摘要
目的探讨脂蛋白特异性抗体通过血脑屏障及其在抑制金黄色葡萄球菌脑脓肿形成中的作用。方法利用大肠埃希菌表达并纯化重组金葡菌脂蛋白SA2275,制备特异性小鼠多克隆抗体(anti-SA2275)。制作金葡菌感染脑脓肿模型,利用ELISA法检测经腹腔注射的anti-SA2275在小鼠脑组织中的滴度,评估抗体通过血脑屏障的能力;再利用梯度稀释细菌计数法检测脑脓肿中的细菌载量,对脑脓肿进行连续病理切片检测脑脓肿的大小,评价抗体anti-SA2275治疗金葡菌脑脓肿的效果。结果成功获得高纯度SA2275重组蛋白;免疫小鼠获得anti-SA2275,效价为1:51200;anti-SA2275注射金葡菌脑脓肿模型小鼠后,在动物脑组织中检测到抗体的存在;治疗组小鼠脑脓肿大小和菌载量显著低于阴性对照组。结论在金葡菌感染小鼠所致的脑脓肿形成过程中,脂蛋白SA2275特异性抗体可以通过血脑屏障分布至脑组织,显著抑制脑脓肿的形成及细菌的定植,具有治疗金葡菌脑脓肿的应用前景。
Objective To investigate the ability of specific polyclonal antibodies against Staphylococcus aureus(S.aureus)lipoprotein SA2275 to penetrate the blood-brain barrier for inhibiting the formation of S.aureus-induced brain abscess in mice.Methods We immunized BALB/c mouse mice with purified recombinant S.aureus lipoprotein SA2275 expressed in Escherichia coli to obtain specific mouse polyclonal antibodies against SA2275(anti-SA2275).In a C57BL/6 mouse model of brain abscess induced by injecting S.aureus strain Newman into the brain,we tested the titer of anti-SA2275 in the brain tissue using ELISA following intraperitoneal injection of the antibodies to evaluate the ability of the antibodies to pass through the blood-brain barrier.The bacterial load in the mouse brain was determined by gradient dilution bacterial counting,and the size of the brain abscess was evaluated on serial sections.Results The recombinant SA2275 protein with a high purity was successfully obtained.The results of ELISA showed that immunization of the mice with the recombinant SA2275 protein resulted in a high titer of anti-SA2275 reaching 1:51200.In the mouse models of S.aureus-induced brain abscess,the presence of the antibodies was detected in the brain tissue after intraperitoneal injection of anti-SA2275.Treatment with anti-SA2275,as compared with blank control mouse serum,significantly reduced the size of the brain abscess and lowered the bacterial load in the brain tissues of the mice.Conclusion In mouse models of S.aureus-induced brain abscess,intraperitoneally injected lipoprotein SA2275-specific antibodies can penetrate the blood-brain barrier to reach the brain tissue and inhibit the formation of brain abscess and bacterial colonization,suggesting the potential of the antibodies for treatment of S.aureus-induced brain abscess.
作者
郑颖
杨裔
彭华刚
饶一凡
胡珍
胡启文
钟莲梅
饶贤才
尚伟龙
周人杰
ZHENG Ying;YANG Yi;PENG Huagang;RAO Yifan;HU Zhen;HU Qiwen;ZHONG Lianmei;RAO Xiancai;SHANG Weilong;ZHOU Renjie(Department of Neurology,First Affiliated Hospital of Kunming Medical University,Kunming,Yunnan Province,650032;Department of Microbiology,Chongqing Laboratory of Microbiological Engineering,College of Basic Medical Sciences,Army Medical University(Third Military Medical University),Chongqing,400038;School of Life Sciences,Institute of Modern Biopharmaceuticals,State Key Laboratory Breeding Base of Eco-Environment and Bio-Resource of the Three Gorges Area,Key Laboratory of Eco-environments in the Three Gorges Reservoir Region of Ministry of Education,Southwest University,Chongqing,400715;Department of Emergency Medicine,Second Affiliated Hospital,Army Medical University(Third Military Medical University),400037,China)
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2020年第16期1655-1662,共8页
Journal of Third Military Medical University
基金
国家自然科学基金青年科学基金(81701967)。
关键词
金黄色葡萄球菌
脑脓肿
抗体
血脑屏障
Staphylococcus aureus
antibodies
brain abscess
blood-brain barrier
