The Joint Detection Of T Helper Cell Subsets In Systemic Lupus Erythematosus Disease Assessed Value

BackgroudSystemic lupus erythematosus (SLE) is a prototypic systemic disease characterized by autoimmune involved autoimmune inflammation and, SLE multisystem-involved multi-organ-damaged autoantibody production (antinuclear antibody, ANA). It is a chronic processing, has a high death ratio. The etiology and pathogenesis mechanisms of SLE have not been clearly elucidated. At present, there is no specific effective treatment for SLE.There are two forms disorders of the cellular immune and humoral immune system in the course of SLE. T lymphocyte abnormalities and B lymphocyte excessive activation cause the abnormal immune system response. The above situations are main pathogenesis of SLE.In recent years,the immune disorders research of SLE tends to be thorough, multiple studies SLE itself that since lymphocyte balance is broken, excessive actiated T lymphocytes led to many cloned B cell differentiation and activation, resulting in a massive autoantibody cause multi-system damage involvement.The current research found that T lymphocyte exception may be the important factors SLE morbidity.One research has shown that the apoptosis of Th cells in SLE increased significantly, and apoptosis level and disease activity related.Therefore, the imbalance of T helper cells number and function is an important aspect of the immune function abnormality.T helper cells is not a single group of cells, but contains a series with various functions lymphocyte subsets.Naive CD4+ T lymphocytes may differentiate into four kinds of cells:T helper cell type 1(Th1),T helper cell type 2(Th2),T helper cell type 17(Th17)and regulatory T cells(Treg). Their cytokines and immune function each are not identical. Therefore, there are complicated relationships of the different functions of lymphocyte subsets in the differentiation and development and functional execution and other aspects. Need to study Th cell subtypes playing a role pathogenetic mechanisms in SLE, that can conduce to further clear understand the role of Th cells in SLE.Early, many research only focus on therelationship of Th1/ Th2 imbalance state and SLE.Imbalance in Th1/Th2 plays an import role in pathogenesis of SLE."Th2 prominence"and"Th1 prominence"were proved in animal model and lupus patients. But with the deepening of the research of SLE ,it was found that a lot of SLE clinical manifestations and laboratory characteristics cannot use Th1 / Th2 imbalances model to explain. In recent years ,the understanding of T helper cells subsets was more comprehensive. In 1995 and 2005, Harrington and Sakaguchi successively found the two new members: regulatory T cells (Treg) and Th17. Gradually, the important role of Th17 / Treg balance was discovered in SLE. Both with belong to Th cells, but Th17 and Treg are completely opposite in differentiation and biology performance. Th17 stimulate an autoimmune response and Treg cells can reduce autoimmune disease reaction. They maintain the immune state of relative stability.Th1,Th2,Th17,Treg contribute to the pathogenesis in RA, the imbanlance of Th1/Th2,Th17/Treg lead cell function disorder activation of B cells,increase the degree of SLE. Th17/Treg cells may be involved in the occurrence of disease and development,in some degree it can be the index of exultation the disease .thus Th1/Th2,Th17/Treg as a whole may be a more scientific than one single index.In short, this research from Th cell apoptosis, in-depth analysis the imbalance of Th1 / Th2, Th17 / Treg in SLE and related cytokines spectrum change characteristics. And we further study on Th17 / Treg specific transcription factors. Through the relatively large sample of SLE cases material, combined determination Th lymphocyte subsets cell, protein, gene level change and the corresponding SLE disease activity index, we discusses the relationship of Th lymphocyte subsets and SLE disease activity ,clinical symptoms of disease assessment and SLE's diagnosis value.Objective(1)To study the levels of lymphocyte apoptosis and disease activity index in systemic lupus erythematosus patients.(2)To study the balance of Th1/Th2,Th17/Treg and disease activity index in systemic lupus erythematosus patients.(3)To study the level of cytokines secreted by Th1/Th2,Th17/Treg and disease activity index in systemic lupus erythematosus patients.