Mikhailova V A, Khokhlova E V, Bazhenov D O, Agnaeva A O, Kozyreva A R, Bespalova O N, Selkov S A, Sokolov D I
Federal State Budgetary Scientific Institution Research Institute of Obstetrics, Gynecology, and Reproductology Named After D.O. Ott, Saint Petersburg, Russia.
Federal State Budgetary Scientific Institution Research Institute of Experimental Medicine, Saint Petersburg, Russia.
Cytotechnology. 2019 Jul 17;71(4):861-871. doi: 10.1007/s10616-019-00331-4.
The aim of this research was to assess the proliferative activity of Natural Killer Cells (NK cells) from Peripheral Blood Mononuclear Cells (PBMCs) in the presence of trophoblast cells in women with a history of recurrent miscarriages. We examined the peripheral blood of women with recurrent miscarriage in the proliferative (n = 12) or secretory (n = 13) phase of their menstrual cycle, and pregnant women with a history of recurrent miscarriage at 6-7 weeks of their current pregnancy (n = 14). Controls were fertile non-pregnant women in the proliferative (n = 11) or secretory (n = 13) phase of their menstrual cycle, and pregnant women at 6-7 weeks of a physiologically normal pregnancy (n = 20). We used IL-2 as a factor maintaining PBMCs viability during long-term culturing. We established that culturing in the presence of IL-2 contributed to an increase in the number of CD56+CD16- NK cells and to a decrease in the number of CD56+CD16+ NK cells from PBMCs compared with these numbers before culturing in both healthy women and in women with recurrent miscarriage. After culturing of PBMCs in the presence of trophoblast cells and IL-2 (compared with culturing without trophoblast cells), the intensity of Ki-67 expression by NK cells was reduced in the whole NK cell population (CD3-CD56+), and in the CD56+CD16- and CD56+CD16+ populations of NK cells in women with recurrent miscarriage and in healthy controls. The intensity of CD56 expression was reduced in the presence of trophoblast cells and IL-2 in non-pregnant women with recurrent miscarriage in the secretory versus the proliferative phase of the menstrual cycle.
本研究的目的是评估有复发性流产史女性的外周血单个核细胞（PBMC）中的自然杀伤细胞（NK细胞）在滋养层细胞存在时的增殖活性。我们检查了处于月经周期增殖期（n = 12）或分泌期（n = 13）的复发性流产女性的外周血，以及当前怀孕6 - 7周且有复发性流产史的孕妇（n = 14）。对照组为处于月经周期增殖期（n = 11）或分泌期（n = 13）的未孕有生育能力女性，以及生理正常怀孕6 - 7周的孕妇（n = 20）。我们使用白细胞介素-2（IL - 2）作为在长期培养过程中维持PBMC活力的因子。我们发现，与健康女性和复发性流产女性培养前相比，在IL - 2存在下培养会导致PBMC中CD56 + CD16 - NK细胞数量增加，CD56 + CD16 + NK细胞数量减少。在滋养层细胞和IL - 2存在下培养PBMC（与无滋养层细胞培养相比）后，复发性流产女性和健康对照组中整个NK细胞群体（CD3 - CD56 +）以及NK细胞的CD56 + CD16 - 和CD56 + CD16 + 群体中，NK细胞的Ki - 67表达强度降低。在月经周期分泌期与增殖期的有复发性流产的未孕女性中，滋养层细胞和IL - 2存在时CD56表达强度降低。
