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

Anti-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...

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Autores principales: Zhang, Ya-Li, Gao, Ying, Cao, Jia-Li, Zhao, Jing-Hua, Zhang, Tian-Ying, Yang, Chuan-Lai, Xiong, Hua-Long, Wang, Ying-Bin, Ou, Shan-Hai, Cheng, Tong, Chen, Chang-Rong, Yuan, Quan, Xia, Ning-Shao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6542156/
https://www.ncbi.nlm.nih.gov/pubmed/31130075
http://dx.doi.org/10.1080/22221751.2019.1619485
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author Zhang, Ya-Li
Gao, Ying
Cao, Jia-Li
Zhao, Jing-Hua
Zhang, Tian-Ying
Yang, Chuan-Lai
Xiong, Hua-Long
Wang, Ying-Bin
Ou, Shan-Hai
Cheng, Tong
Chen, Chang-Rong
Yuan, Quan
Xia, Ning-Shao
author_facet Zhang, Ya-Li
Gao, Ying
Cao, Jia-Li
Zhao, Jing-Hua
Zhang, Tian-Ying
Yang, Chuan-Lai
Xiong, Hua-Long
Wang, Ying-Bin
Ou, Shan-Hai
Cheng, Tong
Chen, Chang-Rong
Yuan, Quan
Xia, Ning-Shao
author_sort Zhang, Ya-Li
collection PubMed
description Anti-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 (MDF(50)) 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 (R(2) = 0.473, p < 0.001). ROC analysis indicated that the optimal cutoff value of the qAnti-HBs to discriminate significant NAT (MDF(50) ≥ 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 (R(2) = 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; MDF(50): the maximum fold dilution required to produce ≥50% inhibition; IC50: half maximal inhibitory concentration
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spelling pubmed-65421562019-06-12 Robust in vitro assay for analyzing the neutralization activity of serum specimens against hepatitis B virus Zhang, Ya-Li Gao, Ying Cao, Jia-Li Zhao, Jing-Hua Zhang, Tian-Ying Yang, Chuan-Lai Xiong, Hua-Long Wang, Ying-Bin Ou, Shan-Hai Cheng, Tong Chen, Chang-Rong Yuan, Quan Xia, Ning-Shao Emerg Microbes Infect Article Anti-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 (MDF(50)) 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 (R(2) = 0.473, p < 0.001). ROC analysis indicated that the optimal cutoff value of the qAnti-HBs to discriminate significant NAT (MDF(50) ≥ 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 (R(2) = 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; MDF(50): the maximum fold dilution required to produce ≥50% inhibition; IC50: half maximal inhibitory concentration Taylor & Francis 2019-05-25 /pmc/articles/PMC6542156/ /pubmed/31130075 http://dx.doi.org/10.1080/22221751.2019.1619485 Text en © 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of Shanghai Shangyixun Cultural Communication Co., Ltd http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Article
Zhang, Ya-Li
Gao, Ying
Cao, Jia-Li
Zhao, Jing-Hua
Zhang, Tian-Ying
Yang, Chuan-Lai
Xiong, Hua-Long
Wang, Ying-Bin
Ou, Shan-Hai
Cheng, Tong
Chen, Chang-Rong
Yuan, Quan
Xia, Ning-Shao
Robust in vitro assay for analyzing the neutralization activity of serum specimens against hepatitis B virus
title Robust in vitro assay for analyzing the neutralization activity of serum specimens against hepatitis B virus
title_full Robust in vitro assay for analyzing the neutralization activity of serum specimens against hepatitis B virus
title_fullStr Robust in vitro assay for analyzing the neutralization activity of serum specimens against hepatitis B virus
title_full_unstemmed Robust in vitro assay for analyzing the neutralization activity of serum specimens against hepatitis B virus
title_short Robust in vitro assay for analyzing the neutralization activity of serum specimens against hepatitis B virus
title_sort robust in vitro assay for analyzing the neutralization activity of serum specimens against hepatitis b virus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6542156/
https://www.ncbi.nlm.nih.gov/pubmed/31130075
http://dx.doi.org/10.1080/22221751.2019.1619485
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