The Change Of CD4~+CD25~+Foxp3~+T Cells From Vitiligo Patients In The Autologous Epidermal Graft Treatment Combination With NB-UVB Therapy And The Observation Of Skin CLSM

Background Vitiligo is a common pigmentary disorder, with a incidence rate of 0.5% to 2% all around the world. It presents as local or universal leukoplakia, breaks down the cosmetic appearance, and always leads to sever mental stress. The exact etiological factor of vitiligo is still unclear. There are many different etiology and the immune factor is paid more and more attention. CD4+CD25+Treg was the focus of the research. There are a lot of studies hinted that the abnormal number and function of CD4+CD25+Treg could lead to autoimmune disease. The method to treat vitiligo was difficult, all the nonsurgical method should use long time, which could cause adverse effect, such as the psoralen and their derivate could cause flare and blister. The glucocorticosteroid easily cause atrophia. Autologous epidermal grafting is an easy, safe and effective option for the treatment of stable refractory vitiligo, although which had many factors influence the effect. It is easily performed , safe and effective method of treating vitiligo and the long-term effect is good, with no obviously adverse effect. The mechanism of UVB biology was to irritate the melanocyte to produce more melanin and migration. The autologous epidermal graft combination with the NB-UVB can increase the therapy effects. Confocal laser scanning microscope was a skill based on the opotics copolymerization principle localization, in situ, real-time and dynamic state technolonies to the clincial examination, which had most clinical application non-invasive skin image technolonies recently. Objective: to investigate whether CD4+CD25+ Foxp3+ regulator T cells is related with NB-UVB in the epidermal autologous treatment of vitiligo, Analyzing the relationship between the effect and the change. Using CLSM to observe the leukoplakia of the transplanted area and normal skin before and after treatment, to observe the melain content of skin.Object and method: 43 patients, who had visited out-patient clinic of dermatology of the Third People's Hospital of Hangzhou during September 2008 and Jan 2009, with vitiligo and had accepted autologous epidermal grafting, were recruited for the study. In the tested group, leukoplakia had been irradiated by NB-UVB (10 times) after grafting, and then with another grafting, while the control group had not been treated with NB-UVB. Peripheral blood were collected from the treated group and 19 people from health examination. Flow cytometry was used to analyze the proportion of CD4+CD25+ Foxp3+ in the peripheral blood mononuclear cells (PBMC). Using the digital cameral to record the pictures of the white maculae before and after therapy. Confocal microscopy laser was used to observe the melain of the skin before and after NB-UVB treatment. Data from the study were analyzed withχ2 test, t test and Rank test by SPSS(16.0) to determine the relationship between therapeutic effect of autologous epidermal grafting combination with NB-UVB and Treg cells.Results:①The NB-UVB irradiation can obviously increase the rate of excellent repigmentatin of leukoplakia, and total effective rate. The effect(89.92%) of patients treated with NB-UVB was a little lower than the second effect of graft(95.52%), which had no difference with the untreated group.(χ2=3.07,p>0.05,and 11.12,p<0.05 respectively). The effective of patients(82.98%) treated only with graft was lower than effect of the second graft of combination group(χ2=2.75,p>0.05).②After treatment, the proportion of CD4+CD25+Foxp3+ was significantly higher than that in the untreate group(p<0.05), which had no diffierence with the proportion of control group(p>0.05).③There leukoplakia of 15 patients around the transplanted was more dark after 3 times irridiation. The confocal laser scanning microscopy can show the melain was the same as the normal.Conclusions: The changes of CD4+CD25+ Foxp3+ Treg cell in number might participate in the onset of vitligo, the NB-UVB can increase CD4+CD25+ Foxp3+ Treg frequency in treating vitiligo. NB-UVB combination with autologous graft can increase the effective of repigmentation and improve the beautiful outlook of the graft. The confocal laser scanning microscopy might directly observe the melain of the melanocyte.