engleski

Granulysin mediates long term cytolytic activity of the first trimester pregnancy decidual NK cells

Problem:We investigated the granulysin mediated cytotoxicity of early pregnancy decidual CD56 positive (dCD56+) cells against NK-sensitive K562 cell line. Methods of study: CD56+ cells were positively selected in a magnetic field from a non-adherent decidual mononuclear cell fraction after 18-hrs culture at 37°C. Granulysin expression in unstimulated or K562 pulsed dCD56+ cells was analyzed using flow cytometry, immunohistochemistry or confocal microscopy. The cytotoxicity of CD56+ cells against K562 cells was investigated using PKH-26 red 2-hrs and 18-hrs cytotoxicity assay. Results: Approximately 90% of unstimulated decidual NK cells express granulysin in their cytoplasmatic granules located near the cell membrane. After the contact with K562 cells, the distribution of granulysin+ granules aquired more diffuse distribution. Although dCD56+ cells lyse K562 cells in 2-hrs and 18-hrs cytotoxicity test, the addition of anti-granulysin antibody decreased only 18-hrs cytotoxicity. Conclusion: Granulysin seems to be a long term cytolytic mediator employed by dCD56+ cells. Acknowledgement: The experiments are financed by Croatian Ministry of Science, grant No. 0620402-0376 and EMBIC project, European FP6 project No. 512040, NoE, LSHM-CT-2004-512040

granulysin; cytotoxicity; early pregnancy; K562

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