skin / Q000166:cytology / Q000276:immunology
chemical_list: D016207:Cytokines; D011994:Recombinant Proteins; D047150:Eosinophil Cationic Protein; C479220:RNASE3 protein, human