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1,25-二羟维生素D3处理的树突细胞在变应性气道炎症中的免疫调节作用
2009年
目的探讨1,25-二羟维生素D,[1,25(OH)2D3]处理的树突细胞(DC)在变应性气道炎症中的免疫调节作用及其机制。方法小鼠骨髓来源DC分2组,分别用1,25(OH)2D3和磷酸盐缓冲液(PBS)处理,RT—PCR和蛋白质印迹法检测2组DC Notch配体mRNA和蛋白表达。将2组DC和Notch配体中和抗体封闭的1,25(OH)2D3处理DC分别与小鼠脾脏来源CD4+T细胞共培养,流式细胞仪检测CD4+T细胞中CD4+CD25+Foxp3+T细胞百分比。将卵清蛋白(OVA)致敏小鼠分2组,每组5只,分别过继转移1,25(OH)2D3组DC[1,25(OH)2D3-DC组]和PBS组DC(PBS—DC组),以OVA激发气道炎症后行肺脏病理、支气管肺泡灌洗液(BALF)中白细胞介素4(IL-4)、IL-5、IL-13和干扰素γ(IFN-γ)水平以及脾脏CD4+T细胞中CD4+CD25+Foxp3+T细胞百分比检查。结果1,25(OH)2D3组DCNotch配体Jagged1和Jagged2 mRNA表达(0.3764±0.029、0.5644±0.018)和蛋白表达(0.786±0.034、0.632±0.026)均明显高于PBS组(分别为0.146±0.032、0.267±0.012和0.124±0.025、0.098±0.012,均P〈0.01)。与CD4+T细胞共培养后,1,25(OH)2D3组CD4+T细胞中CD4+CD25+Foxp3+T细胞百分比(22.49%±0.56%)明显高于PBS组(6.67%±0.60%,P〈0.01);经Jagged2中和抗体封闭组CD4+CD25+Foxp3+T细胞百分比(6.56%±1.89%)明显低于未封闭组(20.37%±1.64%,P〈0.01)。1,25(OH)2D3-DC组小鼠气道炎症明显轻于PBS-DC组;BALF中IL-4、IL-5、IL.13和IFN-γ水平(pg/ml)分别为33±5、134±23、91±11和未检测到(〈12.5),均明显低于PBS—DC组(分别为55±7、332±49、152±19、23±6,均P〈0.01);脾脏CD4+T细胞中CD4+CD25+Foxp3+T细胞百分比(14.69%±1.14%)明显高于PBS—DC组(2.38%±0.14%,P〈0.01)。结论1,25(OH)2D3处理DC对变应性气道炎症有抑制
夏俊波王长征马建新安晓静
关键词:骨化三醇树突细胞T淋巴细胞
1,25-Dihydroxyvitamin D3 pretreatment enhances the efficacy of allergen immunotherapy in a mouse allergic asthma model被引量:1
2010年
Background Allergen-specific immunotherapy can induce immune tolerance to specific allergens by regulating immune status of individuals. However, its clinical application is limited due to individual differences in efficacy among patients and un-confirmed safety. 1,25 Dihydroxyvitamin D3 (1,25(OH)2D3) has been shown to be involved in a variety of physiological processes, including immune response regulation. In the present study we explored the role of 1,25(OH)2D3 pretreatment for immunotherapy.Methods Seventy-five BALB/c mice were randomly divided into five groups (15 mice per group). The mouse allergic asthma model was established by intra-peritoneal injection of ovalbumin (OVA, 10 μg) and aluminium hydroxide (2 mg)as an adjuvant. Intra-peritoneal injection of 50 ng of 1,25(OH)2D3 served as a pretreatment, subcutaneous injection of OVA (100 μg) as an immunotherapy, and 1% OVA inhalation as a challenge. Histopathological analysis was performed on four mice per group. The number of cells and their classification in bronchoalvolar lavage (BAL) fluid were assayed.Levels of serum OVA-specific immunoglobulin E (slgE) and IFN-Y, IL-4, IL-5 and IL-10 in BAL fluid were measured by ELISA.Results After 1,25(OH)2D3 pretreatment, immunotherapy could significantly inhibit the infiltration of inflammatory cells into lung tissues and BAL fluid of mice with allergic asthma when compared with un-treated animals (eosinophils:(7.46±1.34)×104/ml vs. (13.41±1.67)×104/ml, P <0.05). In addition, levels of IL-4 ((36.g1±7.87) pg/ml vs. (43.70±6.42)pg/ml, P >0.05) and IL-5 ((41.97±7.93) pg/ml vs. (60.14±8.35) pg/ml, P <0.05) in BAL fluid and serum slgE ((0.42±0.05)vs. (0.75±0.06) OD units, P <0.05) were profoundly reduced. However, the IL-10 level in BAL fluid was significantly increased ((67.74±6.57) pg/ml vs. (44.62±8.81) pg/ml, P <0.05).Conclusions These results indicated that 1,25(OH)2D3 pretreatment e
MA Jian-xin XIA Jun-bo CHENG Xiao-ming WANG Chang-zheng
关键词:ASTHMAIMMUNOTHERAPY
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