Differential regulation and function of the Fas/Fas ligand system in human trophoblast cells

Biol Reprod. 2002 Jun;66(6):1853-61. doi: 10.1095/biolreprod66.6.1853.

Abstract

Trophoblast rejection, which is characterized by increased apoptosis, is mediated by T helper (Th)-1, or proinflammatory, cytokines, whereas Th-2, or anti-inflammatory, cytokines confer immune protection and facilitate implantation. We investigated the role of both types of cytokines on the expression and function of the Fas/Fas ligand (FasL) apoptotic pathway in trophoblast cells. First-trimester human trophoblast primary-culture cells as well as A3 and HTR/8 trophoblast cell lines were treated with proinflammatory cytokines such as interferon-gamma (IFN-gamma) and tumor necrosis factor alpha (TNFalpha) and with the anti-inflammatory cytokines interleukin (IL)-6 and IL-10. Sensitivity to Fas-mediated apoptosis was measured using an activating anti-Fas monoclonal antibody. Cell viability was evaluated using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) and CellTiter 96 assay. Fas/FasL mRNA and protein expression levels were determined using reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. Trophoblast cells normally express FasL, but low levels of Fas, and they are resistant to Fas-mediated apoptosis. IFN-gamma and TNFalpha promote Fas expression and sensitivity, whereas IL-6 and IL-10 increase the resistance of trophoblast cells to Fas-mediated apoptosis. Furthermore, IL-10 treatment activates FLICE-like inhibitory protein (FLIP), a downstream inhibitor of Fas apoptotic signaling. Although trophoblast cells express Fas, susceptibility to Fas does not necessarily correlate with its expression. In this study, we demonstrate that Th-2 cytokines increase the resistance of trophoblast cells to Fas-mediated apoptosis either by inhibiting Fas expression or by inducing FLIP activation. This "trophoblast-cytokine-Fas/FasL triad" determines the ability of the Fas/FasL system to regulate trophoblast viability and, consequently, the success or failure of pregnancy.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Apoptosis
  • Blotting, Western
  • CASP8 and FADD-Like Apoptosis Regulating Protein
  • Carrier Proteins / metabolism
  • Cells, Cultured
  • Cytokines / pharmacology*
  • Fas Ligand Protein
  • Female
  • Gene Expression Regulation*
  • Gestational Age
  • Humans
  • Interferon-gamma / pharmacology
  • Interleukin-10 / pharmacology
  • Interleukin-6 / pharmacology
  • Intracellular Signaling Peptides and Proteins*
  • Membrane Glycoproteins / genetics*
  • Membrane Glycoproteins / physiology*
  • Pregnancy
  • Reverse Transcriptase Polymerase Chain Reaction
  • Th1 Cells / metabolism
  • Th2 Cells / metabolism
  • Trophoblasts / physiology*
  • Tumor Necrosis Factor-alpha / pharmacology
  • fas Receptor / genetics*
  • fas Receptor / physiology*

Substances

  • CASP8 and FADD-Like Apoptosis Regulating Protein
  • CFLAR protein, human
  • Carrier Proteins
  • Cytokines
  • FASLG protein, human
  • Fas Ligand Protein
  • Interleukin-6
  • Intracellular Signaling Peptides and Proteins
  • Membrane Glycoproteins
  • Tumor Necrosis Factor-alpha
  • fas Receptor
  • Interleukin-10
  • Interferon-gamma