| Recurrent spontaneous abortion(RSA) is a common disease in the department of obstetrics and gynecology, the incidence of which is about1-5%in the women of child-bearing age, and it seriously influence patient’s physical and mental health.The cause of RSA is complex, about50%of which is unknown, bringing certain difficulty to clinical prevention and control.In recent years, lymphocyte immunotherapy(LIT) has been widely used in unexplained recurrent spontaneous abortion(URSA) at home and abroad, but its curative effect is controversial continuously.In recent years, a large number of studies on the mechanism of LIT are discussed at home and abroad, making some achievements, but the specific mechanism is still not clear.In this text, we will analyze the etiological factors of RSA by collecting the clinical data of RSA patients.And then we will analyze the immune phenotype of RSA patients before and after LIT to investigate the mechanisms of LIT and discusses its application value for the diagnosis of RSA.This research will be divided into three parts:Part I The analysis of etiological factor of RSA patientsObjectiveTo analyze the etiological factors of RSA by collecting the clinical data of RSA patients and provide a reference basis for clinical diagnosis and therapy.Methods195cases of RSA patients were selected and scanned strictly for cause screening mainly including couple peripheral blood karyotype, infection factors, genital tract anatomy, endocrine and immune factors (including some autoantibody). Results195cases of RSA patients,181cases were primary RSA (92.82%),14cases were secondary RSA(7.18%);11cases with abnormal chromosome (5.64%);20cases with abnormal uterine anatomical structure (10.26%);11cases with genital tract infection (5.64%);3cases with positive TORCH-IGM (1.54%);37cases with abnormal endocrine (19.8%);19cases with positive autoantibody (10.2%);10of all the abnormal cases with two or more abnormal factors,104cases with unknown etiological factors (53.33%).ConclusionThe etiological factors of RSA are complicated, mainly including some differentiable factors:genetic factors, infection factors, uterine anatomical structure abnormal, endocrine and autoimmune factors. RSA is mainly related to immunological factors. However, the relationship between some factors and RSA has not been fully clear, bringing some difficulties to the diagnosis and treatment for RSA.Therefore, a detailed etiological screening for RSA patients is the premise and the key to successful treatment.Part Ⅱ The study of the immune phenotype of lymphocytes in patients with URSAObjectiveTo analyze the phenotypes of T、B、NK and Treg cells in peripheral blood of URSA patients in the non-pregnant state, to investigate the pathogenic immune basis of URSA and try to seek some specific immune indexes for the auxiliary diagnosis of URSAMethods25cases of URSA were selected as research group,25cases of normal early pregnancy women and70cases of normal not pregnancy women were selected as control groups. The phenotypes of T(CD3+、CD4+、CD8+、CD3+HLA-DR+、CD4+HLA-DR+、 CD8+HLA-DR+、CD4+/CD8+)、B(CD19+、CD5+CD19+、CD5-CD19+)、NK(CD3-CD56+、 CD56dimCD16+、CD56brightCD16-、CD56+CD69+)and Treg(CD4+CD25+CD127dim)cells in peripheral blood were determined by flow cytometry method.Results1. There was no significant difference in the proportion of CD3+T cells and the ratio of CD4+/CD8+in the three groups(P>0.05).The percentage of CD3+HLA-DR+T CD8+HLA-DR+T cells in URSA was significantly higher than that of NEP group(P<0.01).The percentage of CD4+T cells of URSA group and the CD8+T cells of NEP group was considerably higher than that of NNP group.2. The proportion of Treg cells in NEP group was remarkably increased than that of NNP and URSA groups(P<0.01), but there was no significant difference in NNP and URSA group.3. The percentage of CD5+CD19+B cells in lymphocyte of URSA group was increased than that of controls, but the proportion of CD5-CD19+B cells in B cells was decreased.4. The proportion of CD3-CD56+NK, CD56dimCD16+NK cells of URSA group was considerably higher than that of NEP group (P<0.05), but there was no obvious difference when compared with NNP group.The percentage of CD56brightCD16-NK in URSA group was significantly reduced than that of NEP and NNP groups (P>0.05), and the percentage of CD3+CD56+NKT and CD69+CD56+NK cells had no obvious difference in the three groups.Conclusions1. The immune imbalance still exist in URSA patients in non-pregnant state, the increase of CD19+B、CD56dim CD16+NK cells、the activation of CD8+T cells may be the pathogenic immune basis of URSA. Treg and CD8+T cells are conducive to maintaining pregnancy.2. Flow cytometry is a simple and practical technology, it could be applied to the auxiliary diagnosis of patients with URSA. The immune phenotype of lymphocyte, such as Treg cells、the activation of T cells and CD56dim、CD56bright NK cell could be the diagnostic indicators for URSA.Part III Analysis the change of lymphocyte immune phenotype before and after the active immunotherapy in patients with unexplained recurrent spontaneous abortionObjectiveTo investigate the phenotypes of T、B、NK and Treg cells in peripheral blood of URSA patients before and after LIT, and to study the possible mechanism of LIT, to provide theoretical basis for clinical effect assessment.Methods14URSA patients who would be treated with active immunotherapy were selected as study group.The phenotypes of T、B、NK and Treg cells in peripheral blood of URSA were detected by flow cytometry before and after LIT. The detection index was the same as part two.Results1. There was no obvious difference in the proportion of peripheral B cells and CD4+CD25+CD127dimTreg cells in URSA patients before and after LIT.2. The proportion of CD3CD56+NK cells in lymphocytes (11.06±4.14) was significantly reduced (9.10±4.07) after LIT (P<0.05).But there was no obvious difference in the proportion of CD56bright CD16-、CD56dimCD16+、CD3+CD56+and CD69+NK (activated) cells before and after LIT.3. The proportion of CD3+T cell in lymphocyte and HLA-DR+T cell in CD4+T cell was significantly increased, but the proportion of CD4+T cell in CD3+T cell is decreased after LIT(P<0.05). CD8+T cell in CD3+T cell、HLA-DR+T cell in CD8+T cell and the ratio of CD4+/CD8+had no significant difference after LIT.Conclusions1. LIT could decrease the proportion of CD3-CD56+NK cells and increase the activation and stimulation of CD4+T cell. The increase of CD3+T cells may means enhanced cellular immunity.But B cells and Treg cells did not change dramatically before and after LIT.2. The immune phenotypes of T cells and NK cells will probably be used as one of the important indicator of the therapeutic effect for URSA patients. |
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