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1,25-(OH)2D3抑制脂多糖诱导的脐血CD4+T细胞IL-13和IL-17的表达
钟慧, 周小建, 洪建国
中国当代儿科杂志 ›› 2013, Vol. 15 ›› Issue (9) : 763-766.
PDF(1330 KB)
PDF(1330 KB)
1,25-(OH)2D3抑制脂多糖诱导的脐血CD4+T细胞IL-13和IL-17的表达
1,25-(OH)2D3 inhibits lipopolysaccharide-induced expression of interleukin-13 and interleukin-17 in cord blood CD4+T cells
目的 本研究旨在探讨生命早期脂多糖(LPS)对脐血CD4+T细胞白介素-13(IL-13)和白介素-17(IL-17)表达的影响及1,25-(OH)2D3对其表达的干预作用,为维生素D的临床合理应用及其对哮喘等变态反应性疾病的防治提供理论依据。方法 选取顺产的足月新生儿12例,断脐后立即取胎盘端脐静脉血50 mL,采用密度梯度离心法分离脐血单个核细胞(CBMCs),磁珠分选CD4+T细胞后依据不同的处理方法分为空白刺激组、LPS(10 μg/mL)单独刺激组和LPS(10 μg/mL)+1,25-(OH)2D3(10-8 mmol/L)共刺激组。培养72 h后采用双抗夹心酶联免疫吸附试验(ELISA)和Real-Time PCR分别检测培养上清液中IL-13和IL-17水平及CD4+T细胞中IL-13和IL-17 mRNA表达。结果 与空白刺激组相比,LPS单独刺激组培养上清液中IL-13和IL-17水平及CD4+T细胞中IL-13和IL-17 mRNA表达水平明显升高(均P<0.01),而1,25-(OH)2D3处理可降低其表达水平(均P<0.05),但仍高于空白刺激组(均P<0.01)。结论 LPS可促进脐血CD4+T细胞IL-13和IL-17的表达;1,25-(OH)2D3对于LPS诱导的脐血CD4+T细胞IL-13和IL-17表达具有抑制作用,提示1,25-(OH)2D3可能在变应原致敏的早期发挥一定保护作用。
Objective To study the effect of 1,25-(OH)2D3 on lipopolysaccharide (LPS)-induced expression of interleukin-13 (IL-13) and interleukin-17 (IL-17) in cord blood CD4+T cells, providing theoretical basis for clinical reasonable application of vitamin D and prevention of asthma and allergic diseases. Methods Mononuclear cells (MNCs) were isolated from umblilical cord blood (50 mL) of 12 normal eutocia term newborns by gravity centrifugation. CD4+T cells were isolated using magnetic beads, which was cultured with following three kinds of stimulus for 72 hours:. natural state (blank group), LPS (10 μg/mL)stimulation alone and LPS(10 μg/mL)+1,25-(OH)2D3 (10-8 mmol/L)stimulation. Levels of IL-13 and IL-17 in the culture supernatant and mRNA expressions in cord blood CD4+T cells were detected using ELISA and real Time-PCR respectively. Results Compared with the blank group, levels of IL-13 and IL-17 in the culture supernatant and mRNA expression of IL-13 and IL-17 in the cord blood CD4+T cells increased in the LPS stimulation alone group (P<0.01). When co-stimulation of 1,25-(OH)2D3 with LPS, levels of IL-13 and IL-17 in the culture supernatant and mRNA expression of IL-13 and IL-17 in the cord blood CD4+T cells decreased compared with LPS-stimulated alone group (P<0.05), but remained higher than the blank group (P<0.01). Conclusions LPS can promote expression of IL-13 and IL-17 in cord blood CD4+T cells. 1,25-(OH)2D3 inhibits the expression of IL-13 and IL-17, suggesting that vitamin D intake may provide protective effects in the development of atopy-predisposing immune responses in early life.
脂多糖 / 1 / 25-(OH)2D3 / 白介素-13 / 白介素-17 / CD4+T细胞
Lipopolysaccharide / 1,25-(OH)2D3 / Interleukin-13 / Interleukin-17 / CD4+T cell
[1] Thornton CA, Upham JW, Wikstrom ME, Holt BJ, White GP, Sharp MJ, et al. Functional maturation of CD4+CD25+CTLA4+CD45RA+ T regulatory cells in human neonatal T cell responses to environmental antigens/allergens[J]. J Immunol, 2004, 173(5): 3084-3092.
[2] Williams TJ, Jones CA, Miles EA, Warner JO, Warner JA. Fetal and neonatal IL-13 production during pregnancy and at birth and subsequent development of atopic symptoms[J]. J Allergy Clin Immunol, 2000, 105(5): 951-959.
[3] Smillie FI, Elderfield AJ, Patel F, Cain G, Tavenier G, Brutsche M, et al. Lymphoproliferative responses in cord blood and at one year: no evidence for the effect of in utero exposure to dust mite allergens[J]. Clin Exp Allergy, 2001, 31(8): 1194-1204.
