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Isolation of Uterine Innate Lymphoid Cells for Analysis by Flow Cytometry
09:02

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Published on: October 14, 2021

Uterine natural killer cells severely decrease in number at gestation day 6 in mice.

Akiko Takashima1, Fumio Ishikawa, Taku Kuwabara

  • 1Department of Obstetrics and Gynecology, Toho University Medical Center, Sakura Hospital, Sakura-shi, Chiba, Japan.

Biology of Reproduction
|September 13, 2013
PubMed
Summary

Uterine natural killer (uNK) cells, crucial for pregnancy, do not proliferate in the uterus after implantation. Instead, new natural killer (NK) cells migrate from the bloodstream, mature, and support pregnancy.

Keywords:
guinea pigsimplantationmicenatural killer cellpregnancyrodents (ratsuterusvoles)

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Area of Science:

  • Reproductive immunology
  • Maternal-fetal interface immunology
  • Innate immune cell dynamics during pregnancy

Background:

  • Uterine natural killer (uNK) cells increase significantly after implantation, but their origin remains debated.
  • The role of pre-existing uNK cells in the virgin uterus during early pregnancy is unclear.
  • Understanding uNK cell dynamics is vital for reproductive health and managing pregnancy complications.

Purpose of the Study:

  • To investigate the contribution of resident uNK cells versus circulating NK cell precursors to the uNK cell population expansion during early pregnancy.
  • To characterize the changes in uNK cell phenotype, maturation, and function throughout early gestation.
  • To elucidate the molecular mechanisms, including chemokine and cytokine signaling, involved in uNK cell regulation.

Main Methods:

  • Analysis of uterine leukocyte subsets in BALB/c mice at different gestation days (gd 0, 6, and 12) using flow cytometry.
  • Assessment of cell surface marker expression (CD11b, Gr-1, CD27, CD127, B220) to determine cell identity and activation status.
  • Evaluation of gene and protein expression for key signaling molecules (CXCL12, CXCR4, IL-15, Interferon-gamma, IL-15 receptor alpha).

Main Results:

  • A dramatic decrease in CD11b⁻/Gr-1⁻ cells, including uNK cells, was observed at gd 6, indicating selective depletion.
  • Resident uNK cells at gd 0 were CD11b⁻ and decreased significantly by gd 6, with recovery by gd 12.
  • uNK cells at gd 12 showed increased expression of maturation/activation markers (CD11b, CD27, CD127, B220) and produced higher levels of Interferon-gamma compared to gd 0.
  • Expression patterns of CXCL12/CXCR4 and IL-15 signaling molecules changed dynamically during early pregnancy.

Conclusions:

  • Resident uNK cells present before implantation do not significantly contribute to the post-implantation surge in uNK cell numbers.
  • The expansion of the uNK cell population is primarily driven by the migration of NK cell precursors from the bloodstream.
  • These newly arrived cells mature within the uterus and produce Interferon-gamma, essential for supporting a successful pregnancy.