Robust in vitro assay for analyzing the neutralization activity of serum specimens against hepatitis B virus

Ya-Li Zhang; Ying Gao; Jia-Li Cao; Jing-Hua Zhao; Tian-Ying Zhang; Chuan-Lai Yang; Hua-Long Xiong; Ying-Bin Wang; Shan-Hai Ou; Tong Cheng; Chang-Rong Chen; Quan Yuan; Ning-Shao Xia

doi:10.1080/22221751.2019.1619485

Emerging Microbes and Infections (Jan 2019)

Robust in vitro assay for analyzing the neutralization activity of serum specimens against hepatitis B virus

Ya-Li Zhang,
Ying Gao,
Jia-Li Cao,
Jing-Hua Zhao,
Tian-Ying Zhang,
Chuan-Lai Yang,
Hua-Long Xiong,
Ying-Bin Wang,
Shan-Hai Ou,
Tong Cheng,
Chang-Rong Chen,
Quan Yuan,
Ning-Shao Xia

Affiliations

Ya-Li Zhang: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, People’s Republic of China
Ying Gao: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, People’s Republic of China
Jia-Li Cao: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, People’s Republic of China
Jing-Hua Zhao: School of Life Science, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiamen, People’s Republic of China
Tian-Ying Zhang: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, People’s Republic of China
Chuan-Lai Yang: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, People’s Republic of China
Hua-Long Xiong: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, People’s Republic of China
Ying-Bin Wang: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, People’s Republic of China
Shan-Hai Ou: Xiamen Blood Service, Xiamen, People’s Republic of China
Tong Cheng: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, People’s Republic of China
Chang-Rong Chen: Xiamen Haicang Hospital, Xiamen, People’s Republic of China
Quan Yuan: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, People’s Republic of China
Ning-Shao Xia: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, People’s Republic of China

DOI: https://doi.org/10.1080/22221751.2019.1619485
Journal volume & issue: Vol. 8, no. 1
pp. 724 – 733

Abstract

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ABSTRACTAnti-HBs is a well-known marker of protective capability against HBV. However, little is known about the association between the qAnti-HBs determined by immunoassays and the neutralization activity (NAT) derived from functional assays. We developed an in vitro assay for direct measurement of the NAT of human sera. The new assay was highly sensitive, with an analytical sensitivity of 9.6 ± 1.3 mIU/mL for the HBIG standard. For serum detection, the maximum fold dilution required to produce ≥50% inhibition (MDF50) of HBV infection was used as the quantitative index. In vitro NAT evaluations were conducted for a cohort of 164 HBV-free healthy individuals. The results demonstrated that the NAT positively correlated with the qAnti-HBs (R2 = 0.473, p < 0.001). ROC analysis indicated that the optimal cutoff value of the qAnti-HBs to discriminate significant NAT (MDF50 ≥ 8) was 62.9 mIU/mL, with an AUROC of 0.920. Additionally, we found that the qAnti-HBc was another independent parameter positively associated with the NAT (R2 = 0.300, p < 0.001), which suggested that antibodies against other HBV proteins generated by previous HBV exposure possibly also contribute to the NAT. In summary, the new cell-based assay provides a robust tool to analyse the anti-HBV NAT.Abbreviations: HBV: Hepatitis B virus; HBsAg: Hepatitis B surface antigen; Anti-HBs: Hepatitis B surface antibody; HBeAg: Hepatitis B e antigen; Anti-HBc: Hepatitis B core antibody; qAnti-HBs: quantitative hepatitis B surface antibody; qAnti-HBc: quantitative hepatitis B core antibody; qHBeAg: quantitative hepatitis B e antigen; NAT: neutralization activity; HBIG: hepatitis B immune globulin; NTCP: Na+-taurocholate cotransporting polypeptide; IRES: internal ribosome entry site; ccHBV: cell culture derived hepatitis B virus; GE/cell: genome equivalent per cell; MOI: multiplicity of infection; Dpi: day post infection; HepG2-TetOn: a HepG2-derived cell line that expresses the doxycycline-regulated transactivator; ROC: receiver operating characteristic curve; AUROC: area under receiver operating characteristic curve; LLOQ: the lower limits of quantification; MDF50: the maximum fold dilution required to produce ≥50% inhibition; IC50: half maximal inhibitory concentration

Published in Emerging Microbes and Infections

ISSN: 2222-1751 (Online)
Publisher: Taylor & Francis Group
Country of publisher: United Kingdom
LCC subjects: Medicine: Internal medicine: Infectious and parasitic diseases; Science: Microbiology
Website: https://www.tandfonline.com/journals/temi

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