Autoantibodies,T Lymphocyte Subsets And Cytokines In Patients With Premature Ovarian Insufficiency

Part IAutoantibodies in Patients with Premature Ovarian InsufficiencyBackground:Premature ovarian insufficiency(POI)is a gynecological endocrine disease with highly clinical heterogeneity.POI is one of the important causes for female infertility,and the etiology of POI is complicated.The known causes include genetic,immunological,infectious and iatrogenic factors,however,the etiology of most POI patients is still unclear.Previous studies have shown that autoimmune disturbance accounts for 4%-30%of POI,mainly including autoimmune oophoritis,associated autoimmune diseases and anti-ovarian autoimmune antibodies.Currently,there is no reliable immune test or diagnostic indicator for early diagnosis of autoimmune POI.The diagnosis of ovarian autoimmune abnormality depends on ovarian biopsy,however,it is difficult to be promoted in clinical practice because of its invasiveness.Serum autoantibodies may be used as markers of autoimmune POI.There have been a lot of studies on ovarian autoantibodies reported,but there are some controversies in terms of specificity,sensitivity and consistency of results.The ovaries and adrenals share common antigens,particularly those associated with steroid synthesis.Therefore,anti-adrenal antibody(AAA)may be a potential immune marker for POI risk prediction or diagnosis.However,the importance and role of AAA in idiopathic POI is unclear.Objective:The aim is to analyze the positive rates of anti-adrenal cortex antibody(AAA),anti-cardiolipin antibody(ACA),antinuclear antibody(ANA)and anti-double-stranded DNA(ds-DNA)in patients with idiopathic POI,and find potential immune markers of POI,explore the role of autoimmune in the pathogenesis of POI,and therefore provide theoretical basis for early prediction and diagnosis of POI.Methods:A total of 250 idiopathic POI patients and 256 healthy women were enrolled in this study.AAA was detected by indirect immunofluorescence,and ANA,ACA and ds-DNA were detected by enzyme-linked immunosorbent assay.The differences of clinical features between AAA-positive and AAA-negative POI patients were compared,and ovarian biopsy tissues were histologically observed in some POI patients.The adrenal function of 15 AAA positive POI patients was followed up to evaluate the risk of long-term adrenal insufficiency.Results:The positive rate of AAA in POI patients was 19.2%,significantly higher than that in control women(5.9%,P<0.01).The positive rate of ACA,ANA and ds-DNA was 0.8%,3.6%and 10.0%,respectively,and the difference was not statistically significant compared with the control group(0.4%,1.2%and 5.9%,P>0.05).It was suggested that there were autoimmune abnormalities in POI patients,and AAA could be used as a marker to predict and diagnose autoimmune POI.There was no significant difference in clinical characteristics,such as FSH,age at menarche,age at irregularity,and ovarian volume between AAA positive POI patients and AAA negative patients(P>0.05).Laparoscopic ovarian biopsy was performed in 13 POI(6 AAA positive and 7 AAA negative)patients,all of whom were found to have atrophic ovaries without ovarian follicles.15 AAA positive POI patients were followed-up for adrenal function,and only one patient developed adrenal insufficiency 3 years after the diagnosis of POI.Conclusion:In summary,the positive rate of AAA was significantly increased in POI patients,indicating that autoimmune abnormality was involved in the pathogenesis of POI,and the common immune mechanism between ovary and adrenal was also confirmed.AAA can be used as a marker for the prediction and diagnosis of ovarian autoimmune abnormalities,but the specific mechanism of AAA in the progression of immune POI disease still needs further research.Part ?Cytokines in Patients with Premature Ovarian Insufficiency Background:The imbalance of immune systems and decrease of immune surveillance may be the main pathogenic mechanism of autoimmune disorders.Abnormal cellular immunity plays an important role in the pathogenesis of POI,especially for dysregulated immune cells and related cytokines.It has been reported that cytokines,such as IFN-y,TNF-a and TGF-P involved in normal follicle development,ovulation and follicular atresia process by endocrine,paracrine and(or)autocrine mechanism,Meanwhile these cytokines can also induce B cell proliferation,differentiation and secretion of antibody,promote the differentiation of NK cells and CD8+T cell,and thus lead to ovarian autoimmune disorders.At present,Currently there are few researches on cytokines in POI patients with contradicted results and small sample size,not enough to conclude