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孓瞻寇
5 years ago
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1 CD + 4 CD + 25 T The role of CD + 4 CD + 25 Regulatory T cells in the pathogenesis of vitiligo

3 CD + 4 CD + 25 T The role of CD + 4 CD + 25 Regulatory T cells in the pathogenesis of vitiligo

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6 CD + 4 CD + 25 CD + 4 CD + 25 Vitiligo patients II

7 8+T DETACHaBEAD CD8+T Miltenyi CD4+T CD25 CD4+T CD4+CD25+T CD4+CD25-T 3 CD4+CD25+ T 4 CD4+CD25+ T 5 RNA RT-PCR Foxp3CTLA-4 GITR Western-blot Foxp3 6 SPSS11.0 ± P< CD4+CD25+ T CD4+CD25+ T CD4+CD25+ T NB-UVB 3 CD4+CD25+ T CD4+CD25+ Treg CD4+CD25+ T CD CD 25 CD CD 25 CD CD 25 III

8 CD CD 25 CD CD 25 CD CD 25 IV

9 The role of CD + 4 CD + 25 Regulatory T cells in the pathogenesis of vitiligo NameWu Yan-Hong SupervisorYang Hui-lan ABSTRACT Vitiligo is a common dermatological disorder characterized by milky-whi te depigmented macules devoid of identiiable melanocytes. In the recent year s there is a gradual increase in the incidence of vitiligo. The cause of vitilig o is probably associated with genetic and environmental factors. However, t he exact cause of the illness remains unknown, but several hypotheses about i ts pathogenesis are advanced. The hypothesis of autoimmunity is subjected to more and more concern recently. An exciting new area of immunologic interest has been the identification of immune-regulatory cells (Tregs) and their role in maintaining immune tole rance. The CD 4 + Tregs come from thymus gland, accounting for 5-10% of t he CD + 4 T cells in the peripheral blood. Naturally occurring CD + 4 Tregs, the majority of which express CD 25, can suppress the activation, proliferation an d differentiation of self-activated T cells. Foxp3a transcriptional factor repor ted recently, is a member of the forkhead family of transcriptional regulators. Foxp3 constitute expresses in CD + 4 CD + 25 Tregs and mainly in thymus gland, spleen and lymph nodes. CD + 4 CD + 25 Tregs have two major features, includ ing immunotolerance and immunosuppression. They are tolerant to the stimul ation of the high concentration of IL-2, solid-phase coating or soluble anti-cd i

10 ABSTRACT 3 antibody, and the joint stateof anti-cd3 and anti-cd28 antibodies with no s ecretion of IL-2. The immunosuppression of Tregs shows that the activation of Tregs requires the stimulation of T-cell receptors and some other signals. Once activated, their suppression is not specific. They can cause local immu nosuppression by direct interaction between cells and the secretion of IL-10 a nd TGF-band this immunosuppression has no restriction to MHC. After the activation of Tregs, the expression of CTLA-4 is increased. The anti-ctla-4 antibody can be used to block the immunosuppression of Tregs and it is gen erally believed that Tregs play effects by cell interactions. The glucocorticoid -induced tumor necrosis factor receptor(gitr), the 18 th member of TNFRSF, i s expressed on the surface of Tregs, with the ligand GITRL. GITR and CTL A-4 can increase the expression of Foxp3. The discovery of Foxp3 as a spe cific marker of Tregs has led to an explosion of research in the function of Tregs. Although the role of Tregs in the pathogenesis of vitiligo was reporte d overseas, their results are inconsistent. However, there is no report of this f ield in China yet. Objectives: To study the role of CD + 4 CD + 25 Regulatory T cells in the pathogenesis of vitiligo Methods: 1. The blood was taken from vitiligo patients and normal control and density centrifugation was used to separate the PBMC. 2. CD + 8 T cells were separated by Dynabeads, and Militenyi immunity beads were applied to separate the CD + 4 CD + 25 T cells and CD + 4 CD - 25 T cells. 3. The number of CD + 4 CD + 25 Tregs in PBMC of vitiligo patients and the normal control was analyzed by flow cytometry. 4. Flow cytometry was applied to analyze the number of CD + 4 CD + 25 Tregs in ii

11 PBMC of vitiligo patients both in active stage and post-treatment by NB-UVB. 5. RT-PCR was used to analyze the mrna expression level of Foxp3, CTLA-4 and GITR,and Western-blot was applied to analyze the protein expression level of Foxp3. 6. Statistics analysis: The program SPSS11.0 was used to analyze all the statistical data. When P<0.05, the difference is significant. Results: 1. The number of CD 4 + CD 25 + Tregs in peripheral blood mononuclear cells(pbmc) of 39 vitiligo patients and normal control was analyzed by flow cytometry. The results showed that the percentage of CD + 4 CD + 25 Tregs in PBMC of vitiligo patients in active stage was lower than that of normal control, and the difference is significant. After treated by NB-UVB, the percentage of CD CD 25 Tregs of vitiligo patients was higher than that of vitiligo patients in the active stage and the difference is also significant. These results suggested that the CD CD 25 Regulatory T cells might play an important role in the pathogenesis of vitiligo. After the treatment, the percentage of CD + 4 CD + 25 Tregs was lower than the normal control, but the difference is not significant. 2. The mrna expression levels of Foxp3, CTLA-4 and GITR were analyzed by RT-PCR, and the protein expression level of Foxp3 was analyzed by Western blotting. The results showed that the expression levels of Foxp3, CTLA-4 and GITR in the patients of the active stage were lower than those of the normal control, and the difference is significant. After treated by NB-UVB, the expression level of Foxp3 of the vitiligo patients was lower than that of normal control, while difference of the number of CD + 4 CD + 25 Regulatory T cells between the two groups is not significant. These results demonstrated that after the treatment, the number of CD CD 25 iii

12 ABSTRACT Regulatory T cells had been normalized but their function has possibly been changed. Conclusions: To investigate the role of CD + 4 CD + 25 Tregs in the pathogenesis of vitiligo, this study indicated that after the NB-UVB treatment, the number of CD + 4 CD + 25 Tregs of vitiligo patients had been restored but the expression level of Foxp3 was lower than that of normal control. It suggested that the function CD + 4 CD + 25 Tregs had possibly been changed after the treatment, and the CD + 4 CD + 25 Regulatory T cells might play an important role in the pathogenesis of vitiligo. Key wordsvitiligo, CD 4 + CD 25 + Regulatory T cells, Foxp3, flow cytometry, RT-PCR, Western blot iv

13 ABSTRACT HLA

600 现代药物与临床 Drugs & Clinic 第 31 卷第 5 期 2016 年 5 月 were significantly decreased, but the levels of IL-12 in two groups were significantly increa

600 现代药物与临床 Drugs & Clinic 第 31 卷第 5 期 2016 年 5 月 were significantly decreased, but the levels of IL-12 in two groups were significantly increa 现代药物与临床 Drugs & Clinic 第 31 卷第 5 期 2016 年 5 月 599 喘可治注射液联合细辛脑注射液治疗小儿毛细支气管炎的临床研究唐辉英, 佘定平, 马颐娇深圳市儿童医院呼吸科, 广东深圳 518026 摘要 : 目的探讨喘可治注射液联合