Table_2_Recurrent Implantation Failure May Be Identified by a Combination of Diagnostic Biomarkers: An Analysis of Peripheral Blood Lymphocyte Subsets.docx

BackgroundRecurrent implantation failure (RIF) is a challenge during assisted reproductive technology (ART). In the present study, potential diagnostic biomarkers for the immune status of peripheral blood lymphocyte subsets in patients with RIF were analyzed, with the aim of identifying novel biomarkers that may predict RIF.
MethodsA total of 41 participants, including 21 women with RIF and 20 fertile controls, were included in the present study. Functional analysis was performed and the cytokine status of natural killer (NK), T, CD8+ T, T helper (Th), and γδ T cells which are lymphocyte subsets in peripheral blood was measured using flow cytometry. Binary logistic regression analysis adjusted for T follicular helper 1 (Tfh1), Tfh2, Tfh17, and early NK cells was performed to determine the relationship between the peripheral blood lymphocyte subsets and RIF. Potential diagnostic biomarkers were assessed by logistic regression analysis and receiver operating characteristic curves.
ResultsThere were significantly more Tfh1, Tfh17, and NK cells in the RIF group compared with the control group (all P < 0.05). However, the percentage of T, regulatory T (Tregs), and Tfh2 cells, as well as early inhibitory NK cells, was significantly lower in the RIF group compared with the control group (all P < 0.05). Following logistics regression analysis, Treg, Tfh17, and early inhibitory NK cells exhibited significant differences between the two groups. Combination diagnosis using these 3 biomarkers had a higher area under the curve of 0.900 (95% confidence interval: 0.808–0.992, P < 0.001) in the RIF group compared with that in the control group.
ConclusionT, Tregs, Tfh1, Tfh2, Tfh17, NK cells, and early inhibitory NK cells may play important regulatory roles in embryo implantation. The combination of 3 molecular markers (Treg, Tfh17, and early inhibitory NK cells) could provide a high diagnostic value for women with RIF, thus providing novel potential biomarkers for RIF in ART. The present findings could provide a reference either for the clinical treatment of patients with RIF or for future large, well-designed studies.

背景：复发性植入失败（RIF）是辅助生殖技术（ART）实施过程中的一大临床挑战。本研究针对复发性植入失败患者外周血淋巴细胞亚群的免疫状态潜在诊断标志物展开分析，以期筛选出可用于预测RIF的新型生物标志物。
方法：本研究共纳入41名受试者，其中21名复发性植入失败患者与20名生育功能正常的对照个体。开展功能分析，并采用流式细胞术（flow cytometry）检测外周血淋巴细胞亚群——自然杀伤（NK）细胞、T细胞、CD8+T细胞、辅助性T（Th）细胞及γδT细胞——的细胞因子状态。以滤泡辅助性T1（Tfh1）、Tfh2、Tfh17及早期NK细胞为校正变量，进行二元logistic回归分析，以明确外周血淋巴细胞亚群与RIF之间的关联。通过logistic回归分析与受试者工作特征曲线（receiver operating characteristic curves, ROC）评估潜在诊断标志物的诊断价值。
结果：与对照组相比，复发性植入失败组的Tfh1、Tfh17及NK细胞占比显著升高（所有P<0.05）。但T细胞、调节性T（Treg）细胞、Tfh2细胞及早期抑制性NK细胞的占比，在复发性植入失败组中显著低于对照组（所有P<0.05）。经logistic回归分析后，Treg细胞、Tfh17细胞与早期抑制性NK细胞在两组间存在显著差异。联合应用这3种生物标志物进行诊断时，其曲线下面积（area under the curve, AUC）可达0.900（95%置信区间：0.808~0.992，P<0.001），诊断价值显著优于对照组。
结论：T细胞、Treg细胞、Tfh1、Tfh2、Tfh17、NK细胞及早期抑制性NK细胞可能在胚胎植入过程中发挥重要的调控作用。联合检测Treg细胞、Tfh17细胞与早期抑制性NK细胞这3种分子标志物，对复发性植入失败女性具有较高的诊断价值，可为辅助生殖技术（ART）中RIF的诊断提供新型潜在生物标志物。本研究结果可为复发性植入失败患者的临床治疗提供参考，也可为未来开展大规模、设计严谨的相关研究提供依据。