Project1:Preliminary Study Of TCR-ζ Chain In The Pathogenesis Of Primary Sj(o|¨)gren’s Syndrome Project2:Significance Of Anti-α-fordrin Polypeptide Antibody In Patients With Sj(o|¨)gren’s Syndrome

Background and objectivePrimary Sjogren’s syndrome (SS) is a chronic inflammatory and lymphoproliferative autoimmune disease, characterised by progressive mononuclear cell infiltration within exocrine glands. Dryness of the mouth (xerostomia) and eyes (keratoconjunctivitis sicca) are the main organ-related clinical features. The pathogenesis of pSS is mainly due to abnormal activation of T/B lymphocytes in the body, which could leads to disorder of the balance between pro-inflammatory and anti-inflammatory. It is a question that whether weaken or even eliminate this imbalance is the key to treatment. The latest research reveals that the changes of expression and function of TCR-ξchain are associated with chronic inflammation in autoimmune diseases. The pathogenesis of pSS has not been clarified. Can we consider TCR-ξ chain as an ideal target for correcting imbalance? This study was designed to investigate the role of TCR-ξ chain in pSS by using cell line and clinical specimens. We want to investigate whether TCR-ξ chain is a potential target for the treatment of pSS, and provide certain theoretical basis and evidence for further study.Methods1. Preliminary research on the regulating mechanism of TCR-ξ chain in the T cell lineWe used Jurkat T cell line as the model of cell stimulation experiment. CD3monoclonal antibody was used to stimulate the Jurkat cell, and we monitored the changes of TCR-ξ chain’s level with the progress of the time. Western blot was used to determine the changes of TCR-ξ chain.2. Preliminary discussion of TCR-ξ chain in the patients with pSS Based on the collection of peripheral blood of pSS patients and healthy controls, flow cytometry was used to analysis the level of TCR-ξ chain in T lymphocytes. Statistical analysis was then carried on.3. Preliminary discussion of IL-33in the patients with pSS and conjoint analysis the clinical relevant indicators We collected serum of pSS patients and healthy controls, and tested the level of IL-33by ELISA. We use statistics to analysis the relationship among TCR-C chain, IL-33and other clinical indexes, such as blood sedimentation, immunoglobulin, complement, etc. The purpose of this part is to find the relationship among all these indicators to explore that whether TCR-ξ chain has a representative meaning as a potential point.Results1. As the growth of the time, under the stimulus by CD3monoclonal antibody, the level of TCR-ξ, chain in Jurkat cells was gradually decreased. It prompted the trend of adjustment of TCR-ξ chain in the condition with cell abnormal activation.2. Our research includes42patients in pSS group, the average age was45.0±10.6y;41cases in control group, the average age was40.2±6.6y. There were no statistical differences between age and gender of these two groups. The average level of CD3+CD247+cell in peripheral blood of pSS group was40.7±30.1(%), the other’s was57.1±17.8(%). Statistical analysis showed that the levels of TCR-ξ chain existed significant difference (P<0.01) between pSS patients and normal person.3. Our Study detected42pSS patients, IL-33level was103.4±72.3(pg/ml);16cases of healthy control group, the level of IL-33was72.4±40.6(pg/ml). We compared two of those three groups respectively:the differences of IL-33’s level were statistically significant between pSS group and control group(P<0.01). According to the value of ESR, the level of Immune globulin and complement in blood serum, we divided the patients into different tranches. And we use numerical indexs to analyse the correlation between them. We found that the level of IL-33may have relationship with ESR/IgG/C3, when it had no correlation with the level of TCR-ξ chain.Conclusion1. TCR and CD3formed the TCR-CD3complex through the salt bridge. TCR-ξ chain related to this complex is located in the cytoplasm of T cells, and its functional domains involved in the signal transduction in cells, which can induce the activation of T cells and lead to the happening of the disease. By using cell line in vitro stimulation experiments and western blot experiments, we could say that CD3monoclonal antibody stimulation, the activation of T cells and the levels of TCR-ξ chain were directly linked.2. The onset of pSS process is an disorders of anti-inflammatory and proinflammatory balance, and finally the abnormal activation of T/B cells happens as a result. TCR-ξ chain plays