Decidual Th1/Th2&NK1/NK2 Phenotyping with 8-color Flowcytometry in Recurrent Miscarriage

Background: Th1/Th2 imbalance has been considered as a mechanism for recurrent miscarriage. NK1/NK2 paradigm has also been hypothesized to play important role in pregnancy. There is little result to show the in-vivo changes of Th1/Th2 and NK1/NK2 subsets simultaneously in the decidual tissues.

Objective: The purpose of this study was to seek a method to observe the type1 and type2 subpopulations within decidua from women with recurrent miscarriages and from women having elective pregnancy termination.

Methods: Mononuclear cells were isolated from grinded decidual tissues in recurrent miscarriage cases (6–11 weeks of gestation, n=5) and early pregnant women (6–11 weeks of gestation, n= 5). The 8-color flowcytometry was performed, to demonstrate the expression of CD3, CD4, CD8, CD56, CD16, viability extracellular markers and INF-gamma, IL-4 intracellular markers.

Results: Th1/Th2 and NK1/NK2 subsets were visualized at the same time with 8-color flowcytometry successfully. IFN-γ(+)IL-4(-) or IL-4(+)IFN-γ(-) cell numbers can be calculated within one sample in the meantime.

Conclusion: The 8-color flowcytometry can be adapted in the research of decidual mononuclear cells. Investigations on the immunological mechanisms of normal or abnormal pregnancy can be facilitated in this way. More trials are needed to validate the actual changes of decidual Th1/Th2&NK1/NK2 paradigm in recurrent miscarriage.