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Patient-derived monoclonal antibody neutralizes HCV infection in vitro and vivo without generating escape mutants
In recent years, new direct-acting antivirals for hepatitis C virus (HCV) have been approved, but hepatitis C continues to pose a threat to human health. It is important to develop neutralizing anti-HCV antibodies to prevent medical and accidental infection, such as might occur via liver transplanta...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9499215/ https://www.ncbi.nlm.nih.gov/pubmed/36137152 http://dx.doi.org/10.1371/journal.pone.0274283 |
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author | Yokokawa, Hiroshi Shinohara, Midori Teraoka, Yuji Imamura, Michio Nakamura, Noriko Watanabe, Noriyuki Date, Tomoko Aizaki, Hideki Iwamura, Tomokatsu Narumi, Hideki Chayama, Kazuaki Wakita, Takaji |
author_facet | Yokokawa, Hiroshi Shinohara, Midori Teraoka, Yuji Imamura, Michio Nakamura, Noriko Watanabe, Noriyuki Date, Tomoko Aizaki, Hideki Iwamura, Tomokatsu Narumi, Hideki Chayama, Kazuaki Wakita, Takaji |
author_sort | Yokokawa, Hiroshi |
collection | PubMed |
description | In recent years, new direct-acting antivirals for hepatitis C virus (HCV) have been approved, but hepatitis C continues to pose a threat to human health. It is important to develop neutralizing anti-HCV antibodies to prevent medical and accidental infection, such as might occur via liver transplantation of chronic HCV patients and needle-stick accidents in the clinic. In this study, we sought to obtain anti-HCV antibodies using phage display screening. Phages displaying human hepatocellular carcinoma patient-derived antibodies were screened by 4 rounds of biopanning with genotype-1b and -2a HCV envelope E2 protein adsorbed to magnetic beads. The three antibodies obtained from this screen had reactivity against E2 proteins derived from both genotype-1b and -2a strains. However, in epitope analysis, these antibodies did not recognize linear peptides from an overlapping E2 epitope peptide library, and did not bind to denatured E2 protein. In addition, these antibodies showed cross-genotypic neutralizing activity against genotype-1a, -1b, -2a, and -3a cell culture-generated infectious HCV particles (HCVcc). Moreover, emergence of viral escape mutants was not observed after repeated rounds of passaging of HCV-infected cells in the presence of one such antibody, e2d066. Furthermore, injection of the e2d066 antibody into human hepatocyte-transplanted immunodeficient mice inhibited infection by J6/JFH-1 HCVcc. In conclusion, we identified conformational epitope-recognizing, cross-genotypic neutralizing antibodies using phage display screening. Notably, e2d066 antibody did not select for escape mutant emergence in vitro and demonstrated neutralizing activity in vivo. Our results suggested that these antibodies may serve as prophylactic and therapeutic agents. |
format | Online Article Text |
id | pubmed-9499215 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-94992152022-09-23 Patient-derived monoclonal antibody neutralizes HCV infection in vitro and vivo without generating escape mutants Yokokawa, Hiroshi Shinohara, Midori Teraoka, Yuji Imamura, Michio Nakamura, Noriko Watanabe, Noriyuki Date, Tomoko Aizaki, Hideki Iwamura, Tomokatsu Narumi, Hideki Chayama, Kazuaki Wakita, Takaji PLoS One Research Article In recent years, new direct-acting antivirals for hepatitis C virus (HCV) have been approved, but hepatitis C continues to pose a threat to human health. It is important to develop neutralizing anti-HCV antibodies to prevent medical and accidental infection, such as might occur via liver transplantation of chronic HCV patients and needle-stick accidents in the clinic. In this study, we sought to obtain anti-HCV antibodies using phage display screening. Phages displaying human hepatocellular carcinoma patient-derived antibodies were screened by 4 rounds of biopanning with genotype-1b and -2a HCV envelope E2 protein adsorbed to magnetic beads. The three antibodies obtained from this screen had reactivity against E2 proteins derived from both genotype-1b and -2a strains. However, in epitope analysis, these antibodies did not recognize linear peptides from an overlapping E2 epitope peptide library, and did not bind to denatured E2 protein. In addition, these antibodies showed cross-genotypic neutralizing activity against genotype-1a, -1b, -2a, and -3a cell culture-generated infectious HCV particles (HCVcc). Moreover, emergence of viral escape mutants was not observed after repeated rounds of passaging of HCV-infected cells in the presence of one such antibody, e2d066. Furthermore, injection of the e2d066 antibody into human hepatocyte-transplanted immunodeficient mice inhibited infection by J6/JFH-1 HCVcc. In conclusion, we identified conformational epitope-recognizing, cross-genotypic neutralizing antibodies using phage display screening. Notably, e2d066 antibody did not select for escape mutant emergence in vitro and demonstrated neutralizing activity in vivo. Our results suggested that these antibodies may serve as prophylactic and therapeutic agents. Public Library of Science 2022-09-22 /pmc/articles/PMC9499215/ /pubmed/36137152 http://dx.doi.org/10.1371/journal.pone.0274283 Text en © 2022 Yokokawa et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Yokokawa, Hiroshi Shinohara, Midori Teraoka, Yuji Imamura, Michio Nakamura, Noriko Watanabe, Noriyuki Date, Tomoko Aizaki, Hideki Iwamura, Tomokatsu Narumi, Hideki Chayama, Kazuaki Wakita, Takaji Patient-derived monoclonal antibody neutralizes HCV infection in vitro and vivo without generating escape mutants |
title | Patient-derived monoclonal antibody neutralizes HCV infection in vitro and vivo without generating escape mutants |
title_full | Patient-derived monoclonal antibody neutralizes HCV infection in vitro and vivo without generating escape mutants |
title_fullStr | Patient-derived monoclonal antibody neutralizes HCV infection in vitro and vivo without generating escape mutants |
title_full_unstemmed | Patient-derived monoclonal antibody neutralizes HCV infection in vitro and vivo without generating escape mutants |
title_short | Patient-derived monoclonal antibody neutralizes HCV infection in vitro and vivo without generating escape mutants |
title_sort | patient-derived monoclonal antibody neutralizes hcv infection in vitro and vivo without generating escape mutants |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9499215/ https://www.ncbi.nlm.nih.gov/pubmed/36137152 http://dx.doi.org/10.1371/journal.pone.0274283 |
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