an important role in connecting during the abnormal activation process of T cells. Flow cytometry was uesd to detecting the level of TCR-ξ chain in fresh peripheral blood in all pSS patients and healthy volunteers. There were big differences between the two groups, suggesting that TCR-ξ chain play an important role in the process of T cells abnormal activation in the process of pSS disease.3. Interleukin1family plays an important role in many inflammatory diseases or infectious diseases in host defense and immune regulation. IL-33is a new factor of secretory cells of the family, which has the effect as pro-inflammatory factor and participates in the Th2cell mediated immune response. Our studies suggest that IL 33have the participation of the process of inflammation in pSS. We complete the clinical data, and analysis the results and some indicators commonly used for clinical evaluation. We want to find whether TCR-ξ chain could correctly reflect the extent of disease, and its relevance to the clinical indicators used commonly. But we only have found obvious correlation among IL-33, ESR, IgG and C3. We have not found any correlation between TCR-ξ chain and other indicators. BackgoundPrimary Sjogren’s Syndrome (pSS) is a systemic rheumatic disease characterized progressive lymphocytic and plasma cell infiltration of the salivary and lachrymal glands, and the presence of several auto antibodies in the blood. What’s more, it may be associated with chronic inflammatory systemic damage and result in multiple organ injury. The incidence of pSS and other common rheumatic disease, like rheumatoid arthritis, may be roughly equal, even higher. It has been lacking an ideal serological marker with high specificity and high sensitivity in clinic, at the same time the labial gland biopsy is too invasive to implement for patients in China. The poor inspection compliance, the internality of onset, and the lack of cognition of pSS all made clinical diagnosis of pSS a difficult problem. The characterization and identification of pathogenetically disease-relevant auto antigens is a key issue in autoimmunity. Serodiagnostic techniques of auto antibodies can be used in diagnosing pSS. Though the ANA, anti-SSA antibody and anti-SSB antibody are important in diagnosing pSS, the sensitivity and specificity is limited. In recent years, several studies suggested that anti-a-fordrin polypeptide antibody is of value in the diagnosis of SS and serves as a marker of disease activity. To evaluate the potential of anti-a-fordrin polypeptide antibody, the patients in this study all had received anti-a-fordrin polypeptide antibody detection in Lab of Rheumatology. ObjectivesTo investigate the significance of anti-a-fordrin polypeptide antibody detection in diagnosis of connective tissue disease and based on which to arouse further recognition and attention to the anti-α-fordrin polypeptide antibody detection in clinic.MethodsA cohort of105patients who had been under regular medical supervision at the Department of Rheumatology, Wuhan Tongji hospital, was evaluated in this study. All of them had received anti-a-fordrin polypeptide antibody detection in Lab of Rheumatology, Tongji hospital and were divided into four groups by discharge diagnosis. Statistical analysis was carried out in different groups, including clinical and laboratory features.Results1. The specificity of anti-a-fordrin polypeptide antibody screening in SS patients was67.1%, the sensitivity was40%. The specificity was higher than that of anti-SSA antibody (35.7%) and anti-SSB antibody (61.4%), while the sensitivity was lower than either anti-SSA antibody (85.7%) or anti-SSB antibody (71.4%).2. Among the four groups, the results of anti-SSA antibody (P<0.01), anti-SSB antibody(P<0.01), anti-ENA polypeptide antibody repertoire (P<0.01), IgA (P<0.05) and salivary gland ECT(P<0.01) were significantly different.3. The differences were also statistically significant when combining anti-a-fordrin polypeptide antibody and anti-SSA antibody, anti-SSB antibody and (or) salivary gland ECT for analysis (1P<0.01、2P<0.05). Conclusions1. For the diagnosis of primary SS, anti-a-fordrin polypeptide antibody detection had lower positive rate than SSA antibody and SSB antibody.2. ECT scan of salivary gland provided important reference to the diagnosis of SS, and is easier to carry out than traumatic labial gland biopsy.3. Histopathology examination of labial gland is still the golden standard of SS diagnosis.4. Detection of multiple auto antibodies can help to confirm the diagnosis of SS.