Phenotypic characterization of NKT-like cells and evaluation of specifically related cytokines for the prediction of unexplained recurrent miscarriage

Wafaa S Khalaf,Mohammad R.A Mahmoud,Walid F Elkhatib,Hany R Hashem,Wafaa E Soliman

doi:10.1016/j.heliyon.2021.e08409

Abstract

ProblemImmune system dysregulation is a major cause of unexplained recurrent miscarriage (URM). Women with URM need screening for their pregnancy microenvironment and immune regulators, to prevent spontaneous abortion. Method of studyIn this study we evaluated NKT-like cell subsets in peripheral venous blood of women with URM using flow cytometry. The expression levels of specifically related Th1 cytokines (IFN-γ and IL-2), Th2 cytokine (IL-4), and Th17 cytokines (IL-17), were measured using enzyme-linked immunosorbent assay. ResultsThe percentage of CD16+CD56+NKT-like (Double Positive NKT-like; DPNKT-like) cell subset, and the levels of IL-2 and IFN-γ were significantly elevated in blood of non-pregnant and pregnant patients with URM compared with the healthy control groups, and these parameters were significantly increased after pregnancy in the same patients with URM. Based on the prevalence of the candidate immunological factors in patients with URM, the prognostic significance of the NKT-like cell subsets, IFN-γ and IL-2 profiles were evaluated as potential predictors of URM. A cut-off point of 2.55% for DPNKT-like cell subset in the blood and cut-off values of 39.5 and 20.5 pg/ml for the levels of IFN-γ and IL-2, respectively could be used for the prediction of the risk of spontaneous abortion. To the best of our knowledge, this is the first study that described the prognostic significance of the aforementioned immunological parameters before and after pregnancy, and highlighted the correlation of NKT-like cells and the candidate Th1 cytokines with pregnancy loss in women with URM. ConclusionsDPNKT-like cells, IFN-γ and IL-2 patient profiles could be used as markers to predict the risk of miscarriage in patients with URM.

Highlights

Recurrent miscarriage is defined as two or more successive pregnancy losses within the first trimester [1]
To assess the percentage of CD3þT cells and different subsets of NKTlike cells (CD3þCD56þCD16þ (DPNKT-like cells), CD3þCD56þCD16À, and CD3þCD56ÀCD16þ NKT-like cells) in the Peripheral venous blood (PVB) of unexplained recurrent miscarriage (URM) patients and the healthy control group, flow cytometric analysis of these cell subsets was performed, using PVB samples isolated from 90 non-pregnant patients with URM and 90 healthy individuals (Figure 1A and B)
The evaluation of different NKT-like cell subsets showed that the percentage of DPNKT-like cells subsets in non-pregnant women with a history of URM was significantly greater (P

Summary

Introduction

Recurrent miscarriage is defined as two or more successive pregnancy losses within the first trimester [1]. Recurrent miscarriage affects about 15% of all pregnancies, and up to 50% of women who experienced recurrent abortions with no defined etiology [1]. The underlying etiology of recurrent miscarriage is highly debated, there is considerable evidence suggesting a role for various immune system regulators in recurrent miscarriage [2]. NKT-like cells have a major influence, controlling both the innate and the adaptive pregnancy immune responses [3, 4, 5, 6]. NKT-like cells were defined using the co-expression of CD3 and CD56 cell markers [7]. Vitelli-Avelar and colleagues further classified NKT-like cells with the additional inclusion of CD16 as CD3þCD56þCD16þ (DPNKT-like cells), CD3þCD56þCD16À, and CD3þCD56ÀCD16þ [11]

Methods

Results

Discussion

Conclusion