Study On The Relationship Between Expression Of CD4+ CD25+ Regulatory T Cells And Th1 And Th2 In Unexplained Recurrent Spontaneous Abortion

Chapter I Clinical Study on Relationship between unexplained recurrentspontaneous abortion and Expression of CD4+CD25+ Regulatory T Cells and Th1 and Th2 in immune recurrent spontaneous abortionObjective: To investigate the relationship between unexplained recurrentspontaneous abortion and the expression of CD4+CD25+regulatory T cells and Th1 and Th2 in clinical.Methods: Forty-five cases of clinical study subjects were randomly selected according to the criteria for immune recurrent spontaneous abortion as an abortion group,normal pregnant women as the control group.Peripheral blood of the subjects was collected,and the proportion of CD4+CD25+ regulatory T cells was measured by flow cytometry,and Th1 cytokines(IFN-γ,IL-2)and Th2 cytokines(IL-4,IL)were detected by ELISA.The expression of Foxp3 m RNA was detected by RT-PCR.Result: Flow cytometry results showed that the proportion of CD4+CD25+ regulatory T in the control group was approximately(7.95 ± 1.13)%,and the proportion of CD4+CD25+ regulatory T in the abortion group was approximately(4.06 ± 1.53)%.There was a significant difference between the two groups.The results of ELISA showed that the levels of IFN-γ and IL-2 in the abortion group were significantly higher than those in the control group,and the levels of IL-4 and IL-10 were significantly lower.Statistical analysis of RT-PCR results showed that the Foxp3 m RNA level in the abortion group was significantly lower than that in the control group,with significant differences between the groups.Conclusion: In Clinical,the proportion of CD4+CD25+ regulatory T in URSA patients decreases,and the ratio of Th1/Th2 increases,which may be one of the monitoring indicators of URSA patients.Chapter 2 Evaluation of clinical effects of active immunization combined with progesterone treatment for unexplained recurrentspontaneous abortionObjective: To investigate the effect of progesterone combined with active immunotherapy on the expression of CD4+CD25+ regulatory T cells and Th1 and Th2 in URSA patients.Mehods: Fifty-two patients received progestogen combined with active immunotherapy as the treatment group and the remaining 45 patients were used as abortion group.The patients in the treatment group were treated with active immunotherapy and the progesterone was administered during the pregnancy after the blocking antibody BA was converted to positive.The abortion group did not undergo any of the above treatments.Peripheral blood was collected from the subjects,flow cytometry was used to measure the proportion of CD4+CD25+ regulatory T cells,RT-PCR was used to detect the expression of Foxp3 m RNA,and ELISA was used to detect cell cytokines levels;serum progesterone levels detected by chemiluminescence;patient follow-up statistics pregnancy outcomes.Results: Flow cytometry results showed that the proportion of CD4+CD25+ regulatory T in the abortion group was lower than the treatment group.Statistical analysis of RT-PCR results showed that the Foxp3 m RNA level in the treatment group was significantly higher than that in the abortive group.The results of ELISA showed that compared with the abortion group,the levels of IFN-γ and IL-2 in the treatment group were significantly lower,and the levels of IL-4 and IL-10 were significantly higher.After progesterone combined with active immunotherapy,serum progesterone levels were significantly higher in the 8 weeks of pregnancy and 10 weeks of pregnancy than in the control group.The success rate of pregnancy in the treatment group was 86.54%,which was significantly higher than that in the abortion group(66.67%).The successful delivery rate was 95.55% in the treatment group,which was significantly higher than that in the abortion group(46.67%).Conclusion: Progesterone combined with active immunotherapy is an effective method for the treatment of URSA.The ratio of CD4+CD25+ regulatory T and Th1/Th2 may be used as one of the monitoring indicators for URSA patients.Chapter 3 The Expression of CD4+CD25+ Regulatory T Cells and the Relationship between Th1 and Th2 in unexplained recurrentspontaneous abortion Mice ModelObjective: To investigate the effect of progesterone combined with active immunotherapy on the expression of CD4+CD25+ regulatory T cells and Th1 and Th2 in a mouse model of unexplained recurrentspontaneous abortion.Methods: CBA/JXDBA/2J and CBA/J x BALB/c were selected for mating to prepare abortive and control mouse models.The mice in the treatment group were treated with active immunotherapy and progesterone was administered during pregnancy.The abortion group did not undergo any of the above treatments.Flow cytometry,RT-PCR and Western blot methods were used as the description in above.HE staining was used to observe the histological changes of the endometrium.Immunohistochemistry was used to detect PGR and Foxp3 protein expression in the uterus.Result: Compared with the control group,the proportion of CD4+CD25+ regulatory T in the abortion group was lower and higher in treatment group in both spleen and blood.Western blot results showed that compared with the control group,the Foxp3 protein in the abortion group was significantly decreased.Compared with the abortive group,the Foxp3 protein in the treated group was significantly higher than that in the control group.RT-PCR results showed that compared with the control group,the Foxp3 m RNA in the abortion group was significantly decreased.Compared with the abortion group,the Foxp3 m RNA in the treated group was significantly increased.ELISA results showed that compared with the abortion group,the levels of IFN-γ and IL-2 in the treatment group were significantly lower,and the levels of IL-4 and IL-10 were significantly higher.The results of HE staining showed that compared with the control group,the thickness of the endometrium of the mice in the abortion group decreased,and the inflammatory cells infiltrated,and the number of glands decreased.The endometrium thickness of the treated mice increased,the infiltration of inflammatory cells weakened,and the number of glands increased.HE results showed that the recurrent abortion mice had severe inflammatory reactions in the endometrium,insufficient supply of progesterone,and reduced nutrient supply required for embryonic growth.After progesterone combined with active immunotherapy,the inflammatory response in mice was reduced and progesterone and nutrient supply were improved.The uterus of the control group showed strong PGR protein expression.The expression level of PGR protein in the aborted mice was significantly decreased.The expression of PGR protein in the uterus of the treated group was increased.Foxp3 protein immunohistochemistry results showed that the control group mouse uterus showed strong Foxp3 protein expression,abortion group mouse PGR protein expression was significantly reduced,the expression of PGR protein in the uterus of the treatment group increased.Conclusion: Progesterone combined with active immunotherapy can significantly improve the immune status of URSA mice and can be used as an effective method to treat URSA in mice.