Down-regulation of HLA-G boosted natural killer cell-mediated cytolysis in JEG-3 cells cultured in vitro.
OBJECTIVE
To determine how decidual natural killer (NK) cells interact with fetal trophoblasts in vitro.


METHODS
Prospective study.


METHODS
University hospitals and IVF units.


METHODS
Not applicable.


METHODS
Not applicable.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Sun
Li Li
LL
Department of Gynecology and Obstetrics, Tangdu Hospital, The Fourth Military Medical University, Xi'an, PR China.
Han
Yibing
Y
Chen
Jian Hui
JH
Yao
Yuan Qing
YQ
D000256:Adenoviridae / Q000235:genetics; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D003601:Cytotoxicity Tests, Immunologic; D003602:Cytotoxicity, Immunologic; D015536:Down-Regulation; D005822:Genetic Vectors; D006680:HLA Antigens / Q000235:genetics / Q000378:metabolism*; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology*; D011446:Prospective Studies; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D014162:Transfection; D014327:Trophoblasts / Q000276:immunology* / Q000473:pathology
D006680:HLA Antigens; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I; D012333:RNA, Messenger; D034741:RNA, Small Interfering