The Research Of Dexamethasone-induced Systemic Lupus Erythematosus Patients,cd4+cd25- Lymphocyte Apoptosis In Vivo And In Vitro

Objective To study the systemic lupus erythematosus (SLE) peripheral blood T regulatory cells (Treg) markers, as well as different Treg cells in the pathogenesis of SLE; to explore the relevance of CD127 and FoxP3;identificate CD4 + CD25 + CD127low/-T cell immunosuppressive function.Methods 1, Using four-color direct fluorescence-labeled and multi-parameter flow cytometry to detect 40 cases of SLE patients (19 cases of primary and 21 cases of remission) and 15 cases of healthy peripheral blood CD4+ CD25+ T cells and other Treg cells accounted for the proportion of CD4+ T cells. Meanwhile, analysed the correlations among 7 groups and the anti-dsDNA antibodies etc.2, Sorting by flow cytometry combined with cell culture technology to detect and analysis CD4+ CD25+ CD127low / - regulatory T cells to CD4+ CD25-effector T cell proliferation inhibition.Results 1,The 7 groups ratio of SLE patients, respectively: 6.1±1.7; 3.13±1.32; 2.14±1.03; 1.61±0.33; 0.97±0.28; 0.69±0.23 and 0.71±0.35. 2,In SLE group,the proportion of first 6 groups were lower than the control group (P <0.05);For CD4+ CD127low / - Foxp3+ , there was no difference in the two groups(P> 0.05). 3,SLE patients: CD4+ CD25+ FoxP3+, CD4+ CD25high FoxP3+ T cells and IgA ratio was positively correlated respectively; CD4+ CD25high CD127low/-T cells and anti-SSB antibodies was positively correlated.4,SLE patients with early-onset group of seven cells,in addition to CD4+CD127low/-FoxP3+ T cell ratio,the rest were lower than remission(P <0.05).5,The cell proportion of the firt 6 were lower than after hormone treatment (P <0.05).6,In the early onset group,remission group and the control group,CD4+ CD25+ T cells and CD4+ CD25high T cells in Foxp3 expression and low expression of CD127 was positively correlated.7,In SLE patients and control,CD4+ CD25-effect T cells can be suppressed by their own CD4+ CD25+CD127low/- regulatory T cells in vitro,and the inhibition of SLE patients was significantly lower than control.Conclusion SLE may be associated with abnormal immune regulatory T cells and function of the number of defects; CD127 as a possible alternative to Foxp3 regulatory T cell-specific surface markers.The immune abnormalities of SLE may be related to regulatory T cell number and function of defects;CD127 as a possible alternative to Foxp3 regulatory T cell-specific surface marker. Purpose: To investigate the up-regulation mechanism of affecting Treg cells ratio in SLE patients undergone glucocorticoid therapies ,through the research of in vivo and in vitro observing changes of CD4+CD25- T lymphocytes apoptosis in peripheral blood of SLE patients before and after glucocorticoid therapies.Methods: Early stage CD4 +CD25- T lymphocytes apoptosis ration were tested with AnnexinV detection kit and Flow cytometer in 10 cases of SLE patients, who have not had glucocorticoid therapies before and the detection were undertaked with dexamethasone therapies at 0, 1,3 and 7 days. Cell cultivation technology is also adopted to test CD4+CD25- T lymphocytes early apoptosis rate when dexamethasone stimulation are applied to peripheral blood mononuclear cells (PBMC) in vitro for 0, 3 and 7 days.Results: 1.After dexamethasone therapies, SLE patients PBMC CD4+CD25- T lymphocytes early apoposis rate rises obviously as compared with before the treatment(P<0.05).The apoptosis rates were 0.21±0.03,1.39±0.22,1.57±0.11 and 1.83±0.31 at 0, 1, 3 and 7 days after treatment respectively. There was no significant change in early apoptosis rate for CD4+CD25+Treg(P>0.05). The apoptosis rates for 0, 1 day, 3 day and 7 day treatment were 2.02±0.29,2.50±0.59 and 3.72±0.36 repectively.2.After in vitro dexamethasone cultivation of untreated SLE patients' PBMC, CD4+CD25-T lymphocytes early apoptosis rate rose(P<0.05). The apoptosis of vitro cultivation at 1 day,3 day and 7 day were 2.02±0.29,2.50±0.59 and 3.72±0.36 respectively.3.After in vitro dexamethasone cultivation of untreated SLE patients' PBMC,CD4+CD25- T lymphocytes early apoptosis increased with the increase of dexamethasone dose (P <0.05). Cultured in vitro at 7 days, apoptosis rates were 0.60±0.26,1.51±0.69,2.20±0.80 and3.65±0.72 when the corresponding dose of DMX were 0,5,10 and 25 ug .4.Early apoptosis of SLE patients' CD4+CD25- effector T lymphocyte was not correlated to anti-ds-DNA antibodies, anti-Sm antibodies, IgG, IgA, IgM, anti-SSA antibodies, anti-SSB antibodies, C3 and C4, etc (P>0.05), and was negatively related to SLEDAI score(P <0.05).Conclusion: Peripheral blood CD4+CD25- effector T cells'apoptosis of SLE patients treated with dexamethasone increases and is dose-related, leading to peripheral blood CD4+CD25- effector T cells reduction.It is the possible reason for relative increase of CD4+CD25+ Treg cells ratio.