Th1/Th2/Th17/Treg Cytokines Expression Levels In The Peripheral Blood Of Type1and Type2Diabetes Patients

BackgroundWith the rapid economic development and the improvement of living standards, increament of environmental pollution, the incidence of diabetes is increasing rapidly. A latest epidemiological survey indicated that the diabetes prevalence in Chinese adults aged18and older was11.6%, and the prevalence of male is12.1%, the prevalence of female is11.0%, the prevalence of urban residents is14.3%, the prevalence of rural residents is10.3%. Diabetes is a common endocrine diseases which characterized by high blood sugar, caused by the interaction of genetic and environmental factors, inadequate or absolute secretion of insulin as well as lower target tissue sensitivity to insulin. The common symptoms of diabetes are polydipsia, polyuria, polyphagia and emaciation, at the same time it cause a series of metabolic disorder syndrome, such as the disturbance among protein, fat, water and electrolytes, it even concurrent eyes, kidney, nerve, cardiovascular and other viscera chronic damage.Type1diabetes can be divided into type1A and type1B.1A diabetes is mainly due to autoimmune β-cell destruction, leading to absolute insulin deficiency; while the mechanism of type1B diabetes is unknown and it has a tendency to ketoacidosis. Type1diabetes is a T cell-mediated organ-specific autoimmune disease, mainly due to the disorder of immune system, the destruction of islet β cells and absolute lack of insulin. T1DM onset at any age, and is mainly seen in children and adolescents, the incidence increased year by year. By2020, the prevalence of children under five years will increase1times. Numerous studies show that (3cell autoantigens, macrophages, dendritic cells, B lymphocytes, T lymphocytes and other related cytokines, chemokines are involved in its pathogenesis. A large number studies shown that people with type1diabetes is often non-single, it also associated with other autoimmune disease, such as autoimmune thyroid disease (AITD). The research shows that15-30%of type1diabetes patients also suffer from autoimmune thyroid disease,4-9%of type1diabetes patients with celiac disease,0.5%had Addison’s.CD4+T lymphocytes is the primary member of body’s immune system, may play an important role in the development of type1diabetes and type2diabetes. In the past, CD4+T cells are classified into two subsets, Th1and Th2. Thl cells secrete IFN-y, TFN-a, IL-2and other cytokines, mediate cell immune response; Th2cells mainly express IL4, IL-10and1L-13cytokines, increase B cell mediated humoral immune. In recent years, studies reported that a new CD4+T lymphocytes subgroup was founded, which can produce interleukin-17, named Th17subgroup. Th17is differ from Thl and Th2in differentiation, signal transduction and biological function. Th17can secrete IL-17, IL-21, IL-22, acting on different target cells, inducing the production of other cytokines, triggering inflammatory transmitter release. It plays an importment role in the development of proinflammatory response, mediated chronic inflammation, defense extracellular bacterial infections, mediated autoimmune diseases and so on. There is a close relationship between the Th17subgroup and autoimmune diseases, studies show that Th17subgroup participate in systemic lupus erythematosus, inflammatory bowel disease, experimental autoimmune encephalomyelitis and other autoimmune diseases. Type1diabetes as a common autoimmune disease, CD4+T lymphocyte subgroups’ imbalance invole in its onset. In recent years immunological prevention of diabetes has become a hot topic.ObjectiveTo detect the plasma concentrations of some cytokines expressed by relevant Th1, Th2, Th17, Treg in patients diagnosed with type1diabetes mellitus (T1DM) and type2diabetes mellitus(T2DM), and to detect the positive rate of glutamic acid decarboxylase antibodies(GAD-Ab), insulin autoantibodies(IAA) and islet cell antibodies(ICA) who were diagnosed with type1diabetes(T1DM) and autoimmune thyroid disease(AITD). Analysis the clinical characteristics of these patients, and to explore the use