外周血淋巴细胞亚群与不明原因反复种植失败的相关性研究

Study on Correlation between Peripheral Blood Lymphocyte Subsets and Unexplained Repeated Implantation Failure

目的:分析外周血淋巴细胞亚群与不明原因反复种植失败(URIF)的关系,探讨其对URIF诊断的预测价值。方法:选取2019年10月1日至2021年12月1日在昆明医科大学第二附属医院生殖医学科就诊的156例URIF患者为URIF组,同时匹配年龄,纳入近一年有健康分娩史的41例女性作为对照组,通过流式细胞技术完成两组受试者外周血淋巴细胞亚群百分比的检测,对检测结果进行统计学分析,并采用ROC曲线评估淋巴细胞亚群对URIF的预测效果。结果:与对照组相比,URIF组CD3^(+)CD4^(+)[(34.03±7.26)%vs.(36.79±6.35)%,P=0.017]、CD3^(+)HLA-DR^(+)[(2.60±2.28)%vs.(3.60±2.39)%,P=0.017]、CD3^(+)CD16^(+)CD56^(+)[(1.24±1.04)%vs.(2.62±2.57)%,P<0.000]及CD4^(+)/CD8^(+)(1.37±0.48 vs.1.57±0.51,P=0.023)显著升高,且在一定范围内随胚胎种植失败次数增加而增高。ROC分析示,CD3^(+)CD4^(+)>35.35%(AUC 0.624)、CD3^(+)HLA-DR^(+)>2.35%(AUC 0.669)、CD3^(+)CD16^(+)CD56^(+)>1.86%(AUC 0.660)和CD4^(+)/CD8^(+)>1.26(AUC 0.628)可单独作为预判URIF免疫异常的预测指标,两两联合指标中,CD3^(+)HLA-DR^(+)与CD3^(+)CD16^(+)CD56^(+)联合预测准确度最高(AUC 0.739,敏感度73.3%,特异度68.3%),四者联合后预测价值进一步增高(AUC 0.767,敏感度68.6%,特异度73.2%)。结论:URIF患者体内存在免疫功能紊乱,其外周血淋巴细胞亚群的失衡可能是导致胚胎种植失败的重要因素。CD4^(+)/CD8^(+)、CD3^(+)CD4^(+)、CD3^(+)HLA-DR^(+)和CD3^(+)CD16^(+)CD56^(+)可作为判断URIF患者外周血淋巴细胞亚群异常的指标,各指标联合后能提高预测的准确性,对辅助生殖助孕过程中URIF人群的动态监测和早期干预有积极的参考意义。

Objective:To analyze the correlation between peripheral blood lymphocyte subsets and unexplained repeated implantation failure(URIF),and to investigate its predictive value for the diagnosis of URIF.Methods:A total of 156 patients with URIF who underwent treatment in the Department of Reproductive Medicine,The Second Affiliated Hospital of Kunming Medical University from October 1,2019 to December 1,2021 was selected as the URIF group.Meanwhile,age-matched 41 women with a history of healthy delivery in the past one year were selected as the control group.The percentages of peripheral blood lymphocyte subsets in the two groups were measured by flow cytometry,and the results were statistically analyzed.Receiver operating characteristic(ROC)curve was used to evaluate the predictive effect of lymphocyte subsets on URIF.Results:Compared with the control group,the percentage of CD3^(+)CD4^(+)[(34.03±7.26)%vs.(36.79±6.35)%,P=0.017]、CD3^(+)HLA-DR^(+)[(2.60±2.28)%vs.(3.60±2.39)%,P=0.017]、CD3^(+)CD16^(+)CD56^(+)[(1.24±1.04)%vs.(2.62±2.57)%,P<0.000]and CD4^(+)/CD8^(+)(1.37±0.48 vs.1.57±0.51,P=0.023)were significantly increased in URIF group,and were related to the increase of previous failure times to a certain extent.ROC analysis showed that CD3^(+)CD4^(+)>35.35%(AUC 0.624),CD3^(+)HLA-DR^(+)>2.35%(AUC 0.669),CD3^(+)CD16^(+)CD56^(+)>1.86%(AUC 0.660)and CD4^(+)/CD8^(+)>1.26(AUC 0.628)could be used as an independent predictor for the diagnosis of URIF.Among the pair-wise combined indexes,CD3^(+)HLA-DR^(+)combined with CD3^(+)CD16^(+)CD56^(+)had the highest prediction accuracy(AUC 0.739,Sensitivity 73.3%,Specificity 68.3%).The combination of the four indicators had the highest accuracy(AUC 0.767,Sensitivity 68.6%,Specificity 73.2%).Conclusions:There is immune dysfunction in patients with URIF,and the imbalance of peripheral blood lymphocyte subsets may be an important factor leading to embryo implantation failure.CD3^(+)CD4^(+)、CD4^(+)/CD8^(+)、CD3^(+)HLA-DR^(+)and CD3^(+)CD16^(+)CD56^(+)could be used as independent indicators for the diagnosis of abnormal peripheral blood lymphocyte subsets in patients with URIF.The combination of them improves the accuracy of prediction,and it has a positive reference significance for dynamic monitoring and early intervention of URIF patients in the process of assisted reproduction technology.