淋巴毒素-β 受体阻断可减少 NOD 干燥综合征模型中泪腺的 CXCL13 并改善角膜完整性。
Lymphotoxin-beta receptor blockade reduces CXCL13 in lacrimal glands and improves corneal integrity in the NOD model of Sjögren's syndrome.
机构信息
Immunology Research Department, Department of Veterans Affairs Medical Center, 215 North Main Street, White River Junction, VT 05009, USA.
出版信息
Arthritis Res Ther. 2011;13(6):R182. doi: 10.1186/ar3507. Epub 2011 Nov 1.
INTRODUCTION
In Sjögren's syndrome, keratoconjunctivitis sicca (dry eye) is associated with infiltration of lacrimal glands by leukocytes and consequent losses of tear-fluid production and the integrity of the ocular surface. We investigated the effect of blockade of the lymphotoxin-beta receptor (LTBR) pathway on lacrimal-gland pathology in the NOD mouse model of Sjögren's syndrome.
METHODS
Male NOD mice were treated for up to ten weeks with an antagonist, LTBR-Ig, or control mouse antibody MOPC-21. Extra-orbital lacrimal glands were analyzed by immunohistochemistry for high endothelial venules (HEV), by Affymetrix gene-array analysis and real-time PCR for differential gene expression, and by ELISA for CXCL13 protein. Leukocytes from lacrimal glands were analyzed by flow-cytometry. Tear-fluid secretion-rates were measured and the integrity of the ocular surface was scored using slit-lamp microscopy and fluorescein isothiocyanate (FITC) staining. The chemokine CXCL13 was measured by ELISA in sera from Sjögren's syndrome patients (n = 27) and healthy controls (n = 30). Statistical analysis was by the two-tailed, unpaired T-test, or the Mann-Whitney-test for ocular integrity scores.
RESULTS
LTBR blockade for eight weeks reduced B-cell accumulation (approximately 5-fold), eliminated HEV in lacrimal glands, and reduced the entry rate of lymphocytes into lacrimal glands. Affymetrix-chip analysis revealed numerous changes in mRNA expression due to LTBR blockade, including reduction of homeostatic chemokine expression. The reduction of CXCL13, CCL21, CCL19 mRNA and the HEV-associated gene GLYCAM-1 was confirmed by PCR analysis. CXCL13 protein increased with disease progression in lacrimal-gland homogenates, but after LTBR blockade for 8 weeks, CXCL13 was reduced approximately 6-fold to 8.4 pg/mg (+/- 2.7) from 51 pg/mg (+/-5.3) in lacrimal glands of 16 week old control mice. Mice given LTBR blockade exhibited an approximately two-fold greater tear-fluid secretion than control mice (P = 0.001), and had a significantly improved ocular surface integrity score (P = 0.005). The mean CXCL13 concentration in sera from Sjögren's patients (n = 27) was 170 pg/ml, compared to 92.0 pg/ml for sera from (n = 30) healthy controls (P = 0.01).
CONCLUSIONS
Blockade of LTBR pathways may have therapeutic potential for treatment of Sjögren's syndrome.
简介
在干燥综合征中，干眼症与泪腺白细胞浸润以及泪液产生和眼表完整性丧失有关。我们研究了阻断淋巴毒素-β受体（LTBR）途径对干燥综合征 NOD 小鼠模型中泪腺病理学的影响。
方法
雄性 NOD 小鼠接受长达十周的 LTBR-Ig 拮抗剂或对照鼠抗体 MOPC-21 治疗。通过免疫组织化学分析外眶泪腺的高内皮静脉（HEV），通过 Affymetrix 基因芯片分析和实时 PCR 分析差异基因表达，通过 ELISA 分析 CXCL13 蛋白。通过流式细胞术分析泪腺中的白细胞。通过裂隙灯显微镜和荧光素异硫氰酸酯（FITC）染色测量泪液分泌率，并评估眼表完整性评分。通过 ELISA 测量干燥综合征患者（n=27）和健康对照者（n=30）血清中的趋化因子 CXCL13。统计分析采用双尾非配对 T 检验或 Mann-Whitney 检验评估眼表完整性评分。
结果
LTBR 阻断治疗 8 周后，B 细胞累积减少（约 5 倍），HEV 在泪腺中消失，并减少了淋巴细胞进入泪腺的速度。Affymetrix 芯片分析显示，LTBR 阻断后，由于多种 mRNA 表达发生变化，包括稳态趋化因子表达减少。PCR 分析证实，CXCL13、CCL21、CCL19mRNA 和 HEV 相关基因 GLYCAM-1 的减少。CXCL13 蛋白在泪腺匀浆中随疾病进展而增加，但在用 LTBR 阻断治疗 8 周后，与 16 周龄对照小鼠的 51pg/mg（+/-5.3）相比，CXCL13 减少了约 6 倍，至 8.4pg/mg（+/-2.7）。给予 LTBR 阻断的小鼠的泪液分泌量比对照组增加了约两倍（P=0.001），眼表完整性评分显著改善（P=0.005）。干燥综合征患者（n=27）血清中的 CXCL13 平均浓度为 170pg/ml，而健康对照组（n=30）血清中的 CXCL13 浓度为 92.0pg/ml（P=0.01）。
结论
阻断 LTBR 途径可能具有治疗干燥综合征的治疗潜力。
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