Sims IVF Clinic, Clonskeagh Road, Clonskeagh, Dublin 14, Ireland.
RCSI, Beaumont Hospital, Dublin, Ireland.
J Assist Reprod Genet. 2019 May;36(5):837-846. doi: 10.1007/s10815-019-01427-8. Epub 2019 Mar 7.
The uterine immunophenotype is relatively poorly understood, with most studies reporting proportions/percentages. A novel technique to calculate local endometrial lymphocyte concentrations is described, and used to compare results between aetiological subgroups such as repeated implantation failure (RIF) and recurrent pregnancy loss (RPL) with male-factor controls.
455 patients had an endometrial biopsy performed. Background history on initial presentation was used to subdivide the population into RIF (n = 149), RPL (n = 121), primary (n = 76) and secondary infertility (n = 80). A control group was identified comprising male factor infertility aetiology with all female investigations normal (n = 29). Endometrial Tissue was assessed using a comprehensive multi-parameter panel. Lymphocyte subpopulations were calculated using flowcount flurospheres and a mathematical correction applied to determine concentrations per milligram of tissue, based on original biopsy weight and volumetric dilutions.
The flow cytometry technique was successful in determining population centiles for concentrations of endometrial lymphocyte subsets. Distinct differences were noted across the patient groups. Th2 concentrations were significantly higher in the controls (p = 0.0002). All RPL/infertile populations had increased concentrations of peripheral type NK's (p = 0.016) and B cells (p = 0.045). Relative to male factor controls, CD4+ and CD8+ T lymphocyte populations were increased in RPL patients, and reduced in those with a history of RIF. Th1 concentrations were elevated in the adverse outcome groups (p = 0.032). Concentration centiles alone do not appear to accurately predict outcome with subsequent treatment.
Endometrial biopsy analysis by flow cytometry can provide detailed analysis of constituent lymphocyte subsets by concentration as well as proportion. This novel approach provides additional independent data to further assess the significance of endometrial changes in the setting of reproductive failure.
子宫免疫表型的了解相对较少，大多数研究报告的是比例/百分比。本文描述了一种计算局部子宫内膜淋巴细胞浓度的新方法，并将其用于比较反复着床失败（RIF）和复发性妊娠丢失（RPL）等病因亚组与男性因素对照组之间的结果。
对 455 名患者进行了子宫内膜活检。根据初始表现的背景病史，将人群细分为 RIF（n=149）、RPL（n=121）、原发性（n=76）和继发性不孕（n=80）。对照组为男性因素不孕病因，所有女性检查均正常（n=29）。使用综合多参数面板评估子宫内膜组织。使用流式细胞术荧光球计算淋巴细胞亚群，并应用数学校正方法根据原始活检重量和体积稀释度，计算每毫克组织的浓度。
流式细胞术技术成功地确定了子宫内膜淋巴细胞亚群浓度的百分位数。在不同的患者组中观察到明显的差异。对照组 Th2 浓度显著升高（p=0.0002）。所有 RPL/不孕人群的外周型 NK 细胞（p=0.016）和 B 细胞（p=0.045）浓度均升高。与男性因素对照组相比，RPL 患者的 CD4+和 CD8+T 淋巴细胞群增加，而 RIF 病史患者的减少。Th1 浓度在不良结局组中升高（p=0.032）。浓度百分位数本身似乎不能准确预测随后治疗的结果。
通过流式细胞术对子宫内膜活检进行分析，可以提供细胞浓度和比例的详细分析。这种新方法提供了额外的独立数据，以进一步评估生殖失败背景下子宫内膜变化的意义。
