METHOD FOR NK-CELLS CYTOTOXIC ACTIVITY DETERMINATION Russian patent published in 2017 - IPC G01N33/53 

FIELD: medicine.

SUBSTANCE: method comprises isolation of a mononuclear cells fraction containing NK-cells from the peripheral blood. Incubation is carried out in the culture medium of mononuclears in the presence of target cells, followed by estimation of the number of dead cells by flow cytometry. At that, peripheral blood mononuclear cells containing NK-cells are incubated in the DMEM culture medium for 4 hours in the presence of trophoblast cells of the Jeg-3 line. Some of the trophoblast cells are incubated in DMEM culture medium without mononuclears for 4 hours. The number of nonviable and viable trophoblast cells incubated with peripheral blood mononuclear cells and the base number of nonviable and viable trophoblast cells incubated in the culture medium without peripheral blood mononuclear cells are then determined. The cytotoxic effect of NK-cells on trophoblast cells is determined by the formula: CE=(X1/X2)×100-(X1_base/X2_base)×100, where CE is the cytotoxic activity of NK-cells, X1 is the number of non-viable trophoblast cells incubated with peripheral blood mononuclear cells, X2 is the number of viable trophoblast cells incubated with peripheral blood mononuclear cells, X1_bases is the base number of non-viable trophoblast cells incubated in the culture medium without addition of mononuclear cells; X2_base is the base number of viable trophoblast cells incubated in the culture medium without addition of mononuclear cells.

EFFECT: use of this method allows to obtain quantitative evaluation of the NK-cells cytotoxic activity, which makes it possible to monitor the course of pregnancy.

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