Ceramide-CD300f binding suppresses experimental colitis by inhibiting ATP-mediated mast cell activation.
Ceramide-CD300f binding suppresses experimental colitis by inhibiting ATP-mediated mast cell activation.
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DOI：
10.1136/gutjnl-2014-308900
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发表时间：
2016-05
期刊：
影响因子：
24.5
通讯作者：
Kitaura J
中科院分区：
文献类型：
--
作者：
Matsukawa T;Izawa K;Isobe M;Takahashi M;Maehara A;Yamanishi Y;Kaitani A;Okumura K;Teshima T;Kitamura T;Kitaura J
细胞外ATP通过P2 X7嘌呤受体介导肥大细胞依赖性肠道炎症我们之前已经证明，CD 300 f（也称为白细胞单免疫球蛋白样受体3（LMIR 3））通过与神经酰胺结合抑制免疫球蛋白E依赖性和肥大细胞依赖性过敏反应。本研究的目的是阐明神经酰胺-LMIR 3相互作用在IBD发展中的作用。葡聚糖硫酸钠（DSS）诱导的结肠炎模型用于移植有WT或LMIR 3 −/−骨髓源性肥大细胞（BMMC）的野生型（WT）、LMIR 3 −/−、肥大细胞缺陷型KitW-sh/W-sh、KitW-sh/W-shLMIR 3 −/−或KitW-sh/W-sh小鼠。结肠炎的严重程度由临床和组织学标准确定。通过流式细胞术评估固有层细胞群。通过实时逆转录PCR测量固有层细胞的化学介质的产生。用ELISA法测定神经酰胺存在或不存在时ATP刺激的BMMC产生的化学介质。在给予含有LMIR 3的细胞外结构域和抗神经酰胺抗体的Fc融合蛋白或神经酰胺脂质体的小鼠中评估DSS诱导的结肠炎的严重程度。LMIR 3缺乏加重了DSS诱导的小鼠结肠炎。携带LMIR 3 −/−肥大细胞的KitW-sh/W-sh小鼠比携带WT肥大细胞的小鼠表现出更严重的结肠炎。神经酰胺-LMIR 3相互作用抑制ATP刺激的BMMC活化。DSS诱导的结肠炎通过破坏神经酰胺-LMIR 3相互作用而加重，而通过用神经酰胺脂质体治疗而抑制。LMIR 3缺陷型结肠肥大细胞在DSS诱导的LMIR 3 −/−小鼠结肠炎恶化中起关键作用。神经酰胺脂质体通过与LMIR 3结合抑制ATP介导的结肠肥大细胞活化，从而减轻DSS诱导的结肠炎。
Extracellular ATP mediates mast cell-dependent intestinal inflammation via P2X7 purinoceptors. We have previously shown that CD300f (also called the leucocyte mono-immunoglobulin-like receptor 3 (LMIR3)) suppresses immunoglobulin E-dependent and mast cell-dependent allergic responses by binding to ceramide. The aim of the present study was to clarify the role of ceramide–LMIR3 interaction in the development of IBD. The dextran sodium sulfate (DSS)-induced colitis model was used in wild-type (WT), LMIR3−/−, mast cell-deficient KitW-sh/W-sh, KitW-sh/W-shLMIR3−/− or KitW-sh/W-sh mice engrafted with WT or LMIR3−/− bone marrow-derived mast cells (BMMCs). The severity of colitis was determined by clinical and histological criteria. Lamina propria cell populations were assessed by flow cytometry. Production of chemical mediators from lamina propria cells was measured by real-time reverse transcription PCR. Production of chemical mediators from ATP-stimulated BMMCs in the presence or absence of ceramide was measured by ELISA. The severity of DSS-induced colitis was assessed in mice given either an Fc fusion protein containing an extracellular domain of LMIR3, and anticeramide antibody, or ceramide liposomes. LMIR3 deficiency exacerbated DSS-induced colitis in mice. KitW-sh/W-sh mice harbouring LMIR3−/− mast cells exhibited more severe colitis than those harbouring WT mast cells. Ceramide–LMIR3 interaction inhibited ATP-stimulated activation of BMMCs. DSS-induced colitis was aggravated by disrupting the ceramide–LMIR3 interaction, whereas it was suppressed by treating with ceramide liposomes. LMIR3-deficient colonic mast cells were pivotal in the exacerbation of DSS-induced colitis in LMIR3−/− mice. Ceramide liposomes attenuated DSS-induced colitis by inhibiting ATP-mediated activation of colonic mast cells through ceraimide–LMIR3 binding.