of these cytokines in the development of TIDM and T2DM.MethodsA total of180patients were included in the study. All these patients were divided into three groups.63patients diagnosed with type1diabetes mellitus,60patients diagnosed with type2diabetes mellitus, control group of57cases. Collected the peripheral blood, sandwich enzyme-linked immunosorbent assay for detection of IFN-y, IL-4, IL-10, IL-17. Detect islet-related antibodies, thyroid-associated antibodies. In the late, also detect the cytokines using flow cytometry.Results1、The results of enzyme linked immunosorbent assay for the detection of IFN-y, IL-4, IL-10, IL-17, as follows:In type1diabetes group, IFN-y and IL-17expression levels are higher than the healthy control group [(400.95±51.44) vs (340.99±39.47) pg/ml,(26.87±2.47) vs (19.41±2.80) pg/ml, respectively, t=6.49、14.29, P<0.01]; IL-4and IL-10are lower than the healthy control group [(21.63±4.22) vs (31.49±2.42) pg/ml,(15.69±3.22) vs (24.13±2.17) pg/ml, respectively, t=-14.76、-15.75, p<0.01].In type2diabetes mellitus control group, IFN-γ and IL-10expression levels are lower than the healthy control group [(303.67±50.33) vs (340.99±39.47) pg/ml,(22.49±2.95) vs (24.13±217) pg/ml, respectively, t=-3.77,-2.9, P<0.01]; IL-17is higher than the healthy control group [(21.50±2.15) vs (19.41±2.80) pg/ml, t=3.71, P<0.01]; IL-4is higher than the healthy control group [(32.69±4.10) vs (31.49±2.42) pg/ml, t=1.55, P>0.05].Meanwhile, IFN-y and IL-17of T1DM group are higher than these of T2DM group (respectively, t=8.93、10.69, P<0.01); IL-4and IL-10of T1DM group are lower than these of T2DM group (respectively, t=-12.38,-10.22, P《0.01)To compare with the group of T1DM whose disease course≤2years, the group of T1DM whose disease course>2years has a higher level of IFN-y, and lower levels of IL-17、IL-4、IL-10, but the differences are not statistically significant (p>0.05). The T2DM control group whose course≤2years, the levels of IFN-y and IL-17are higher than the T2DM control group whose course>2years,and the levels of IL-4and IL-10are lower,but the differences are not statistically significant (p>0.05).Meanwhile, in diabetic patients, the serum levels of IL-17and IFN-y expression positively correlated (r=0.715, P<0.05), IL-17and IL-10are negatively correlated (r=-0.61, P<0.05).In the group any one islet-related antibodies positive, TPO-Ab, TG-Ab, TR-Ab positive rates were higher, the difference was statistically significant (P<0.05), but the level of cytokines were not statistically significant. T1DM group serum IL-17 levels and HbAlc were positively correlated.(r=0.59, P<0.05).2、The results of using flow cytometry for the detection of IFN-y, IL-4, IL-17, as follows:Type1diabetes group Th17/CD4+T cells (%)[2.30(1.65/4.30)] is higher than type2diabetes [2.00(1.08/2.88)], higher than the control group [1.15(0.63/1.70)], Kruskal-Wallis H test results are shown H=8.54, P<0.05.Type1diabetes group Thl/CD4+T cells (%)[15.55(13.25/17.47)] is higher than the type2diabetes group [14.55(11.78/15.38)], higher than the control group [10.75(8.50/13.45)], Kruskal-Wallis H test results are shown H=9.31, P<0.05.Type1diabetes group Th2/CD4+T cells (%)[3.35(2.37/5.65)] lower than the control group [5.30(3.85/7.40)], lower than type2diabetes [5.40(2.92/7.70)], Kruskal-Wallis H test results are shown H=4.33, P>0.05.ConclusionThe subsets of CD4+T cell in T1DM and T2DM are imbalance, Thl/Th2and Th17/Treg are imbalance.Type1diabetes mellitus based on cellular immunity and autoimmune inflammatory, accompanied humoral immune defects. Type2diabetes may exist cellular immune deficiency, autoimmune inflammatory also involved in its development. Different course of diabetic patients exhibit different cytokines dominant trend. Meanwhile, in diabetic patients, the serum levels of IL-17and IFN-y expression positively correlated, IL-17and IL-10are negatively correlated. T1DM group serum IL-17levels and HbAlc were positively correlated. In type1diabetes patients any one islet-related antibodies positive, have higher rates of thyroid-associated antibodies, may have a higher risk of autoimmune thyroid disease.