The Expression Of HLA-G And IL-10, Molecules Related With Immunotolerance In Adenomyosis

PARTâ… Expression of HLA-G in Patients with AdenomyosisObjective:To demonstrate the expression of HLA-G,an immunotolerance related molecule,in the eutopic and ectopic endometrium of patients with adenomyosis;to compare the different levels of HLA-G in endometrium between normal women and patients with adenomyosis.To illustrate the role of HLA-G in establishing the state of immunotolerance and the possible contribution of HLA-G to understanding the pathogenesis of adenomyosis.Methods:The surgical biopses from 34 patients with adenomyosis were collected as experimental group,30 biopses normal endometrium collected as control groups.Immunohistological staining and hybridization in situ were used to detect the protein and mRNA of HLA-G in normal endometrium,eutopic and ectopic endometrium of adenomyosis.Quantitative analysis of HLA-G expression was performed according to H score system.The software of SPSS13.0 was used to analyze the experimental data.Results:1.In all the normal endometrial samples,neither epithelial nor stromal HLA-G expression was observed throughout the menstrual cycle.2.Epithelial and stromal HLA-G immunoreactivity was observed in all eutopic endometrium and adenomyosis foci of women with adenomyosis in both proliferative and secretory phases.Similar to the normal endometrial samples,the epithelial cells of the eutopic endometrium and adenomyosis foci showed higher staining intensity than the stromal cells.The epithelial and stromal staining was mostly cytoplasmic and not in nuclear. In the eutopic endometrium of women with adenomyosis,both proliferative and secretory epithelial cells showed HLA-G positive and no cyclic variance was found with no statistical difference between the two phases(P>0.05).The Hscore for the proliferative and secretory stomal cells was not found statistically different(P>0.05).The ectopic epithelial cells of adenomyosis have statistically higher Hscore than the eutopic epithelial cells(P<0.05);however,the ectopic stromal cells have lower Hscore than the eutopic stromal cells(P<0.05).3.A good correlationship was found between HLA-G expression in ectopic and eutopic endometrium in patients with adenomyosis,both in glands(r=0.89,P<0.0001)and in stroma(r=0.78,P<0.0001).4. The positive rate for HLA-G mRNA in normal endometrium,eutopic endometrium and ectopic endometrium is 1/30,28/34,31/34,respectively.The positive signal mostly localized in epithelial cells and stromal cells have weak signal.Conclusions:1.HLA-GmRNA and HLA-G protein are found in the eutopic and ectopic endometrium of patients with adenomyosis,whereas not found in normal endometrium.These findings indicate that HLA-G may involve in the pathogenesis of adenomyosis by establishing the local microenviroment of immunotolerance and immunosuppression.2.The good correlationship between HLA-G expression in eutopic and ectopic endometrium in patients with adenomyosis indicates that the characterism of eutopic endometrium may play an important role in the pathogenesis of adenomyosis. PARTâ…¡Expression of IL-10 in Patients with AdenomyosisObjective:To demonstrate the expression of IL-10,an immunotolerance related molecule,in the eutopic and ectopic endometrium of patients with adenomyosis;to compare the different levels of IL-10 in endometrium between normal women and patients with adenomyosis.To illustrate the role of IL-10 in establishing the state of immunotolerance and the possible contribution of IL-10 to understanding the pathogenesis of adenomyosis.Methods:The surgical biopses from 34 patients with adenomyosis were collected as experimental group,30 biopses normal endometrium collected as control groups,Immunohistological staining and hybridization in situ were used to detect the protein and mRNA of HLA-G in normal endometrium,eutopic and ectopic endometrium of adenomyosis.Quantitative analysis of IL-10 expression was performed according to H score system.The software of SPSS13.0 was used to analyze the experimental data.Results:1.In 30 cases normal endometrium,epithelial and stromal IL-10 expression was observed throughout the menstrual cycle.The epithelial cells of the endometrium showed higher staining intensity than the stromal cells in both proliferative and secretory phase samples.The epithelial and stromal staining was mostly cytoplasmic.The H score values of epithelial IL-10 immunostaining were significantly higher in the secretory