(4)To study the level of RORγt and Foxp3 and disease activity index in systemic lupus erythematosus patients.MethodsEighty-nine SLE patients,diagnosis complied with ACR classification standard ,were recruitedin the second hospital shanxi medical university and the 264 hospital of PLA from June 2009 to September 2010. We collected SLE patients generally and clinical materials.According to systemic disease activity index (SLEDAI) integral evaluation, patients were divided into light SLE, moderate activity type, heavy three groups. Meanwhile, 27 patients normal were selected as control group. Th apoptosis rate and Th1 / Th2, Th17 / Treg cell level were detected with Flow cytometery. Concentrations of IL-2,IL-4,IL-6,IL-10,TNF-α,IFN-γ,IL-17A and TGF-βin plasma were measured by Cytometric Bead Array (CBA) and Enzyme-linked immunosorbent assay (ELISA). Expression Levels of RORγt and Foxp3 mRNA were measured by RT-PCR. The data was analyzed by SPSS 13.0.Results(1)SLE patients had higher apoptosis rate of CD4+T cells than health controls[(1.11±0.36)% vs(.0.36±0.15)%,P<0.001]. Apoptosis rate of Th cells in serious group was higher than that in the light group ,medium group and control group[(1.51±0.31)%,(0.81±0.19)%,(1.03±0.21)%vs.(0.36±0.15), P <0.001].(2)The percentage of Th1 cells in peripheral lymphocyte in SLE was lower than that in the control, and there was no statistical significance in the percentage of Th2 cells between SLE and control. The ratio of Th1/Th2 in serious group was higher than that in the light group and medium group.Th2/CD4+ correlated positively with SLEDAI scores ( r=0.4012, P =0.0001).Th1/Th2 correlated negatively with SLEDAI scores(r=-0.2683,P =0.0130). There was no statistical significance in the percentage of Th17 cells between SLE and control. But the ratio of Th17/ CD4+ in SLE was higher than that in the control(P =0.012).The percentage of Treg cells in peripheral lymphocyte in SLE was lower than that in the control (P =0.023). The percentage of Th17,Th17/CD4+ in serious group were higher than that in the light group and medium group. The percentage of Treg cells in the serious group was lower than that in the light group and medium group.There were positive correlation between Th17,Th17/CD4+,Th17/Treg and SLEDAI scores(r=0.3608, P =0.0006, r =0.4522, P <0.001, r =0.4240, P <0.001).There was negative correlation between Treg and SLEDAI scores(r =-0.352, P =0.0010)(3)IL-17,IL-10,IL-6,IL-4 detected by CBA were elevated compared to those in healthy control (P<0.05),and the levels of other cytokines showed no statistic differences between patients with SLE and healthy control. The levels of plasma IL-17,IL-6 andIL-4 were positively correlated with SLE disease activity index(SLEDAI)scores. The levels of plasma IL-2,IFN-γshowed negative correlation with SLEDAI. No significant correlation were observed between plasma TNF,IL-10 and SLEDAI.The levels of plasma IL-17,IL-10,IL-6,IL-4 in the serious group were higher than that in the light group and medium group. The levels of plasma IFN-γ,IL-2 in the serious group were lower than that in the light group and medium group(F=52.660,P <0.001).The levels of TNF-αshowed no statistic differences between patients in serious group and other group. The levels of TGF-βdetected by ELISA showed no statistic differences between patients in SLE and healthy control(F=2.150,P =0.157). No significant correlation was observed between plasma TGF-βand SLEDAI(r =-0.071,P =0.5150).(4)The SLE patients showed higher levels of ROR-γt,Foxp3 mRNA than the healthy controls(t=-2.656,P =0.012;t =-2.873,P =0.007).Expression of ROR-γt mRNA in the serious group was higher than that in the light group and medium group(F=3.335,P =0.047). The levels of Foxp3 mRNA showed no statistic differences between patients and healthy controls(F=2.119,P =0.129). The levels of ROR-γt mRNA was positively correlated with SLEDAI and Th17(r =0.4969, P =0.0038, r =0.3960, P =0.0992) .No significant correlation was observed between ROR-γt mRNA and other Th cells and cytokines. No significant correlation was observed between Foxp3 mRNA and Th and cytokines realted. ConclusionThere is Th apoptosis abnormalities in SLE patients and Th apoptosis rate Th2/CD4+,Th17,Th17/CD4+,Th17/Treg were positively correlated with SLEDAI. Th1/Th2 and Treg were negatively correlated with SLEDAI. Concentration of IL-17,IL-6,IL-4 were positively correlated with SLEDAI , and IL-2,IFN-γwere negatively correlated with SLEDAI. ROR-gamma t mRNA was significantly correlated with the SLEDAI score and Th17. Our research shows that imbalance state of Th1 / Th2 and Th17/Treg was found in SLE., and united detection of Th lymphocyte subsets was possibly more meaningful for SLE condition evaluation.