[4] Devereux G, Seaton A, Barker RN. In utero priming of allergen-specific helper T cells[J]. Clin Exp Allergy, 2001, 31(11): 1686-1695.
[5] Ko SK, Jin M, Pyo MY. Inonotus obliquus extracts suppress antigen-specific IgE production through the modulation of Th1/Th2 cytokines in ovalbumin-sensitized mice[J]. J Ethnopharmacol, 2011, 137(3): 1077-1082.
[6] McKenzie AN, Culpepper JA, de Waal Malefyt R, Briere F, Punnonen J, Aversa G, et al. Interleukin 13, a T-cell-derived cytokine that regulates human monocyte and B-cell function[J]. Proc Natl Acad Sci USA, 1993, 90(8): 3735-3739.
[7] Roussel L, Houle F, Chan C, Yao Y, Berube J, Olivenstein R, et al. IL-17 promotes p38 MAPK-dependent endothelial activation enhancing neutrophil recruitment to sites of inflammation[J]. J Immunol, 2010, 184(8): 4531-4537.
[8] Pfeffer PE, Hawrylowicz CM. Vitamin D and lung disease[J]. Thorax, 2012, 67(11): 1018-1020.
[9] Bener A, Ehlayel MS, Tulic MK, Hamid Q. Vitamin D deficiency as a strong predictor of asthma in children[J]. Int Arch Allergy Immunol, 2011, 157(2): 168-175.
[10] Carroll KN, Gebretsadik T, Larkin EK, Dupont WD, Liu Z, Van Driest S, et al. Relationship of maternal vitamin D level with maternal and infant respiratoy disease[J]. Am J Obstet Gynecol, 2011, 205(3): 215e1-215e7.
[11] Morales E, Romieu I, Guerra S, Ballester F, Rebagliato M, Vioque J, et al. Maternal vitamin D status in pregnancy and risk of lower respiratory tract infections,wheezing, and asthma in offspring.Epidemiology[J], 2011, 23(1): 64-71.
[12] Harvey L, Burne TH, McGrath JJ, Eyles DW. Developmental vitamin D3 deficiency induces alterations in immune organ morphology and function in adult offspring[J]. J Steroid Biochem Mol Biol, 2010, 121(1-2): 239-242.
[13] Borges MC, Martini LA, Rogero MM. Current perspectives on vitamin D, immune system, and chronic diseases[J]. Nutrition, 2011, 27(4): 399-404.
[14] Ingram JL, Kraft M. IL-13 in asthma and allergic disease: Asthma phenotypes and targeted therapies[J]. J Allergy Clin Immunol, 2012, 130(4): 829-842.
[15] Baeke F, Korf H, Overbergh L, van Etten E, Verstuyf A, Gysemans C, et al. Human T lymphocytes are direct targets of 1,25-dihydroxyvitamin D3 in the immune system[J]. J Steroid Biochem Mol Biol, 2010, 121(1-2): 221-227.
[16] Topilski I, Flaishon L, Naveh Y, Harmelin A, Levo Y, Shachar I. The anti-inflammatory effects of 1,25-dihydroxyvitamin D3 on Th2 cells in vivo are due in part to the control of integrin-mediated T lymphocyte homing[J]. Eur J Immunol, 2004, 34(4): 1068-1076.
[17] Sloka S, Silva C, Wang J, Yong VW. Predominance of Th2 polarization by vitamin D through a STAT6-dependent mechanism[J]. J Neuroinflammation, 2011, 8(1): 56-66.
[18] Matheu V, Back O, Mondoc E, Issazadeh-Navikas S. Dual effects of vitamin D-induced alteration of TH1/TH2 cytokine expression: enhancing IgE production and decreasing airway eosinophilia in murine allergic airway disease[J]. J Allergy Clin Immunol, 2003, 112(3): 585-592.
[19] Zhao CQ, Li TL, He SH, Chen X, An YF, Wu WK, et al. Specific immunotherapy suppresses Th2 responses via modulating TIM1/TIM4 interaction on dendritic cells[J]. Allergy, 2010, 65(8): 986-995.
[20] 栾斌, 王亚哲, 张艳丽, 谷惠茹, 李彦玲, 赵杰. 1,25-(OH)2D3对哮喘小鼠肺内TIM4表达的影响[J]. 中国当代儿科杂志, 2013, 15(1): 67-70.
[21] Bullens DM, Truyen E, Coteur L, Dilissen E, Hellings PW, Dupont LJ, et al. IL-17 mRNA in sputum of asthmatic patients: linking T cell driven inflammation and granulocytic influx?[J]. Respir Res, 2006, 7(1): 135-142.
[22] McKinley L, Alcorn JF, Peterson A, Dupont RB, Kapadia S, Logar A, et al. TH17 cells mediate steroid-resistant airway inflammation and airway hyperresponsiveness in mice[J]. J Immunol, 2008, 181(6): 4089-4097.
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