Micropreparative Cell Lysate Fractionation in Studying the Effect of Natural Killer Cells on Phenotype, Migration and Apoptosis of Trophoblast Cells in vitro
A. V. Korenevsky, Yu. P. Milyutina, S. K. Bochkovsky, A. A. Oshkolova, O. N. Bespalova, S. A. Selkov, D. I. Sokolov
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引用次数: 0
Abstract
Natural killer (NK) cells are a population of innate immune
cells that have cytotoxic functions and are involved in protecting
the body from viruses and transformed cells. Placental development
is largely determined by the interaction of decidual NK cells and
trophoblast cells. During pregnancy, NK cells accumulate around
trophoblast cells, while regulating trophoblast proliferation, migration
and invasion through the secretion of cytokines and growth factors.
The trophoblast, in turn, secretes chemokines and expresses ligands
for NK cell adhesion receptors. Thus, functional regulation of trophoblast
and NK cells is reciprocal. Despite intensive research, the role
of NK cells and methods for correcting their functional activity
in reproduction remain controversial. The aim of this study was
to assess the effect of NK cell lysate protein fractions on the
phenotype, migration and apoptosis of trophoblast cells in an in
vitro model experiment, using a new methodological approach. Chromatographic
separation yielded six fractions with different protein cargoes.
It was found that CD105 (endoglin) expression on the surface of JEG-3
trophoblast cells after their cultivation in the presence of high-
(< 250 kDa) or low-molecular-weight (< 45 kDa) fractions of
NK-92 whole-cell lysate was reduced compared to spontaneous expression,
with the relative count of trophoblast cells with a CD105+ phenotype
being also lowered. In addition, one of the low-molecular-weight
fractions decreased TRAIL-R2 receptor expression by trophoblast
cells. The high-molecular-weight fractions did not enable trophoblast
cells to migrate completely through a semi-permeable membrane, with
the area occupied by the migrated cells not exceeding the baseline
control area. Moreover, the high-molecular-weight fraction containing
the TGFβ dimer increased p-SMAD2/3 level in trophoblast cells 1
h after co-culture, followed by a decrease in the level of this
phosphorylated form after 2 or more hours, and also elevated procaspase-3
level one day after co-culture. The data obtained hypothetically
reflect the possible behavior of chorion cells under the influence
of collapsing NK cells in the event of their death under both normal
and pathological conditions caused by viral and bacterial infections,
as well as other stress factors leading to reproductive pathology.
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