the true association with POI.Therefore,further research with larger sample size are needed to confirm the role of cytokines in POI.Objective:The aim is to detect the serum levels of some cytokines in POI patients,such as IFN-?,IL-1?,IL-6,IL-9,IL-17A,IL-22 and TGF-p1,analyze the differences of different cytokines between POI patients and normal healthy women,and investigate the role of cytokines in the pathogenesis of POI.Methods:A total of 100 women with idiopathic POI and 100 healthy age-matched women were recruited.The serum levels of cytokines,such as IFN-?,IL-1?,IL-6,IL-9,IL-17A,IL-22 and TGF-?1,were measured by enzyme-linked immunosorbent assay,and the differences were compared between women with POI and healthy women.Results:The serum levels of cytokine IL-22,IFN-y and IL-1? in POI patients were 107.25±16.01pg/ml,2.84(1.69-3.91)pg/ml and 4.46(1.94-23.42)pg/ml,respectively,both of which were significantly higher than those in the control group(P<0.05).The levels of TGF-?,IL-6 and IL-9 in POI patients were 11.43±6.23ng/ml,1.27(0.25-5.35)pg/ml and 0.33(0.20-0.40)pg/ml respectively,which were significantly lower than the control group(P<0.05).The serum level of IL-17A in POI patients was 0.77(0.57-1.02)pg/ml,which was not significantly different from that in healthy women(P>0.05).Conclusion:In summary,the abnormal expression of cytokines in patients with idiopathic POI indicates that cytokines play an important role in the pathogenesis of POI.The decrease of immunoregulatory cytokine TGF-? and increase of pro-inflammatory cytokines IFN-? and IL-22,might lead to continuous immune attack on local ovarian follicles and induce depletion of follicles and then POI occurs.Part?T Lymphocyte Subsets in Patients with Premature Ovarian InsufficiencyBackground:The etiology of premature ovarian insufficiency(POI)is heterogenous,and abnormal cellular immunity,especially the imbalance of CD4+ helper T cell and synergistic action of pro-inflammatory cytokines may be the important immunological mechanism of POI.T lymphocytes are involved in the development of autoimmune diseases by mediating cellular immunity.Previous studies showed that change of peripheral blood T lymphocytes in POI patients,especially the increase of CD4+T cells and decrease of effector CD8+/CD57+T cells(cytotoxic T lymphocytes),which suggests that Thl/Th2 imbalance was involved in pathogenesis of POI.With the development of immunology,new lymphocyte subsets have been identified and isolated,but few studies have been conducted in patients with POII.Currently there are few and controversial researches on lymphocyte subsets in patients with POI,therefore,the pathogenesis of dysregulated lymphocyte subsets in POI patients cannot be accurately and comprehensively elucidated.Objective:The aim is to analyze the proportion of Thl cells,Th17 cells,Th22 cells and Treg cells in the peripheral blood of POI patients at different clinical stages and to explore the role of cellular immunity in POI.Methods:A total of 38 idiopathic POI patients and 21 healthy age-matched women were enrolled in this study.POI patients were divided into POI group(FSH>40IU/L)and POF group(25IU/L<FSH<40IU/L)according to serum FSH level.The proportion of different lymphocyte subtypes,Thl(CD3+CD8-IFN-?+)cells,Th17(CD3+CD8-IL-17+)cells,Th22(CD3+CD8 IL-22+)cells and Treg(CD4+CD25+FOXP3+)cells in peripheral blood were measured by flow cytometric analysis and compared between different groups.Results:The proportion of Thl cells in POF group and POI group was(9.92±2.36)%and(10.15±2.29)%,respectively,which were significantly higher than that in control group(5.29±1.44)%.However,there was no significant difference between POF and POI groups.The proportion of Th17 cells in the POF group and POI group was(0.60±0.40)%and(0.67±0.39)%respectively,while(0.52±0.36)%in control group,and there was no significant difference between the three groups(P>0.05).The proportion of Th22 cells in POF group and POI group was(0.33±0.12)%and(0.32±0.12)%respectively,while(0.31±0.14)%in control group,and there was no significant difference between the three groups(P>0.05).The proportion of Treg cells in POF group and POI group was(2.44±1.13)%and(2.15±0.76%)respectively,which were both significantly lower than that in the normal control group(P<0.05),There was no significant difference between POF group and POI group(P>0.05).Conclusion:The increased proportion of Thl cells and the decreased proportion of Treg cells may play an important role in the pathogenesis of POI by mediating cellular immunity disturbance.However,the specific role of T lymphocyte subsets and related cytokines in the pathogenesis of POI still needs to be further discussed.