Expression Of T-bet Mrna,gata-3 MRNA On Peripheral Blood Mononuclear Cells Of HSP Patients

Objective:Henoch-Schonlein purpura (HSP) is a kind of small blood vessel vasculitis which multiple organs can be involved in. It occurs mainly in children. It belongs to the small systemic allergic vasculitis. Its pathologic basis is extensively small blood vessel vasculitis. The Major clinical characteristics of this disease are cutaneous purpura, abdominal pain, hematochezia, arthralgias and proteinuria. The etiology and pathogenesis of HSP are not clear now. Infection, foods, drugs et al can be sensitive factors,these make the sensitive individuals appear allergies and cause a series of injuries.HSP is a kind of systemic immunological vasculitis. In the recent years, the basic and clinical studies found that the serum of HSP patient possesed increased IgA and the IgA immune complex (IgA - IC) deposited on the blood vessel wall. It suggested that the IgA mediated immunological reactions were relevant to the pathogenesis of this disease. Now, the high immune globulin of HSP is usually taken to be as the result of B lymphocytes polyclonal activation. Then, there were lots of IgA immune complex to deposite on small systemic blood vessel wall and cause vasculitis. Multiple organs can be involved in, and there would be all sorts of clinical manifestations. The differentiation of T helper cells (Th cells) decides the nature of immunological responses. The Th1 cells promote the cellular immunity, and resist infections through reinforcing phagocytosis; Th2 cells mediate the humoral immunity by promoting B lymphocyte differentiation, proliferation and secreting antibody. Therefore, scholars thought that its pathogenesis was correlated with the disorder of function of T lymphocyte subpopulations.Th1 cells and Th2 cells play a key role in immunological responses. Cellular differentiation requires the precise action of the lineage-determining transcription factors. In the immune system, the CD4+ T helper cells can differentiate into distinct subpopulations, Th1, Th2, T-regulatory cells and Th17. The T- bet and GATA-3 are the key and specific transcription factors in the cell who decide the differentiation of the Th0 cell to Thl cell or Th2 cell. The T-bet and other members of the T-box family are defined by the highly conserved T-box DNA-binding domain. The domain is large, consisting of 189 amino acids. In the immune system, the T-bet is required for Th1 cell differentiation and the studies of the role of T-bet in Th1 development have been informative. The transcription factor GATA-3 is a main member of the GATA family. The GATA-3 is best known to the function as a master regulator of differentiation of Th2. If the GATA-3 is conditionally deleted in the immune system, the Th2 cells fail to develop. It is clear that the GATA-3 is important in the Th2 cells development. The study aimed to investigate the expression of the T-bet mRNA,GATA-3 mRNA in patients'peripheral blood mononuclear cells (PBMC) with HSP and to explore their possible significance by Real-time fluorescence quantitative polymerase chain reaction (RT-PCR).The real time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) is a kind of new technology which is based on traditional PCR detection method.In the RT-PCR processes, the amplification target sequence and fluorescence signals are detected simultaneously. The RT-PCR instrument collects all the fluorescent signals. After that, it automatically analysis the Ct value of every sample according to the mathematical algorithm through software. The Ct value is deducted the background signals and fluorescence threshold is set. It effectively solves the limitation that the traditional PCR only detects quantity at end-point. The RT-PCR posses the characteristics of high specificity, reliability, the stability of experiment results, repeatability etc.Methods:32 patients of HSP were involved in who were from the department of dermatology in the second hospital of Hebei Medical University. Among them, there were 14 cases in the simple group (only rash) and 18 cases in the mixed group (joint, the gastrointestinal tract or kidney involvement in one or more).The males and female were 14 and 18 cases.The average age was 26.9士0.5 years. The course of the disease was from 2 days to 1 year. 13 cases had a clear history of infection before the onset; 19 cases were without a history of infection. Inclusion criteria of patient group: (1) all the patients were with the feature of typical skin purpura, with or not abdominal pain, joint pain, hematuria; the platelet count, function and blood coagulation were normal. (2) Not accompanied with autoimmune diseases, allergic diseases, cancer or other serious systemic diseases(.3)Not used glucocorticoid or other drugs that affect the immune function of the body within 1 month before the test. 15 healthy persons were involved in the control group without allergic diseases, autoimmune diseases and family history of them. The gender and age were not statistically different between the control group and the patient group.Peripheral venous blood was obtained from the patients and the control group. the expressions of T-bet mRNA, GATA-3 mRNA in peripheral blood mononuclear cells (PBMC) were detected by the RT-PCR. Using the SPSS13.0 statistical software, all the experiment data was tested by the normality test to clear the general distribution types. The normal distribution data was with mean + standard deviation said (x + s) and the comparison between groups used the t test; Non-normal distribution data was with median (quarterback spacing) [Md ( Interquartile Range) ] and the comparison between groups used Wilcoxon test. When P < 0.05, we think that it is as a statistical significance.Results:1 Compaired with the normal controls, the expression of the T-bet mRNA decreased in HSP patients with a statistical significance (the mean ranks ofΔCt in HSP group and the normal controls respectively were 27.70 and 16.10, unilateral test P<0.05); while the expression of the GATA-3 mRNA increased with a statistical significance (the mean ranks ofΔCt in HSP group and normal controls respectively were 18.25 and 36.27, unilateral test P<0.05).2 In HSP patients, compared the simple group with the mixed group, the difference of the expression of the T-bet mRNA showed not statistically significant; (the mean rank ofΔCt, the simple group was 15.86 and the mixed group was 17.00, unilateral test P > 0.05), the expression of the GATA-3 mRNA in the mixed group of HSP increased with a statistical significance. (the mean in simple group was 3.11士0.63 , in mixed group was 2.20士0.78, P < 0.05) .Conclusisons : On the transcriptional level, this study hints that the immune disorder of the HSP posses highest potential of predominance in the Th2 lymphocyte Subpopulations, performing humoral immune hyperfunction. In HSP patients, compared the simple group with the mixed group, the expression of GATA-3 mRNA is increased in the mixed group. In return, it makes that the T-bet mRNA expression is decreased relatively in the mixed group. Then, the unbalance of Th1 / Th2 lymphocyte Subpopulations is more severe. It may have a close relation with the severity of the disease.