Analysis Of Peripheral Blood Lymphocyte Subsets In Patients With Repeated Implantation Failure

Objective:The purpose of this study was to compare the percentage of peripheral blood lymphocyte subsets in patients with repeated implantation failure(RIF),and to explore the predictive value of related immune indexes in RIF.Methods:A retrospective analysis was made on 161 patients with infertility who were treated in Dalian municipal women and children’s center from September 2018 to December 2020 and tested for lymphocyte subsets.According to the inclusion and exclusion criteria,48 patients with repeated implantation failure were selected as the experimental group.In the same period,113 successful pregnancies after the first cycle of assisted pregnancy were selected as the control group.All subjects were collected from median cubital vein in mid-luteal phase and placed in EDTA anticoagulant vacuum tube.Flow cytometry was used to detect CD3~+T cells,CD3~+CD4~+T cells,CD3~+CD8~+T cells,CD3~+CD4~+/CD3~+CD8~+,CD3~-CD19~+B cells and CD3~-CD16~+/CD56~+NK cells in peripheral blood between the two groups.SPSS 26.0 software was used for statistics and data analysis.It is considered that p<0.05 has statistically significant difference.Results:1.The peripheral blood CD3~+T cells(%)in the control group was 68.80±7.35,and in RIF group was 69.50±8.29,which was not significantly different from the control group(p>0.05).The peripheral blood CD3~+CD4~+T cells(%)in the control group was32.87±6.13,and in the RIF group was 38.09±6.21,which was significantly higher than the control group(p<0.05).The peripheral blood CD3~+CD8~+T cells(%)in the control group was 32.14±7.18,and in the RIF group was 28.17±6.13,which was significantly lower than the control group(p<0.05).The CD3~+CD4~+/CD3~+CD8~+in the control group was 1.10±0.39,and in the RIF group was 1.44±0.41,which was significantly higher than the control group(p<0.05).(reference range:CD3~+T cells 55%-84%,CD3~+CD4~+T cells 31%-60%,CD3~+CD8~+T cells 13%-41%,CD3~+CD4~+/CD3~+CD8~+0.71-2.78)2.The peripheral blood CD3~-CD19~+B cells(%)in the control group was 11.73±4.63,and in the RIF group was 12.05±3.49,which was not significantly different from the control group(p>0.05).(reference range:CD3~-CD19~+B cells 5%-20%)3.The peripheral blood CD3~-CD16~+/CD56~+NK cells(%)in the control group was18.92±7.72,and in the RIF group was 17.47±9.07,which was not significantly different from the control group(p>0.05).(reference range:CD3~-CD16~+/CD56~+NK cells7%-36%)4.Among the RIF patients,there was significant difference in peripheral blood CD3~+CD4~+/CD3~+CD8~+among the patients with 2,3 and≥4 times of transplantation failure(p<0.05),and there was significant difference between 2 times(1.24±0.49)and≥4 times(1.94±0.15)(p<0.05).However,there was no significant difference in peripheral blood CD3~+T cell,CD3~+CD4~+T cell,CD3~+CD4~+T cell,B cell and NK cell among patients with different times of transplantation failure(p>0.05).5.ROC curve analysis showed that the percentage of peripheral blood CD3~+CD4~+T cells was>35.6%(AUC=0.713),with a specificity of 65.5%and a sensitivity of 68.8%for RIF;the percentage of peripheral blood CD3~+CD8~+T cells was<31.55%(AUC=0.659),with a specificity of 75.0%and a sensitivity of 54.0%for RIF;peripheral blood CD3~+CD4~+/CD3~+CD8~+>1.155(AUC=0.747),with a specificity of 66.4%and a sensitivity of 78.1%for RIF.Conelusions:1.In the peripheral blood of patients with RIF,CD3~+CD4~+T cells(%)increased,CD3~+CD8~+T cells(%)decreased and CD3~+CD4~+/CD3~+CD8~+increased,suggesting that the imbalance between CD3~+CD4~+T cells and CD3~+CD8~+T cells may may be one of the factors affecting embryo implantation.2.Peripheral blood CD3~+CD4~+T cell percentage greater than 35.6%,CD3~+CD8~+T cell percentage less than 31.55%and CD3~+CD4~+/CD3~+CD8~+greater than 1.155 can be used as criteria for predicting T lymphocyte subsets in patients with unexplained repeated implantation failure.