Development and characterization of neutralizing monoclonal antibodies against clinically relevant human adenovirus subgroup B serotypes

Yuelin Wang; Wei Dou; Yingying Chen; Ke Liu; Mengjun Li; Tyuji Hoshino; Yushan Jiang; Vladislav Victorovich Khrustalev; Alexey V. Churov; Alexander N. Orekhov; Wei Zhao; Ling Li; Yang Yang; Chenguang Shen

doi:10.1016/j.virs.2026.03.017

April 2026

. doi: 10.1016/j.virs.2026.03.017

Citation: Yuelin Wang, Wei Dou, Yingying Chen, Ke Liu, Mengjun Li, Tyuji Hoshino, Yushan Jiang, Vladislav Victorovich Khrustalev, Alexey V. Churov, Alexander N. Orekhov, Wei Zhao, Ling Li, Yang Yang, Chenguang Shen. Development and characterization of neutralizing monoclonal antibodies against clinically relevant human adenovirus subgroup B serotypes .VIROLOGICA SINICA, 2026, 41(2) : 404-412. http://dx.doi.org/10.1016/j.virs.2026.03.017

临床相关B亚属人腺病毒中和单克隆抗体的制备与鉴定

王跃霖 ^a,b ,
窦洧 ^a ,
陈莹莹 ^a,c ,
刘可 ^a ,
李梦君 ^a ,
Tyuji Hoshino ^d ,
蒋宇珊 ^a ,
Vladislav Victorovich Khrustalev ^e ,
Alexey V Churov ^f ,
Alexander N Orekhov ^f ,
赵卫 ^a,, ,
李凌 ^a,c,, ,
杨扬 ^b,, ,
沈晨光 ^a,g,h,,

a 南方医科大学公共卫生学院, 广东省热带病研究重点实验室, 生物安全三级实验室(广东);
b 南方科技大学第二附属医院(深圳市第三人民医院), 国家感染性疾病临床医学研究中心;
c 南方医科大学基础医学院;
d 千叶大学药学研究院, 日本;
e 白俄罗斯国立医科大学普通化学系, 白俄罗斯;
f 病理学与病理生理学研究所, 血管病理学实验室, 俄罗斯;
g南方医科大学珠江医院检验医学科;
h 南方医科大学, 华南传染病研究教育部重点实验室

通讯作者： 赵卫, zhaowei@smu.edu.cn; 李凌, liling@smu.edu.cn; 杨扬, young@mail.sustech.edu.cn; 沈晨光, a124965468@smu.edu.cn

摘要

人腺病毒（HAdVs）特别是B亚属3型和55型人腺病毒，是引起严重呼吸道疾病的重要病原，但目前临床上尚缺乏特异性治疗药物。尽管中和单克隆抗体（nMAbs）具有良好的治疗前景，但针对上述型别的高效中和抗体仍未被充分表征。本研究旨在制备针对HAdV-3和HAdV-55的特异性nMAbs并评价其有效性。通过使用HAdV-3、HAdV-55病毒颗粒或重组纤毛蛋白球行结构域（fiber knob）免疫小鼠，研究团队经系统筛选获得了11株特异性单抗，其中6株针对HAdV-3，5株针对HAdV-55/-7纤毛结节。体外实验显示，4株nMAbs具有强效的中和活性：针对HAdV-3的13F12、8D2和3A3（IC50分别为3.8、15.1和14.9 μg/mL），以及针对HAdV-55的8F2（IC50为30.4 μg/mL）。蛋白印迹分析证实，13F12和8F2靶向纤毛蛋白（fiber），而8D2和3A3则结合六邻体蛋白（hexon）。此外，树鼩体内评估表明，13F12能显著降低HAdV-3感染树鼩鼻甲中的病毒载量，并有效减轻肺部病理损伤。在机制上，所有中和单克隆抗体通过阻断病毒附着抑制感染（P < 0.01）。综上所述，本研究凸显了以病毒入侵过程为靶点的治疗价值，并证实单抗13F12是预防和治疗HAdV感染极具潜力的候选抗体。
- 人腺病毒（HAdV）
- , 中和抗体
- , 单克隆抗体
- , 抗病毒治疗
- , 呼吸道感染
- , B亚属血清型

Development and characterization of neutralizing monoclonal antibodies against clinically relevant human adenovirus subgroup B serotypes

a. BSL-3 Laboratory (Guangdong), Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou 510515, China;
b. National Clinical Research Center for Infectious Disease, Shenzhen Third People's Hospital, Second Hospital Affiliated to Southern University of Science and Technology, Shenzhen 518112, China;
c. School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China;
d. Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675, Japan;
e. Department of General Chemistry, Belarusian State Medical University, Minsk 220045, Belarus;
f. Laboratory of Angiopathology, Institute of General Pathology and Pathophysiology, Moscow 125315, Russia;
g. Department of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, China;
h. Key Laboratory of Infectious Diseases Research in South China (Southern Medical University), Ministry of Education, Guangzhou 510515, China

Corresponding author: Wei Zhao, zhaowei@smu.edu.cn Ling Li, liling@smu.edu.cn Yang Yang, young@mail.sustech.edu.cn Chenguang Shen, a124965468@smu.edu.cn

Abstract

Human adenoviruses (HAdVs), particularly subgroup B serotypes HAdV-3 and HAdV-55, are associated with severe respiratory disease and currently lack targeted therapies. While neutralizing monoclonal antibodies (nMAbs) offer promising therapeutic potential, the specific nMAbs targeting these serotypes remain poorly characterized. Therefore, this study aimed to generate and evaluate the efficacy of serotype-specific nMAbs against HAdV-3 and HAdV-55. Mice were immunized with HAdV-3 virions, HAdV-55 virions, or recombinant fiber knob proteins (HAdV-55/-7) for the generation of serotype-specific nMAbs, and their efficacy was systematically evaluated using in vitro assays and an in vivo tree shrew model. Through comprehensive screening, eleven MAbs were identified with specificity against HAdV-3 (six clones) or HAdV-55/-7 fiber knob (five clones). Four nMAbs exhibited potent neutralizing activity: 13F12 (half-maximal inhibitory concentration, IC₅₀: 3.8 μg/mL), 8D2 (IC₅₀: 15.1 μg/mL), 3A3 (IC₅₀: 14.9 μg/mL) against HAdV-3 virions, and 8F2 with neutralizing efficacy against HAdV-55 virions (IC₅₀: 30.4 μg/mL). Western blot analysis revealed that MAbs 13F12 and 8F2 targeted the fiber protein, whereas 8D2 and 3A3 bound to the hexon protein. Furthermore, in vivo evaluations demonstrated that 13F12 significantly reduced viral loads in nasal turbinates and attenuated lung pathology in HAdV-3-infected tree shrews. Mechanistically, all tested anti-HAdV-3 nMAbs inhibited infection by blocking viral attachment (P < 0.01 vs. controls). In conclusion, this study underscores the therapeutic potential of targeting viral entry and highlights 13F12 as a promising candidate for HAdV prophylaxis.
- Human adenovirus (HAdV)
- , Neutralizing antibody
- , Monoclonal antibody
- , Antiviral therapy
- , Respiratory infection
- , B subgroup serotypes

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10.1016j.virs.2026.03.017-ESM.docx