Suzukawa Maho, Komiya Akiko, Iikura Motoyasu, Nagase Hiroyuki, Yoshimura-Uchiyama Chitose, Yamada Hirokazu, Kawasaki Hiroshi, Ohta Ken, Matsushima Kouji, Hirai Koichi, Yamamoto Kazuhiko, Yamaguchi Masao
Department of Allergy and Rheumatology, University of Tokyo Graduate School of Medicine, 7-3-1 Hongo, Tokyo 113-8655, Japan.
Int Immunol. 2006 Nov;18(11):1575-83. doi: 10.1093/intimm/dxl090. Epub 2006 Sep 19.
In allergic disorders, basophils migrate from the blood stream to inflamed tissue sites. Since trans-basement membrane migration is an important step for local basophil accumulation, we performed a human basophil transmigration assay using a model basement membrane, Matrigel. IL-3 in the upper chamber was critical for basophil trans-basement membrane migration over baseline levels, since none of the chemoattractants placed in the lower chambers induced migration. RANTES, IL-8, 5-oxo-6E,8Z,11Z,14Z-eicosatetraenoic acid (5-oxo-ETE) and platelet-activating factor (PAF) significantly up-regulated the transmigration of IL-3-treated basophils. Neutralizing experiments indicated the involvement of beta2 integrin and matrix metalloproteinase (MMP)-2/9 in basophil transmigration. Real-time quantitative PCR revealed that basophils constitutively expressed transcripts for MMP-9, and at lower levels, MMP-2, but cell-surface expression was only detected for MMP-9. MMP-9 was also detected in the cytoplasm and culture supernatant of the basophils. Treatment with IL-3 up-regulated the surface level of MMP-9 on the basophils. Our results suggest that basophils possess a unique regulatory mechanism for trans-basement membrane migration which is affected by cytokines, chemoattractants, beta2 integrin and MMPs, especially MMP-9. MMP-9 may be critically involved in the pathogenesis of local basophil influx in allergic diseases.
在过敏性疾病中，嗜碱性粒细胞从血流迁移至炎症组织部位。由于跨基底膜迁移是嗜碱性粒细胞在局部积聚的重要步骤，我们使用基质胶（Matrigel）这一模型基底膜进行了人嗜碱性粒细胞迁移试验。上室中的白细胞介素-3对于嗜碱性粒细胞跨基底膜迁移超过基线水平至关重要，因为置于下室的趋化因子均未诱导迁移。调节激活正常T细胞表达和分泌的因子（RANTES）、白细胞介素-8、5-氧代-6E,8Z,11Z,14Z-二十碳四烯酸（5-氧代-ETE）和血小板活化因子（PAF）显著上调了经白细胞介素-3处理的嗜碱性粒细胞的迁移。中和实验表明β2整合素和基质金属蛋白酶（MMP）-2/9参与了嗜碱性粒细胞迁移。实时定量PCR显示嗜碱性粒细胞组成性表达MMP-9的转录本，且MMP-2的表达水平较低，但仅检测到MMP-9的细胞表面表达。在嗜碱性粒细胞的细胞质和培养上清液中也检测到了MMP-9。白细胞介素-3处理上调了嗜碱性粒细胞表面MMP-9的水平。我们的结果表明，嗜碱性粒细胞具有独特的跨基底膜迁移调节机制，该机制受细胞因子、趋化因子、β2整合素和基质金属蛋白酶影响，尤其是MMP-9。MMP-9可能在过敏性疾病中局部嗜碱性粒细胞流入的发病机制中起关键作用。
