A high-throughput neutralizing assay for antibodies and sera evaluation against Epstein-Barr virus

被引:2
作者
Zhong, Ling [1 ]
Krummenacher, Claude [4 ]
Zhang, Wanlin [1 ]
Hong, Junping [3 ]
Feng, Qisheng [1 ]
Zhao, Qinjian [2 ]
Chen, Yixin [3 ]
Zeng, Mu-Sheng [1 ]
Zeng, Yi-Xin [1 ]
Xu, Miao [1 ]
Zhang, Xiao [1 ,2 ]
机构
[1] Sun Yat Sen Univ, Canc Ctr, Collaborat Innovat Ctr Canc Med, Dept Expt Res,State Key Lab Oncol South China, Guangzhou, Guangdong, Peoples R China
[2] Chongqing Med Univ, Coll Pharm, Chongqing, Peoples R China
[3] Xiamen Univ, Sch Publ Hlth, Natl Inst Diagnost & Vaccine Dev Infect Dis, State Key Lab Mol Vaccinol & Mol Diagnost, Xiamen, Fujian, Peoples R China
[4] Rowan Univ, Dept Biol & Biomed Sci, Glassboro, NJ USA
基金
中国国家自然科学基金;
关键词
Epstein-Barr virus; High-throughput assay; Neutralizing antibodies; Human sera; Monkey sera; Anti-gHgL antibody CL59; INFECTIOUS-MONONUCLEOSIS; GP350; VACCINE; B-CELLS; EBV; RECEPTOR; TRIAL; RISK; IMMUNOGENICITY; ANTIGEN; FUSION;
D O I
10.1186/s12985-022-01911-1
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Background Epstein-Barr virus (EBV) is a wide-spread human herpesvirus that is highly associated with infectious mononucleosis and several malignancies. Evaluation of EBV neutralizing antibody titers is important for serological studies, vaccine development and monoclonal antibody screening. The traditional method based on antibody inhibition of EBV transformation of B cells is very time-consuming. A more practical flow cytometry-based (FCM) approach to evaluate neutralizing titers is not amenable to achieving high-throughput evaluation of large-scale samples. A high-throughput approach is urgently needed. Results Here, we present a rapid and high-throughput method based on high content imaging system (HCIS) analysis. EBV titers determined by the HCIS-based assay were similar to those obtained by the FCM-based assay. Neutralizing titers of sera and monoclonal antibodies measured by the HCIS-based assay strongly correlated with titers measured by the FCM-based assay. HCIS assays showed a strong correlation between B cell infection neutralizing titers and the anti-gp350 IgG titers in healthy EBV carriers and monkey sera. Finally, anti-gHgL IgG titers from sera of healthy EBV carriers significantly correlated with epithelial cell infection neutralizing titers. Conclusions This HCIS-based assay is a high-throughput assay to determine viral titers and evaluate neutralizing potentials of sera and monoclonal antibodies. This HCIS-based assay will aid the development of vaccines and therapeutic monoclonal antibody against EBV.
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页数:15
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