Down-regulation of HLA-G boosted natural killer cell-mediated cytolysis in JEG-3 cells cultured in vitro

Fertil Steril. 2008 Dec;90(6):2398-405. doi: 10.1016/j.fertnstert.2007.10.076. Epub 2008 Feb 20.

Abstract

Objective: To determine how decidual natural killer (NK) cells interact with fetal trophoblasts in vitro.

Design: Prospective study.

Setting: University hospitals and IVF units.

Patient(s): Not applicable.

Intervention(s): Not applicable.

Main outcome measure(s): An adenovirus vector containing small interfering RNA (siRNA) specifically targeting the human lymphocyte antigen-G (HLA-G) gene was constructed and applied to diminish HLA-G mRNA expression. The steady-state levels of HLA-G messenger RNA (mRNA) were then checked by reverse transcriptase-polymerase chain reaction (RT-PCR) and protein levels by Western blot analysis. The NK-mediated cell cytotoxicity in the siRNA treated cells was studied by application of a nonradioactive cytotoxicity assay.

Result(s): Steady-state levels of HLA-G mRNA and protein were significantly diminished by the targeting siRNA. In cells where HLA-G expression was thus reduced, a significant increase in NK cell-mediated lysis occurred.

Conclusion(s): These results indicate that the recombinant adenoviral vectors used were efficient tools for studying HLA-G function. More important, this study reveals an important immunoprotective function for HLA-G in controlling NK cell-mediated lysis of trophoblasts, cells whose role in mediating normal pregnancy is important.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics
  • Cell Line, Tumor
  • Coculture Techniques
  • Cytotoxicity Tests, Immunologic
  • Cytotoxicity, Immunologic*
  • Down-Regulation
  • Genetic Vectors
  • HLA Antigens / genetics
  • HLA Antigens / metabolism*
  • HLA-G Antigens
  • Histocompatibility Antigens Class I / genetics
  • Histocompatibility Antigens Class I / metabolism*
  • Humans
  • Killer Cells, Natural / immunology*
  • Prospective Studies
  • RNA Interference
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / metabolism
  • Transfection
  • Trophoblasts / immunology*
  • Trophoblasts / pathology

Substances

  • HLA Antigens
  • HLA-G Antigens
  • Histocompatibility Antigens Class I
  • RNA, Messenger
  • RNA, Small Interfering