title: Association of CD14 and TLR4 with LPS-stimulated human normal skin fibroblasts in immunophenotype changes and secretion of TGF-β1 and IFN-γ.
abstract: OBJECTIVE
  We attempted to explore the association of CD14 and TLR4 with LPS-stimulated human normal skin fibroblasts in immunophenotype changes and secretion of TGF-β1 and IFN-γ, and to expand the current knowledge of the mechanisms that underlie LPS-induced scar formation.
  
  
  METHODS
  We randomized the human normal skin fibroblasts cultured in vitro into four groups. The expression profile of immune phenotypes was determined by immunohistochemical staining. Ultrastructure of cells was observed by use of transmission electron microscopy. Secretion status of TGF-β1 and IFN-γ was inspected using ELISA assay.
  
  
  RESULTS
  Compared with group A, the expressions of α-SMA and α1 (I) procollagen in groups B, C, D were lower, and it in group D were the lowest in all groups. The cells in group A were diversification under the electron microscope, and the ratio of the nuclear to plasma of the fibroblasts was large, with unregular nuclear membrane, more Golgi apparatus, rough endoplasmic reticulum, and microfilament and canaliculus appeared. The ultrastructure of the fibroblasts in group B, C, D was spindle and the nuclear was large, with regular nuclear membrane, more Golgi apparatus, rough endoplasmic reticulum. ELISA assay indicated that the secretion of TGF-β1 markedly lowered in groups B, C, D in comparison to group A, with the most marked decline observed in group D. Interestingly, we found significantly increased IFN-γ secretion in groups B, C, D (P < 0.05), with the latter group showing the most notable increase (P < 0.01).
  
  
  CONCLUSIONS
  These data suggest that both combined and isolated use of CD14 and TLR4 significantly reduce α-SMA expression levels, the number of α1 (I) pro-collagen positive cells, and TGF-β secretion, while substantially increased IFN-γ secretion. The reduction and increase are especially notable when pretreating with CD14 and TLR4 combined. Here we thus draw a conclusion that both CD14 and TLR4 are associated with the immunophenotype changes and secretion of TGF-β1 and IFN-γ in LPS-stimulated human normal skin fibroblasts.
authors.affiliation: Department of Burn and Plastic Surgery, First Hospital Affiliated to General Hospital of PLA Beijing 100048, China.
authors.forename: Hongming
authors.initials: H
authors.lastname: Yang
authors.affiliation: Department of Burn and Plastic Surgery, First Hospital Affiliated to General Hospital of PLA Beijing 100048, China.
authors.forename: Juncong
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Burn and Plastic Surgery, First Hospital Affiliated to General Hospital of PLA Beijing 100048, China.
authors.forename: Yihe
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Burn and Plastic Surgery, First Hospital Affiliated to General Hospital of PLA Beijing 100048, China.
authors.forename: Quan
authors.initials: Q
authors.lastname: Hu
mesh_terms: D002460:Cell Line; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D020796:Receptor, Platelet-Derived Growth Factor alpha / Q000494:pharmacology*; D012867:Skin / Q000187:drug effects / Q000378:metabolism*; D051197:Toll-Like Receptor 4; D053773:Transforming Growth Factor beta1 / Q000378:metabolism*
chemical_list: D008070:Lipopolysaccharides; D051197:Toll-Like Receptor 4; D053773:Transforming Growth Factor beta1; D007371:Interferon-gamma; D020796:Receptor, Platelet-Derived Growth Factor alpha
keywords: CD14; Lipopolysaccharide; collagen; fibroblast; toll like receptor 4