寻常性银屑病患者外周血CD4^＋CD25^＋调节性T细胞的功能研究  

A study on the function of CD4^＋CD25^＋ regulatory T cells in peripheral blood of patients with psoriasis vulgaris

作　　者：黄琼[1] 韩凌[1] 方栩[1] 郑志忠[1] 

机构地区：[1]复旦大学附属华山医院皮肤科,上海200040

出　　处：《中华皮肤科杂志》2010年第4期251-255,共5页Chinese Journal of Dermatology

基　　金：国家自然科学基金（30600542）

摘　　要：目的测定银屑病患者外周血中的CD4^＋CD25^＋调节性T细胞功能的改变，探讨调节性T细胞在其发病中的作用。方法寻常性银屑病患者12例，银屑病面积和严重度指数（PASI）评分15～34．5分，均为慢性斑块状。健康对照组6例，为健康献血者。通过免疫磁珠体外分离外周血中的CD4’CD25’T细胞和CD4＋CD25一T细胞，纯度达90％以上。采用[^3H]-脱氧胸腺嘧啶苷方法检测CD4^＋CD25^＋T细胞的增殖和抑制功能，ELISA法测定细胞因子和RT—PCR检测Foxp3mRNA的表达。结果多克隆刺激后CD4＋CD25＋T细胞无明显增殖，IL-2可逆转此无反应状态。健康对照组CD4^＋CD25^＋T细胞CD4^＋CD25^-T细胞增殖的抑制率可达60％，而银屑病组抑制率仅为19．5％（P〈O．01）。银屑病患者CD4^＋CD25^-T细胞单独培养及与CD4^＋CD25^＋T细胞共培养后，分泌IFN-γ水平分别为（179．66±48．97）ng／L和（109．55±48．04）ng／L，健康对照组分别为（87．36±33．36）ng／L和（32．55±15．69）ng/L，两组比较，P值均〈0．05；对IFN-γ分泌的抑制率银屑病患者组为40％，健康对照组为63％（P〈0．05）；健康对照组CD4^＋CD25^＋T细胞和CD4^＋CD25^- T细胞单独培养，转化生长因子-β（TGF—β）分泌分别为（3025±769）ng／L和（2450±431）ng／L（P〈0．05）。两组之间，无论单独培养或两种细胞共培养，IL-10、TGF—β的分泌差异均无统计学意义（P〉0．05）。CD4^＋CD25^＋T细胞表达高水平的Foxp3，银屑病患者和健康人对照调节性T细胞和效应T细胞的Foxp3表达相似。结论银屑病患者外周血中调节性T细胞抑制功能的减弱导致效应T细胞增殖失控，可能参与了银屑病的发病。Objective To analyse the function of CD4^＋CD25^＋ regulatory T cells in patients with psoriasis vulgaris （PV）, and try to explore their role in the pathogenesis of PV. Methods Twelve patients with chronic plaque PV, who had a psoriasis area and severity score （PASI） of 15 to 34.5, were included in this study together with 6 healthy human controls. By using immunomaguetic heads, CD4^＋CD25^＋ regulatory T （Treg） cells and CD4^＋CD25^- responder T （Tresp） cells were isolated from peripheral blood of these subjects. Then, these Treg and Tresp cells were subjected to monoculture or coculture under the stimulation with CD3 and CD28. IL-2 was also used to stimulate the proliferation of the Treg cells. [^3H]-thymidine incorporation assay was performed to analyze the proliferation of cells, enzyme linked immunosorbent assay （ELISA） to detect the levels of interferon-gamma （IFN-γ）, interleukin-10 （IL-10） and transforming growth factor-beta （TGF-β） in the supernatant of monoeuhure or coeulture system, and RT-PCR to measure the expression of Foxp3 mRNA. Results After stimulation with CD3 and CD28, neither normal nor psoriatic Treg cells showed obvious proliferation, while this condition was reversed by the addition of IL-2. Cocuhured with Tresp cells at a ratio of 1：1, Treg cells inhibited the proliferation of Tresp cells by 60% in the control group and 19.5% in the psoriasis group （P 〈 0.01 ）. The level of supernatant IFN-γ was 179.66 ± 48.97 ng/L and 109.55 ± 48.0 ng/L in the monocuhure and coculture system of psoriatic Tresp cells, respectively, significantly different from that of control Tresp cells （87.36 ± 33.36 ng/L, 32.55 ± 15.69 ng/L, both P 〈 0.05）; the secretion of IFN-γ by Tresp cells was suppressed by 40% in psoriatic group and 63% in control group, by Treg cells （P 〈 0.05）. Treg cells secreted more TGF-I3 than Tresp cells in the normal control group （3025 ± 769 ng/L vs 2450 ± 431 ng/L, P 〈 0.05）. Between the patient and control group, t

关 键 词：银屑病 CD4^+CD25^+调节T细胞 CD4^+CD25^-效应T细胞 FOXP3 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]