phase than in the proliferative phase(P<0.01), whereas no difference was observed in stromal 1L-10 immunoreactivity throughout the menstrual cycle(P=0.62).2.Epithelial and stromal IL-10 immunoreactivity was observed in 34 eutopic endometrium and adenomyosis foci of women with adenomyosis in both proliferative and secretory phases.Similar to the normal endometrial samples,the epithelial cells of the eutopic endometrium and adenomyosis foci showed higher staining intensity than the stromal cells.The epithelial and stromal staining was mostly cytoplasmic.In the eutopic and ectopic endometrium of women with adenomyosis,epithelial cells showed higher staining intensity than those of the normal controls(both P<0.01).However,no significant difference was observed in the epithelial IL-10 mmunostainingH score values between the eutopic endometrium and adenomyosis foci(P=0.08).In addition,no differences were observed in stromal IL-10 expression among adenomyosis foci,their eutopic endometrium,and normal controls(all P>0.05).We observed a cyclic variation in eutopic epithelial IL-10 immunoreactivity throughout the menstrual cycle,with higher H score values in the secretory phase than in the proliferative phase(P<0.01).However,no significant difference was observed in eutopic stromal IL-10 immunoreactivity throughout the menstrual cycle in eutopic endometrium(P=0.22).3.A good correlationship was found between epithelial IL-10 expression in ectopic and eutopic endometrium in patients with adenomyosis(r=0.55,P<0.0001),however,no similar correlationship was found in stromal cells(r=0.02,P>0.05).4.The positive rate for IL-10 mRNA in normal endometrium,eutopic endometrium and ectopic endometrium is 28/30,33/34,34/34,respectively.The positive signal mostly localized in epithelial cells and stromal cells have weak signal.Conclusions:1.The higher level of IL-10 protein is found in the eutopic and ectopic endometrium of patients with adenomyosis,whereas the lower level was found in normal endometrium.These findings indicate that IL-10 may involve in the pathogenesis of adenomyosis by establishing the local microenviroment of immunotolerance and immunosuppression.2.The good correlationship between IL-10 expression in eutopic and ectopic endometrium in patients with adenomyosis indicates that the characterism of eutopic endometrium may play an important role in the pathogenesis of adenomyosis. PARTâ…¢Expression and Correlation of HLA-G and IL-10 in Ovarian Endometriotic Cyst FluidObjective:To demonstrate the expression of HLA-G and IL-10,the immunotolerance related molecule,in ovarian endometriotic cyst fluid.To illustrate the role of HLA-G and IL-10 in establishing the state of immunotolerance and the possible contribution of HLA-G and IL-10 to understanding the pathogenesis of endometriosis.To illustrate the correlation between the two molecules.Methods:The ovarian endometriotic cyst fluid and peritoneal fluid from 15 patients with endometriosis were collected as experimental group,15 cases of peritoneal fluid from patients with myoma as control group and Western blotting was used to analyze the level of HLA-G and IL-10 in the cyst fluid and peritoneal fluid. The software of SPSS13.0 was used to analyze the experimental data and t test was used to analyze the correlationship between the two molecules.Results:1.The peritoneal HLA-G was found neither in 15 patients with myoma nor in 13 patients with endometriosis and 2 cases HLA-G positive;whereas this molecule was found in high level in the ovarian endometriotic cyst fluid.2.The peritoneal IL-10 was found in patients with endometriosis but not in patients with myoma;whereas this molecule was found in high level in the ovarian endometriotic cyst fluid.3.A good correlationship was found between HLA-G and IL-10 expression in ovarian endometriotic cyst fluid(r=0.78,P<0.0001).4.A good correlationship was found between HLA-G and IL-10 expression not only in eutopic epithelial cells of adenomyosis(r=0.66,P<0.001),but also in ectopic epithelial cells of adenomyosis (r=0.60,P<0.001).Conclusions:1.The high levels of HLA-G and IL-10 in ovarian endometriotic cyst fluid indicate that the two molecules may involve in the pathogenesis of endometriosis by establishing the local microenviroment of immunotolerance and immunosuppression.2.The expression of HLA-G correlated with the expression of IL-10 in ovarian endometriotic cyst fluid.3.The expression of HLA-G correlated with the expression of IL-10 in adenomyosis.