Research On Th17 And Foxp3~+ Regulatory T Cells Imbalance And Expression Of Related Cytokines In Patients With Repeated Implantation Failure After Ivf

PARTⅠExpression of the T-regulatory cell transcription factor Foxp3 and Th17-related cytokine IL-17A in endometrial tissueObjective:This study aim to evaluate the expression of the T-regulatory cell transcription factor Foxp3 and Th17-related cytokine IL-17A in endometrial tissue of patients with with repeated implantation failure after IVF,and discuss the relationship between Th17/Treg cells and RIF.Methods:Endometrial biopsies were collected during the mid-secretory phase of the menstrual cycle(days LH₊₆to LH₊₇)from patients with repeated implantation failure after IVF（RIF,n=26）,or women classified as proven fertile（control,n=27）.Expression of the T-regulatory cell transcription factor Foxp3 and Th17-related cytokine IL-17A in endometrial tissue were detected by immunocytochemistry.Results:The density of Foxp3⁺cells in endometrium from women with repeated implatation failure after IVF was significantly lower than in endometrium from control women（51.0±17.6%vs 65.6±16.7%,P<0.05）.The density of IL-17A⁺cells in endometrium from women with repeated implatation failure after IVF was significantly higher than in endometrium from control women（57.7±18.4%vs 48.9±12.8%,P<0.05）.Conclusions:Decreased Foxp3⁺cells and increased IL-17A⁺cells may affect on uterine receptivity during the mid-luteal phase.Imbalance between Th17 and Foxp⁺Treg cells may correlated with pregnancy outcome of RIF.PART ⅡImbalance between Th17 and Foxp3+regulatory T cells in peripheral blood of patients with repeated implantation failureObjective: The purpose of this study was to determine whether there is an imbalance in IL-17+ T（Th17）and Foxp3+ regulatory T（Treg）cells in peripheral blood of patients with repeated implantation failure（RIF）.Methods: Flow cytometric analysis was performed to evaluated the peripheral percentages of Th17 cells and Foxp3+Treg cells within the CD4+ Tcell pool in women with RIF（n=25）and fertile non-pregnant controls（n=27）at the late proliferative phase of menstrual cycle.At the same time,the secretion of the Th17/Treg-linked cytokines were assessed by enzymelinkedimmunosorbent assay（ELISA）and m RNA expression of transcription factors were detected by quantitative real-time PCR（q RT-PCR）.Results: The peripheral percentage of Th17 cells and Th17/Treg ratio was significantly higher at the late proliferative phase in patients with RIF（1.50±0.38%;median）than controls（1.16±0.48%;median）（all P<0.05）,whereas the peripheral percentage of Foxp3+ Treg cells was lower in RIF women than fertile women（2.43±0.97% vs 3.24±0.82%,P < 0.05）.The ratio of Th17/Treg was higher in the RIF group than the control group（median 0.58 vs0.34;P <0.05）.The percentage of Foxp3+ Treg cells conversely correlates with that of Th17 cells in RIF or NNP group（r =-0.414;r =-0.397,all P<0.05）.The secretion of the Treg-related cytokines TGF-beta and IL-10 were decreased（0.26±0.20 vs0.63±0.24;25.30±1.74 vs 26.72±2.68;all P < 0.05）and the secretion of the Th17-related cytokines IL-6,IL-17 A and IL-23 were increased（13.64±9.17 vs8.77±3.26;median 23.83 vs 20.21;median 7.98 vs 7.79;all P<0.05）in serum in RIF women,compared with fertile non-pregnant women.The ratios of Treg/Th17 cytokines were significantly lower in RIF group than in control group（P< 0.05）.The expression of the Treg-specific transcription factor Foxp3（0.87±0.54 vs 1.27±0.73,P<0.05）was decreased and the expression of Th17-related transcription factor retinoic acid-related orphan receptor γt（RORγt）（1.53±0.74 vs 1.15±0.50,P<0.05）was increased in peripheral blood in women with RIF,compared with controls.Conclusions: There may exist an imbalance of Th17/Treg in peripheral blood of patients with repeated implantation failure（RIF）.Th17/Treg balance may be a new direction for treatment of RIF.PART ⅢAlteration of Th17 and Treg cells in patients with repeated implantation failure before and after prednisone administrationObjective : The purpose of this study was to determine whether corticoid therapy could be use as an effective treatment for those patients with repeated implantation failure who have an imbalance between regulatory T and Th17 cells.Methods : Flow cytometric analysis was performed to evaluated the peripheral percentages of CD4+ CD25+ Foxp3+ Treg cells and CD4+ IL-17+ T cells with CD4+ T-cell population in women with RIF（n=25）before and after prednisone administration.At the same time,the secretion of the Th17/Tregassociated cytokines were assessed by enzyme-linked immunosorbent assay（ELISA）and m RNA expression of transcription factors were detected by quantitative real-time PCR（q RT-PCR）.Results : The peripheral percentage of Th17 cells was significantly lower（median 1.18% vs 1.39%,P<0.05）and the percentage of Foxp3+ Treg cells was significantly higher（median 2.88% vs 2.36%,P < 0.05）at the late proliferative phase in patients given prednisone.Prednisone administration may induce a decrease in the Th17/Treg ratio and increase regulatory T cells level which may be beneficial for the maintenance of pregnancy（median 0.54 vs 0.42,P<0.05）.The secretion of the Treg-related cytokines TGF-beta and IL-10 were increased in serum（median 0.53 vs 0.09;median 26.24 vs 26.09;all P<0.05）,while the secretion of the Th17-related cytokines IL-6,IL-17 A and IL-23 were decreased after prednisone administration（median 7.38 vs 12.59;median 23.11 vs 24.94;median 7.75 vs 7.93;all P<0.05）.The ratios of Treg/Th17 cytokines were significantly higher after prednisone administration as compared with before prednisone administration（P<0.05）.The expression of the Treg-specific transcription factor Foxp3 increased and the expression of Th17 related transcription factor retinoic acid-related orphan receptor γ t（ROR γ t）decreased in peripheral blood as measured by q RT-PCR（median 0.85 vs 0.54;median 0.83 vs 1.60;all P <0.05）.The clinical pregnancy rate of FET in the19 patients was 29.3%（5/19）after prednisone administration.Conclusions : Corticoid therapy was able to affect Th17/Treg balance and increase the percentage of Treg cells in peripheral blood lymphocytes which are beneficial to pregnancy.