Oikonomou Georgia, Vlachadis Nikolaos, Tsamadias Vassilios, Lambrinoudaki Irene, Deligeoroglou Efthymios, Vlahos Nikolaos F, Economou Emmanuel
Second Department of Obstetrics and Gynecology, National and Kapodistrian University of Athens Medical School, Aretaieio University Hospital, Athens, GRC.
Department of Obstetrics and Gynecology, General Hospital of Messinia, Kalamata, GRC.
Cureus. 2023 Mar 23;15(3):e36584. doi: 10.7759/cureus.36584. eCollection 2023 Mar.
The maternal immune system has a major role in the successful embryo implantation and maintenance of the pregnancy. This study aimed to investigate the maternal immunophenotyping profile (percentage of Natural Killer [NK] cells and the CD4/CD8 [cluster designation] ratio in peripheral blood lymphocytes) and the HLA (Human Leukocyte Antigen)-DQA1 alleles sharing in infertile couples.
This cross-sectional study included 78 women who had experienced at least two spontaneous miscarriages and 110 women with a history of recurrent implantation failures after in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) and embryo transfer (ET) (IVF-ET failures). The NK cell percentage and the CD4/CD8 ratio were determined by flow cytometry. Genotyping of the HLA-DQA1 alleles was carried out for all women and their partners, and couple HLA-DQA1 compatibility was expressed as the percentage of common HLA-DQA1 alleles (totaling 35 alleles) shared between spouses to the sum of the unique alleles observed.
In women with recurrent miscarriages, high values (%) of the NK population with a median (interquartile range [IQR]) of 10.3% (7.7% to 12.5%) and CD4/CD8 ratio (1.7) (1.5 to 2.1) were found. In women with IVF-ET failures, the (%) NK population (10.5%) (8.6% to 12.5%) and CD4/CD8 ratio (1.8) (1.5 to 2.1) were similarly increased (p=0.390, and p=0.490, respectively). The proportion of women with >10% NK cells was 53.8% and 58.2% in women with miscarriages and IVF-ET failures, respectively (p=0.554). The prevalence of HLA-DQA15 allele carriage was elevated in women with miscarriages as well as those with IVF-ET failures (52.6% and 61.8%, respectively; p=0.206). The proportion of couples with high (>50%) HLA-DQA1 sharing was 65.4% in the group with miscarriages and 73.6% in the group with IVF-ET failures, respectively (p=0.222). The CD4/CD8 ratio was statistically significantly positively correlated with the (%) NK population in women with IVF-ET failures (rho = 0.297, p=0.002) and with the (%) HLA-DQA1 sharing in the group with miscarriages (rho = 0.266, p=0.019). The couples in which both spouses were carriers of the HLA-DQA15 allele had an increased probability of high (>50%) HLA-DQA1 compatibility compared with the couples in which neither of the spouses carried the allele in the miscarriage group (OR = 24.3, 95% CI: 3.0 to 198.9, p<0.001), and the IVF-ET failure group (OR = 10.5, 95% CI: 2.2 to 49.8, p<0.001).
The peripheral NK (%) population and CD4/CD8 ratio, as well as the prevalence of the HLA-DQA15 allele, were elevated in women with recurrent miscarriages and IVF-ET failures. Furthermore, these couples with negative reproductive outcomes had a high percentage of HLA-DQA1 allele similarity. The presence of the HLA-DQA15 allele in spouses was strongly associated with overall couple HLA-DQA1 compatibility, implying that it could be used as a surrogate marker for assessing overall immunological compatibility in infertile couples.
母体免疫系统在胚胎成功着床及维持妊娠过程中发挥着重要作用。本研究旨在调查不孕夫妇的母体免疫表型特征（外周血淋巴细胞中自然杀伤[NK]细胞百分比及CD4/CD8[簇分化]比值）以及人类白细胞抗原（HLA）-DQA1等位基因共享情况。
这项横断面研究纳入了78名经历过至少两次自然流产的女性以及110名在体外受精（IVF）或卵胞浆内单精子注射（ICSI）及胚胎移植（ET）后有反复着床失败史（IVF-ET失败）的女性。通过流式细胞术测定NK细胞百分比及CD4/CD8比值。对所有女性及其配偶进行HLA-DQA1等位基因基因分型，夫妇HLA-DQA1相容性以配偶间共享的常见HLA-DQA1等位基因（共35个等位基因）占所观察到的独特等位基因总和的百分比来表示。
在反复流产的女性中，发现NK细胞群体百分比中位数（四分位间距[IQR]）较高，为10.3%（7.7%至12.5%），CD4/CD8比值为1.7（1.5至2.1）。在IVF-ET失败的女性中，NK细胞群体百分比（10.5%）（8.6%至12.5%）及CD4/CD8比值（1.8）（1.5至2.1）同样升高（分别为p = 0.390及p = 0.490）。NK细胞>10%的女性比例在反复流产女性及IVF-ET失败女性中分别为53.8%和58.2%（p = 0.554）。HLA-DQA15等位基因携带率在反复流产女性及IVF-ET失败女性中均升高（分别为52.6%和61.8%；p = 0.206）。HLA-DQA1共享率高（>50%）的夫妇比例在反复流产组为65.4%，在IVF-ET失败组为73.6%（p = 0.222）。在IVF-ET失败的女性中，CD4/CD8比值与NK细胞群体百分比呈显著正相关（rho = 0.297，p = 0.002），在反复流产组中与HLA-DQA1共享率呈正相关（rho = 0.266，p = 0.019）。与配偶双方均不携带该等位基因的夫妇相比，在反复流产组中，配偶双方均为HLA-DQA15等位基因携带者的夫妇HLA-DQA1相容性高（>50%）概率增加（比值比[OR]=24.3，95%置信区间[CI]：3.0至198.9，p<0.001），在IVF-ET失败组中为（OR = 10.5，95%CI：2.2至49.8，p<0.001）。
反复流产及IVF-ET失败女性的外周NK细胞百分比及CD4/CD8比值，以及HLA-DQA15等位基因携带率均升高。此外，这些生殖结局不良的夫妇HLA-DQA1等位基因相似性较高。配偶中存在HLA-DQA15等位基因与夫妇总体HLA-DQA1相容性密切相关，这意味着它可作为评估不孕夫妇总体免疫相容性的替代标志物。
