engleski

The expression of cytolytic mediators in decidual mononuclear cells in first trimester of pregnancy

The main focus of our current study has been to investigate the levels of gene and protein expression of various cytolytic mediators, such as perforin, granulysin, TRAIL and Fas-L, which are present in leukocyte cell subpopulations of first trimester pregnancy decidua. Decidual lymphocytes were separated from freshly isolated decidual mononuclear cells (DMC) after an 18-hour culture at 37oC. Cell subpopulations of interest (such as CD56+ NK cells, NK T-cells (NKT), CD3+ T-lymphocytes, CD14+ macrophages and CD1a+ immature dendritic cells) were then separated by cell sorting or magnetically. Total RNA was isolated using the Trizol method (Invitrogen). Subsequently, mRNA levels for granulysin, perforin, FasL and TRAIL were detected by quantitative RT-PCR using Taqman probes (AB 7300 Real-Time PCR system). Our preliminary quantitative RT-PCR results suggest that, of the four cytolytic mediators examined, granulysin is the most predominantly expressed gene in decidual NK and NKT cells. A similar pattern is observed in other decidual cell subpopulations, such as immature dendritic cells and macrophages. Furthermore, after the 18 hour culture of decidual lymphocytes with K562-HLA-G stably transfected cell line, the protein expression of granulysin and other cytolytic mediators (perforin, FasL, TRAIL) is markedly reduced and their intracellular distribution is re-arranged, whereas the mRNA levels appear to increase. In conclusion, although decidual lymphocytes at the maternal-foetal interface are primed to kill, they are at the same time under the efficient control of the trophoblast itself. Further experiments investigating these effects should bring us closer to understanding the finely tuned immune events occurring during human pregnancy. Acknowledgement: The experiments were financed by Croatian Ministry of Science, Education and Sport, Grants No. 0620402-0376 and No. 062-620402-0379.

Decidual mononuclear cells; pregnancy; granulysin

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