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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| Published in | Emerging microbes & infections Vol. 8; no. 1; pp. 724 - 733 |
|---|---|
| Main Authors | , , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
Taylor & Francis
01.01.2019
Taylor & Francis Ltd Taylor & Francis Group |
| Subjects | |
| Online Access | Get full text |
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| Summary: | 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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| Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 These authors contributed equally. Y.L.Z. and Y.G. contributed equally to this work. Y.L.Z., Y.G., J.H.Z. and J.L.C. established the cell infection system. Y.L.Z., S.H.O., and C.R.C. collected the serum samples. Y.G., J.H.Z., J.L.C. and Y.L.Z. completed the cell-based assays for the serum samples. Y.L.Z., C.L.Y. and H.L.X. completed cell line evaluations. Y.L.Z. Y.B.W., S.H.O. performed the immunoassays. Y.L.Z., Y.G., S.H.O. and Q.Y. designed the experiments and performed data analysis. Y.L.Z., Y.G. and Q.Y. wrote the manuscript. T.Y.Z., T.C., C.R.C. and N.S.X. performed critical revision of the manuscript for important intellectual content. Q.Y., S.H.O, C.R.C. and N.S.X. supervised this project. Supplemental data for this article can be accessed at https://doi.org/10.1080/22221751.2019.1619485. |
| ISSN: | 2222-1751 2222-1751 |
| DOI: | 10.1080/22221751.2019.1619485 |