Dii ESA intraperitoneally at G5 (G5 ip), with the abortion rate of nearly 100 . Some embryos and placentas exhibited a necrotic and haemorrhagic appearance after the administration of T. Title Loaded From File gondii ESA intraperitoneally at G10 (G10 ip), with the abortion rate up to 56.20 . However, after the injection of T. gondii ESA at G15, there was no visible fetal abnormality in pregnant mice, which was consistent with the mice in control group (Figure 1).Injection of T. gondii ESA at the Early and Intermediate Stages of Pregnancy Reduces the Frequency and Function of CD4+CD25+Foxp3+ T Cells of MiceIt has been previously determined that T. gondii has the ability to diminish the number of CD4+CD25+Foxp3+ T cells of mice during the gestation [17]. Consistent with those data, we found that the administration of T. gondii ESA at early (G5) and intermediate (G10) stages of pregnancy could also lead to the decrease of CD4+CD25+Foxp3+ T cells. However, after the injection of T. gondii ESA at the late pregnancy (G15), the percentage of CD4+CD25+Foxp3+ T cells increased compared with that of the control group (Figure 2A). The phenomenon could also be observed in the inguinal lymph nodes (LN) and peripheral blood lymphocytes (PBL) (Figure 2B and 2C), suggesting that T. gondii ESA induced global changes of CD4+CD25+Foxp3+ T cells. Next, we tested whether the regulatory function of these cells from the injected group ofT. gondii ESA Induced Tregs Dysfunctionmice had been damaged by evaluating the suppressing proliferation of CD4+CD25+ T cells in vitro and Th2/Th1-like responses in vivo. We obtained purified CD4+CD25+ T cells from the normal pregnant mice and the mice with T. gondii ESA-injection at G5, G10 and G15, 1315463 respectively. The decreased suppressive ability of CD4+CD25+ T cells was observed in mice with the ESA-injection at G5 and G10. However, the inhibitory capacity of the CD4+CD25+ T cells was enhanced after the injection of T. gondii ESA at G15 (Figure 2D). Due to the capacity of CD4+CD25+ Treg cells controlling potentially detrimental IFN-c Title Loaded From File reactions during pregnancy [28], we detected the serum level of IFN-c after the injection of T. gondii ESA. We found that the serum level of IFN-c was up to 448.3 pg/ml at G5 ip, suggesting that the activity of CD4+CD25+ Tregs on the suppression of IFN-c production was impaired (Figure 2E). As expected, in all groups of mice, the serum IL-4 levels were not obviously affected (Figure 2F). Taken together, the results showed that the frequency and function of CD4+CD25+Foxp3+ T cells were diminished after the injection of T. gondii ESA at early and intermediate stages of pregnancy.Injection of T. gondii ESA at the Intermediate Stage of Pregnancy Decreases the Levels of Foxp3 mRNA and Protein at the Maternal-fetal Interface of MiceA complex regulation of immune response at the maternal-fetal interface promotes tolerance of paternally derived antigens [29]. To determine if the reduction of CD4+CD25+ Tregs also occurred at the maternal-fetal interface, we analyzed the expression levels of Foxp3 mRNA and protein in the placentas of mice with T. gondii ESA-injection at G10 and G15. The results showed that the expression levels of placental Foxp3 mRNA and protein were decreased at G10, but increased at G15, as compared with the control groups (Figure 3A and 3B). The distribution of Foxp3+ cells at the maternal-fetal interfaces was also observed by immunohistochemistry. As shown in Figure 3C and 3D, the placentas of mice with T.Dii ESA intraperitoneally at G5 (G5 ip), with the abortion rate of nearly 100 . Some embryos and placentas exhibited a necrotic and haemorrhagic appearance after the administration of T. gondii ESA intraperitoneally at G10 (G10 ip), with the abortion rate up to 56.20 . However, after the injection of T. gondii ESA at G15, there was no visible fetal abnormality in pregnant mice, which was consistent with the mice in control group (Figure 1).Injection of T. gondii ESA at the Early and Intermediate Stages of Pregnancy Reduces the Frequency and Function of CD4+CD25+Foxp3+ T Cells of MiceIt has been previously determined that T. gondii has the ability to diminish the number of CD4+CD25+Foxp3+ T cells of mice during the gestation [17]. Consistent with those data, we found that the administration of T. gondii ESA at early (G5) and intermediate (G10) stages of pregnancy could also lead to the decrease of CD4+CD25+Foxp3+ T cells. However, after the injection of T. gondii ESA at the late pregnancy (G15), the percentage of CD4+CD25+Foxp3+ T cells increased compared with that of the control group (Figure 2A). The phenomenon could also be observed in the inguinal lymph nodes (LN) and peripheral blood lymphocytes (PBL) (Figure 2B and 2C), suggesting that T. gondii ESA induced global changes of CD4+CD25+Foxp3+ T cells. Next, we tested whether the regulatory function of these cells from the injected group ofT. gondii ESA Induced Tregs Dysfunctionmice had been damaged by evaluating the suppressing proliferation of CD4+CD25+ T cells in vitro and Th2/Th1-like responses in vivo. We obtained purified CD4+CD25+ T cells from the normal pregnant mice and the mice with T. gondii ESA-injection at G5, G10 and G15, 1315463 respectively. The decreased suppressive ability of CD4+CD25+ T cells was observed in mice with the ESA-injection at G5 and G10. However, the inhibitory capacity of the CD4+CD25+ T cells was enhanced after the injection of T. gondii ESA at G15 (Figure 2D). Due to the capacity of CD4+CD25+ Treg cells controlling potentially detrimental IFN-c reactions during pregnancy [28], we detected the serum level of IFN-c after the injection of T. gondii ESA. We found that the serum level of IFN-c was up to 448.3 pg/ml at G5 ip, suggesting that the activity of CD4+CD25+ Tregs on the suppression of IFN-c production was impaired (Figure 2E). As expected, in all groups of mice, the serum IL-4 levels were not obviously affected (Figure 2F). Taken together, the results showed that the frequency and function of CD4+CD25+Foxp3+ T cells were diminished after the injection of T. gondii ESA at early and intermediate stages of pregnancy.Injection of T. gondii ESA at the Intermediate Stage of Pregnancy Decreases the Levels of Foxp3 mRNA and Protein at the Maternal-fetal Interface of MiceA complex regulation of immune response at the maternal-fetal interface promotes tolerance of paternally derived antigens [29]. To determine if the reduction of CD4+CD25+ Tregs also occurred at the maternal-fetal interface, we analyzed the expression levels of Foxp3 mRNA and protein in the placentas of mice with T. gondii ESA-injection at G10 and G15. The results showed that the expression levels of placental Foxp3 mRNA and protein were decreased at G10, but increased at G15, as compared with the control groups (Figure 3A and 3B). The distribution of Foxp3+ cells at the maternal-fetal interfaces was also observed by immunohistochemistry. As shown in Figure 3C and 3D, the placentas of mice with T.
