Neuronal Toll-like receptor 4 signaling induces brain endothelial activation and neutrophil transmigration in vitro.
Neuronal Toll-like receptor 4 signaling induces brain endothelial activation and neutrophil transmigration in vitro.
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DOI：
10.1186/1742-2094-9-230
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发表时间：
2012-10-03
影响因子：
9.3
通讯作者：
Pinteaux E
中科院分区：
文献类型：
--
作者：
Leow-Dyke S;Allen C;Denes A;Nilsson O;Maysami S;Bowie AG;Rothwell NJ;Pinteaux E
脑中的先天性免疫应答由响应于中枢神经系统（CNS）感染或损伤而产生的病原体相关分子模式（PAMPS）或病原体相关分子模式（DAMPS）启动。这些分子激活Toll样受体（TLR）家族的成员，其中TLR 4是细菌脂多糖（LPS）的受体。虽然已经报道神经元表达TLR 4，但TLR 4在神经元中的激活功能仍然未知。通过RT-PCR评估原代小鼠神经胶质细胞和神经元培养物中TLR 4 mRNA的表达。在不存在或存在TLR 4特异性拮抗剂（VIPER）或特异性JNK抑制剂（SP 600125）的情况下，用LPS处理小鼠混合的神经胶质、神经元或内皮细胞培养物。流式细胞仪微球阵列（CBA）和ELISA法检测炎症介质的表达。Western blot检测细胞外信号调节激酶1/2（ERK 1/2）、p38、c-Jun-N-末端激酶（JNK）和c-Jun的活化。通过中性粒细胞迁移试验评估未经处理的与LPS处理的胶质细胞或神经元培养物的条件培养基对内皮活化的影响，并使用免疫细胞化学和ELISA来测量细胞间细胞粘附分子（ICAM-1）和血管细胞粘附分子（VCAM-1）的表达。在原代小鼠神经元培养物中，LPS诱导趋化因子RANTES和CXCL 1（KC）、肿瘤坏死因子-α（TNFα）和IL-6的强烈释放。相反，LPS诱导混合胶质细胞释放IL-1α、IL-1β和粒细胞集落刺激因子（G-CSF），但在神经元培养物中不诱导。LPS诱导的神经元KC表达和释放被VIPER完全阻断。在神经胶质细胞培养中，LPS诱导ERK 1/2、p38和JNK的活化。相反，在神经元培养中，LPS激活JNK而非ERK 1/2或p38，并且特异性JNK抑制剂SP 600125显著阻断LPS诱导的KC表达和释放。最后，LPS处理的神经元培养物的条件培养基诱导内皮细胞上ICAM-1和VCAM-1的强烈表达，并诱导中性粒细胞浸润穿过内皮单层，这被VIPER抑制。这些数据首次表明，神经元可以作为感染的关键传感器发挥作用，从而引发CNS炎症。
The innate immune response in the brain is initiated by pathogen-associated molecular patterns (PAMPS) or danger-associated molecular patterns (DAMPS) produced in response to central nervous system (CNS) infection or injury. These molecules activate members of the Toll-like receptor (TLR) family, of which TLR4 is the receptor for bacterial lipopolysaccharide (LPS). Although neurons have been reported to express TLR4, the function of TLR4 activation in neurons remains unknown. TLR4 mRNA expression in primary mouse glial and neuronal cultures was assessed by RT-PCR. Mouse mixed glial, neuronal or endothelial cell cultures were treated with LPS in the absence or the presence of a TLR4 specific antagonist (VIPER) or a specific JNK inhibitor (SP600125). Expression of inflammatory mediators was assayed by cytometric bead array (CBA) and ELISA. Activation of extracellular-signal regulated kinase 1/2 (ERK1/2), p38, c-Jun-N-terminal kinase (JNK) and c-Jun was assessed by Western blot. The effect of conditioned media of untreated- versus LPS-treated glial or neuronal cultures on endothelial activation was assessed by neutrophil transmigration assay, and immunocytochemistry and ELISA were used to measure expression of intercellular cell adhesion molecule (ICAM-1) and vascular cell adhesion molecule (VCAM-1). LPS induces strong release of the chemokines RANTES and CXCL1 (KC), tumor necrosis factor-α (TNFα) and IL-6 in primary mouse neuronal cultures. In contrast, LPS induced release of IL-1α, IL-1β and granulocyte-colony stimulating factor (G-CSF) in mixed glial, but not in neuronal cultures. LPS-induced neuronal KC expression and release were completely blocked by VIPER. In glial cultures, LPS induced activation of ERK1/2, p38 and JNK. In contrast, in neuronal cultures, LPS activated JNK but not ERK1/2 or p38, and the specific JNK inhibitor SP600125 significantly blocked LPS-induced KC expression and release. Finally, conditioned medium of LPS-treated neuronal cultures induced strong expression of ICAM-1 and VCAM-1 on endothelial cells, and induced infiltration of neutrophils across the endothelial monolayer, which was inhibited by VIPER. These data demonstrate for the first time that neurons can play a role as key sensors of infection to initiate CNS inflammation.
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影响因子：
15.9
作者：
Okun, Eitan;Griffioen, Kathleen J.;Mattson, Mark P.
通讯作者：
Mattson, Mark P.
影响因子：
3.7
作者：
Frei R;Steinle J;Birchler T;Loeliger S;Roduit C;Steinhoff D;Seibl R;Büchner K;Seger R;Reith W;Lauener RP
通讯作者：
Lauener RP
影响因子：
4.7
作者：
Saini MG;Pinteaux E;Lee B;Bix GJ
通讯作者：
Bix GJ
影响因子：
21.3
作者：
Rolls, Asya;Shechter, Ravid;Schwartz, Michal
通讯作者：
Schwartz, Michal
影响因子：
4.7
作者：
Pinteaux, E;Parker, LC;Luheshi, GN
通讯作者：
Luheshi, GN