engleski

Granulysin and perforin expression in human early pregnancy decidua

Problem: Granulysin is a cytolytic/apoptotic molecule especially high expressed on human decidual NK cells and activated, but not resting cytolytic T cells. Perforin is calcium-dependent pore forming protein. The aim of the study was to analyze the gene expression of granulysin and perforin in various population of decidual mononuclear cells, intracellular localization of granulysin and perforin in decidual lymphocytes, peripheral blood lymphocytes and decidual CD56+ cells. Methods of study: Decidual mononuclear cells (DMC) were obtained by enzymatic digestion of decidual tissue and density gradient centrifugation. Peripheral blood lymphocytes were obtained by density gradient centrifugation. CD56+ cells were purified from the suspension of a non-adherent DMC fraction using positive magnetic separation after 18-hour culture at 37°C. Granulysin and perforin protein expression were analyzed using confocal microscopy. Granulysin and perforin mRNA expression were determined by quantitative RT -PCR using Taqman probes (AB 7300 Real-Time PCR). Results: In comparison to perforin, granulysin mRNA is present at significantly higher levels in NK, NKT, immature dendritic cells and macrophages, but not in T lymphocytes. In decidual lymphocytes granulysin is mostly stored in the granules near the cell membrane, whereas perforin is stored in more centrally positioned granules. Granulysin is quantitatively more colocalized with perforin in peripheral blood lymphocytes compared to decidual lymphocytes. Conclusion: Abundant expression of cytolytic molecules granulysin and perforin in the decidual lymphocytes at the fetal-maternal interface indicates their important role in the early pregnancy outcome. Lower colocalization with perforin in decidual lymphocytes may indicate the fine balance in lymphocyte activation/suppression, needed for the survival of embryo. Acknowledgement: The investigation was financed by Croatian Ministry of Science, grant No. 0620402-0376 to Dr. D. Rukavina.

Granulysin; NK cells; pregnancy

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