猪链球菌2型胶原酶样蛋白酶增强血脑屏障通透性
促进脑膜炎发生

doi:10.1016/j.jia.2024.06.005

Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (11): 4379-4394.DOI: 10.1016/j.jia.2024.06.005

猪链球菌2型胶原酶样蛋白酶增强血脑屏障通透性促进脑膜炎发生

收稿日期:2023-10-17 修回日期:2024-06-27 接受日期:2024-04-22 出版日期:2025-11-20 发布日期:2025-10-17

Streptococcus suis serotype 2 collagenase-like protease promotes meningitis by increasing blood–brain barrier permeability

Jikun Mei^1*, Xuan Jiang^1*, Fengyang Li¹, Zengshuai Wu¹, Tong Wu¹, Junhui Zhu¹, Hexiang Jiang¹, Ziheng Li¹, Na Li^1#, Liancheng Lei^{1, 2#}

¹State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases/Key Laboratory for Zoonosis Research of Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, China
²College of Animal Science, Yangtze University, Jingzhou 434023, China

Received:2023-10-17 Revised:2024-06-27 Accepted:2024-04-22 Online:2025-11-20 Published:2025-10-17
About author:Jikun Mei, E-mail: mjk20@163.com; Xuan Jiang, E-mail: 18522075540@163.com; #Correspondence Na Li, E-mails: vetlina2013@126.com; Liancheng Lei, E-mails: leiliancheng@163.com *These authors contributed equally to this study.
Supported by:
This study was supported by the National Key Research and Development Program of China (2021FYD1800405) and the National Natural Science Foundation of China (32072823).

摘要/Abstract

摘要：

猪链球菌2型（Streptococcus suis serotype 2，SS2）是一种新兴的人兽共患病原体，可引起人和猪的脑膜炎，不仅给养猪业带来巨大经济损失，还严重威胁公共卫生安全。然而，SS2进入大脑并诱发脑膜炎的机制尚不完全清楚。在这里，我们研究了SS2胶原酶样蛋白酶（collagenase-like protease，Clp）破坏血脑屏障（Blood-Brain Barrier，BBB）促进脑膜炎发生的作用和机制。我们以纯化的SS2 rClp重组蛋白及clp基因缺失菌株为工具，通过小鼠及体外hCMEC/D3单层BBB模型进行相关实验。发现Clp可增强SS2的毒力作用及其组织定植能力，并可促进SS2对小鼠BBB的破坏作用。与野生型SS2相比，Δclp突变体穿过人脑微血管内皮系（hCMEC/D3）单层屏障的能力降低，而重组蛋白rClp的添加增加了通透性。rClp还显著促进SS2对hCMEC/D3的粘附，抑制细胞间紧密连接蛋白ZO-1、Occludin和Claudin-5的表达（独立于其酶活性），并通过细胞受体配体凋亡和线粒体凋亡途径诱导hCMEC/D3凋亡，导致BBB破坏和通透性增加。此外，Clp增加了SS2诱导巨噬细胞（F4/80+）、单核细胞（F4/80-Ly6C+）和中性粒细胞（Ly6G+）浸润到大脑，诱导脑膜炎发生。综上，本研究结果表明Clp是一个新的候选毒力蛋白，通过促进细菌的粘附，抑制细胞间紧密连接的表达，诱导脑微血管内皮细胞凋亡，从而破坏BBB增加其通透性。因此，SS2 Clp是细菌通过BBB所必需的，其结果为预防和治疗SS2诱导的脑膜炎补充了新理论依据。

Abstract:

Streptococcus suis serotype 2 (SS2) is an emerging zoonotic pathogen that causes meningitis in humans and pigs. It not only brings huge economic losses to the pig industry but also seriously threatens public health security. However, the mechanisms by which SS2 enters the brain and induces meningitis is not fully understood. Here, we investigated the role and mechanism of the SS2 collagenase-like protease (Clp) in promoting the passage of the bacterium across the blood-brain barrier (BBB). We found that SS2 Clp enhanced virulence and tissue colonization, and promoted the destruction of the BBB in mice. Compared with wild-type SS2, the ability of a Δclp mutant to cross human brain microvascular endothelial (hCMEC/D3) cell monolayers decreased, whereas the addition of recombinant protein rClp increased permeability. rClp also significantly promoted the adhesion of SS2 to hCMEC/D3, inhibited the expression of intercellular tight junction proteins ZO-1, Occludin, and Claudin-5 independent of its enzyme activity, and induced hCMEC/D3 apoptosis through the cell receptor ligand apoptosis and mitochondrial apoptosis pathways partly dependent on its enzyme activity, resulting in BBB destruction and increased permeability. Moreover, Clp increased macrophage (F4/80+), monocytes (F4/80-Ly6C+), and neutrophils (Ly6G+) infiltration into the brain after SS2 infection. Thus, SS2 Clp is required for the passage of the bacterium across the BBB, and the results, provide a theoretical basis for better prevention and treatment of SS2-induced meningitis.

Key words: Streptococcus suis serotype 2 , collagenase-like protease , Meningitis , blood brain barrier , permeability

Jikun Mei, Xuan Jiang, Fengyang Li, Zengshuai Wu, Tong Wu, Junhui Zhu, Hexiang Jiang, Ziheng Li, Na Li, Liancheng Lei. 猪链球菌2型胶原酶样蛋白酶增强血脑屏障通透性促进脑膜炎发生[J]. Journal of Integrative Agriculture, 2025, 24(11): 4379-4394.

Jikun Mei, Xuan Jiang, Fengyang Li, Zengshuai Wu, Tong Wu, Junhui Zhu, Hexiang Jiang, Ziheng Li, Na Li, Liancheng Lei. Streptococcus suis serotype 2 collagenase-like protease promotes meningitis by increasing blood–brain barrier permeability[J]. Journal of Integrative Agriculture, 2025, 24(11): 4379-4394.

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猪链球菌2型胶原酶样蛋白酶增强血脑屏障通透性促进脑膜炎发生

Streptococcus suis serotype 2 collagenase-like protease promotes meningitis by increasing blood–brain barrier permeability

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