Regulation Of Peripheral CD4~+CD25~+Treg In Human Materno-fetal Immunotolerance

A successful pregnancy reflects actually a successful materno-fetal immune tolerance;and pregnancy wastage can be recognized as materno-fetal immune attack. The incidence rate of human spontaneous abortion is as high as 31%in fertile couples. The research on the recurrent spontaneous abortion reveals the 80%unexplained.An increasing number of evidence points that the unexplained recurrent miscarriage is related to the dysfunction of the immune regulation of the pregnant women.Since the etiology of the URM is elusive,treatment of this disease is very difficult.Currently, efficiency and mechanism of the immunotherapy with the lymphocytes isolated from the husband is still not very clear.Hence,to explore an effective treatment is of importance.Recent literature reveals that the CD4~+CD25~+ regulatory T cell expands in the materno-fetal immune regulation during pregnancy.Transcription factor Foxp3 is a gene that regulates the function and development of the CD4~+CD25~+ regulatory T cell. The expression of Foxp3 can influence the function of CD4~+CD25~+ regulatory T cell to some extent.Lots of researches have found that Foxp3 may participate in the immune-related disease.PartⅠExpression features and suppressive ability of Foxp3 in the peripheral Treg cells of women with unexplained recurrent miscarriageObjective:To evaluate the expression of Foxp3 in peripheral blood monocytes of patients with unexplained recurrent miscarriage,and to explore the suppressive effect of the peripheral CD4~+CD25~+ regulatory T cell on.CD4~+CD25~- regulatory T cell proliferation.Methods:The patients with recurrent miscarriage(25) were recruited,and the fertile women with successful delivery(20) as control.The total RNA was isolated from the PBMC,and transcribed into cDNA.The real-time-fluorescence quantitative PCR was used to analyze the transcription of Foxp3.The translation of Foxp3 in CD4~+CD25~+ T cells was analyzed by flow cytometry.The magnetic beads assay was used to isolate the CD4~+CD25~+ T cells and the CD4~+CD25~-T cells.The culture in vitro of the CD4~+CD25~+T cells or CD4~+CD25~-T cells,and the co-culture the CD4~+CD25~+T cells with the CD4~+CD25~-T cells in different effector target ratios.The CCK-8 was used to measure the proliferation and suppressive effects of the CD4~+CD25~+T cells.Results:The expression of Foxp3 mRNA in the PBMC of URM patients was significantly lower than that of the control(P＜0.01).It was showed by flow cytometry that the Foxp3~+ translation in CD4~+CD25~+ T cells of URM was significantly lower than that of the control(P＜0.01).The suppressive effects of the CD4~+CD25~+T cells on CD4~+CD25~-T cell proliferation from the URM patient is significantly lower than that of the control(P＜0.05).Coclusion:The down-regulation of Foxp3 expression and suppressive ability in the peripheral CD4~+CD25~+ regulatory T cells may be involved in URM.PartⅡDydrogesterone induced expansion of the peripheral CD4~+CD25~+Treg improved pregnancy outcome of URMObjective:To investigate the regulatory effects of Dydrogesterone on the CD4+CD25+Treg and pregnancy outcome of URM.Methods:The URM patients were treated orally with dydrogesterone capsule 20mg/day beginning at the 14th day after the BBT rose.We measured the progesterone andβ-HCG in serum at the sixth week and the ninth week,respectively.The proportion of the peripheral CD4~+Foxp3~+T cells was analyzed on the 30th day after beginning the drug,and then the labor was followed up as well.Results:The proportion of the CD4~+Foxp3~+T cells before and after the clinical study of the Dydrogesterone group are 3.4%±1.3%and 4.8%±1.1%respectively,suggesting significant difference compared between the pre-treatment and post-treatment.The values of progesterone at the sixth week and the ninth week in pregnancy after treatment with Dydrogesterone were(29.87±7.01)ng/ml and(22.97±8.79)ng/ml, respectively.The values ofβ-HCG at the sixth and the ninth week in pregnancy after treatment with Dydrogesterone were(37788.5±19112.4) mIU/ml and (106615.3±28880.8) mIU/ml,respectively.The successful labor rates of the administration with Dydrogesterone was 80%.Coclusion:Dydrogesterone could be used to treat URM disease via inducing the peripheral CD4~+CD25~+Treg expansion.