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

Background and objectivePrimary Sj gren’s syndrome (SS) is a chronic inflammatory andlymphoproliferative autoimmune disease, characterised by progressive mononuclearcell infiltration within exocrine glands. Dryness of the mouth (xerostomia) and eyes(keratoconjunctivitis sicca) are the main organ-related clinical features. Thepathogenesis of pSS is mainly due to abnormal activation of T/B lymphocytes in thebody, which could leads to disorder of the balance between pro-inflammatory andanti-inflammatory. It is a question that whether weaken or even eliminate thisimbalance is the key to treatment. The latest research reveals that the changes ofexpression and function of TCR-ζchain are associated with chronic inflammation inautoimmune diseases. The pathogenesis of pSS has not been clarified. Can we considerTCR-ζ chain as an ideal target for correcting imbalance? This study was designed toinvestigate the role of TCR-ζ chain in pSS by using cell line and clinical specimens.We want to investigate whether TCR-ζ chain is a potential target for the treatment ofpSS, and provide certain theoretical basis and evidence for further study.Methods1. Preliminary research on the regulating mechanism of TCR-ζ chain in the Tcell 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-ζ chain’s level with the progress of the time. Western blot wasused to determine the changes of TCR-ζ chain.2. Preliminary discussion of TCR-ζ chain in the patients with pSSBased on the collection of peripheral blood of pSS patients and healthy controls,flow cytometry was used to analysis the level of TCR-ζ chain in T lymphocytes.Statistical analysis was then carried on.3. Preliminary discussion of IL-33in the patients with pSS and conjoint analysisthe clinical relevant indicatorsWe collected serum of pSS patients and healthy controls, and tested the level ofIL-33by ELISA. We use statistics to analysis the relationship among TCR-ζ 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 theseindicators to explore that whether TCR-ζ chain has a representative meaning as apotential point.Results1. As the growth of the time, under the stimulus by CD3monoclonal antibody, thelevel of TCR-ζ chain in Jurkat cells was gradually decreased. It prompted the trendof adjustment of TCR-ζ 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 statisticaldifferences between age and gender of these two groups. The average level ofCD3+CD247+cell in peripheral blood of pSS group was40.7±30.1(%), the other’swas57.1±17.8(%). Statistical analysis showed that the levels of TCR-ζ chainexisted 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 levelwere statistically significant between pSS group and control group(P<0.01).According to the value of ESR, the level of Immune globulin and complement inblood serum, we divided the patients into different tranches. And we usenumerical indexs to analyse the correlation between them. We found that the levelof IL-33may have relationship with ESR/IgG/C3, when it had no correlation withthe level of TCR-ζ chain.Conclusion1. TCR and CD3formed the TCR-CD3complex through the salt bridge. TCR-ζ chainrelated to this complex is located in the cytoplasm of T cells, and its functionaldomains involved in the signal transduction in cells, which can induce the activationof T cells and lead to the happening of the disease. By using cell line in vitrostimulation experiments and western blot experiments, we could say that CD3monoclonal antibody stimulation, the activation of T cells and the levels of TCR-ζchain were directly linked.2. The onset of pSS process is an disorders of anti-inflammatory and proinflammatorybalance, and finally the abnormal activation of T/B cells happens as a result. TCR-ζchain plays an important role in connecting during the abnormal activation processof T cells. Flow cytometry was uesd to detecting the level of TCR-ζ chain in freshperipheral blood in all pSS patients and healthy volunteers. There were bigdifferences between the two groups, suggesting that TCR-ζ chain play an importantrole in the process of T cells abnormal activation in the process of pSS disease.3. Interleukin1family plays an important role in many inflammatory diseases orinfectious diseases in host defense and immune regulation. IL-33is a new factor ofsecretory cells of the family, which has the effect as pro-inflammatory factor andparticipates in the Th2cell mediated immune response. Our studies suggest that IL– 33have the participation of the process of inflammation in pSS. We complete theclinical data, and analysis the results and some indicators commonly used forclinical evaluation. We want to find whether TCR-ζ chain could correctly reflect theextent of disease, and its relevance to the clinical indicators used commonly. But weonly have found obvious correlation among IL-33, ESR, IgG and C3. We have notfound any correlation between TCR-ζ chain and other indicators. BackgoundPrimary Sj(?)gren’s Syndrome (pSS) is a systemic rheumatic disease characterizedprogressive lymphocytic and plasma cell infiltration of the salivary and lachrymalglands, and the presence of several auto antibodies in the blood. What’s more, it maybe associated with chronic inflammatory systemic damage and result in multiple organinjury. The incidence of pSS and other common rheumatic disease, like rheumatoidarthritis, may be roughly equal, even higher. It has been lacking an ideal serologicalmarker with high specificity and high sensitivity in clinic, at the same time the labialgland biopsy is too invasive to implement for patients in China. The poor inspectioncompliance, the internality of onset, and the lack of cognition of pSS all made clinicaldiagnosis of pSS a difficult problem. The characterization and identification ofpathogenetically disease-relevant auto antigens is a key issue in autoimmunity.Serodiagnostic techniques of auto antibodies can be used in diagnosing pSS. Thoughthe 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 thatanti-a-fordrin polypeptide antibody is of value in the diagnosis of SS and serves as amarker of disease activity. To evaluate the potential of anti-a-fordrin polypeptideantibody, the patients in this study all had received anti-α-fordrin polypeptide antibodydetection in Lab of Rheumatology. ObjectivesTo investigate the significance of anti-α-fordrin polypeptide antibody detection indiagnosis of connective tissue disease and based on which to arouse further recognitionand attention to the anti-α-fordrin polypeptide antibody detection in clinic.MethodsA cohort of105patients who had been under regular medical supervision at theDepartment of Rheumatology, Wuhan Tongji hospital, was evaluated in this study. Allof them had received anti-α-fordrin polypeptide antibody detection in Lab ofRheumatology, Tongji hospital and were divided into four groups by dischargediagnosis. Statistical analysis was carried out in different groups, including clinical andlaboratory 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-SSAantibody (35.7%) and anti-SSB antibody (61.4%), while the sensitivity was lowerthan 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-SSBantibody(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-fordrinpolypeptide antibody and anti-SSA antibody, anti-SSB antibody and (or) salivarygland ECT for analysis (1P<0.01、2P<0.05). Conclusions1. For the diagnosis of primary SS, anti-a-fordrin polypeptide antibody detection hadlower 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 SSdiagnosis.4. Detection of multiple auto antibodies can help to confirm the diagnosis of SS.