Shimizu Fumitaka, Koga Michiaki, Mizukami Yoichi, Watanabe Kenji, Sato Ryota, Takeshita Yukio, Maeda Toshihiko, Kanda Takashi, Nakamori Masayuki
Department of Neurology and Clinical Neuroscience, Yamaguchi University Graduate School of Medicine, Ube, Japan.
Faculty of Medicine and Health Sciences, Yamaguchi University Graduate School of Medicine, Ube, Japan.
Neurol Neuroimmunol Neuroinflamm. 2025 Jul;12(4):e200405. doi: 10.1212/NXI.0000000000200405. Epub 2025 May 19.
Breakdown of the blood-nerve barrier (BNB) is observed in patients with Guillain-Barré syndrome (GBS); however, the molecular mechanism underlying this phenomenon remains unclear.The aim of this study was to identify antibodies against the BNB-endothelial cells that initiate BNB breakdown in patients with GBS.
We purified IgGs from the serum samples of patients with GBS (n = 77) during the acute phase, disease controls ([DCs], n = 51), and healthy controls ([HCs], n = 24). Human peripheral nerve microvascular endothelial cells (PnMECs) were incubated with IgG. Molecular changes in PnMECs after GBS-IgG exposure were evaluated using RNA-seq and a high-content imaging system. U1-small nuclear ribonucleoprotein (U1-snRNP) autoantibodies were detected using an ELISA. The clinical information of U1-snRNP antibody-positive GBS patients was verified.
GBS-IgGs significantly increased NF-κB nuclear translocation and permeability of the 10-kDa dextran in PnMECs compared with DC-IgGs or HC-IgGs. RNA-seq analyses of PnMECs demonstrated that NF-κB p65 in the center of the network analysis, snRNPs as upstream genes of NF-κB p65, and CXCR5 as downstream genes of NF-κB p65 were important molecules after GBS-IgG exposure. The protein level of claudin-5 and U1-snRNP was significantly reduced while that of CXCR5 was significantly increased after incubation with IgG from patients with GBS, compared with that from HCs. The rate of U1-snRNP antibody positivity was 36% (28 of 77) in patients with GBS, 7% (2 of 28) in DCs, and 0% (0 of 16) in HCs. The serum titer of snRNP antibody decreased after treatment. Both cerebral spinal fluid protein and albumin quotient (QALB)/QALBLIM were higher in snRNP antibody-positive GBS patients than in snRNP antibody-negative GBS patients. IgG from U1-snRNP antibody-positive GBS patients decreased the barrier function and claudin-5 expression more than that from HCs in an in vitro BNB coculture model. The reduction in U1-snRNP antibody decreased the biological effect of IgG from GBS patients with U1-snRNP antibody on the increased permeability of PnMECs.
U1-snRNP autoantibodies are associated with the breakdown of BNB in GBS, through the reduction of U1-snRNP and claudin-5 and the induction of NF-κB activation in BNB-endothelial cells. A temporary autoantibody response against snRNP may be boosted by the periodic response to infection in GBS.
在吉兰-巴雷综合征（GBS）患者中观察到血-神经屏障（BNB）的破坏；然而，这一现象背后的分子机制仍不清楚。本研究的目的是鉴定在GBS患者中引发BNB破坏的针对BNB内皮细胞的抗体。
我们从急性期GBS患者（n = 77）、疾病对照（[DCs]，n = 51）和健康对照（[HCs]，n = 24）的血清样本中纯化免疫球蛋白G（IgG）。将人周围神经微血管内皮细胞（PnMECs）与IgG孵育。使用RNA测序和高内涵成像系统评估GBS-IgG暴露后PnMECs的分子变化。使用酶联免疫吸附测定（ELISA）检测U1-小核糖核蛋白（U1-snRNP）自身抗体。对U1-snRNP抗体阳性的GBS患者的临床信息进行核实。
与DC-IgG或HC-IgG相比，GBS-IgG显著增加了PnMECs中核因子-κB（NF-κB）的核转位以及10 kDa葡聚糖的通透性。对PnMECs的RNA测序分析表明，网络分析中心的NF-κB p65、作为NF-κB p65上游基因的snRNPs以及作为NF-κB p65下游基因的CXC趋化因子受体5（CXCR5）是GBS-IgG暴露后的重要分子。与HCs相比，用GBS患者的IgG孵育后，闭合蛋白-5和U1-snRNP的蛋白水平显著降低，而CXCR5的蛋白水平显著升高。GBS患者中U1-snRNP抗体阳性率为36%（77例中的28例），DCs中为7%（28例中的2例），HCs中为0%（16例中的0例）。治疗后snRNP抗体的血清滴度降低。snRNP抗体阳性的GBS患者的脑脊液蛋白和白蛋白商（QALB）/QALBLIM均高于snRNP抗体阴性的GBS患者。在体外BNB共培养模型中，U1-snRNP抗体阳性的GBS患者的IgG比HCs的IgG更能降低屏障功能和闭合蛋白-5的表达。U1-snRNP抗体的减少降低了U1-snRNP抗体阳性的GBS患者的IgG对PnMECs通透性增加的生物学效应。
U1-snRNP自身抗体通过降低U1-snRNP和闭合蛋白-5以及诱导BNB内皮细胞中的NF-κB激活，与GBS中BNB的破坏有关。针对snRNP的暂时性自身抗体反应可能在GBS中因对感染的周期性反应而增强。
