Analysis of epitopes on dengue virus envelope protein recognized by monoclonal antibodies and polyclonal human sera by a high throughput assay.
BACKGROUND: The envelope (E) protein of dengue virus (DENV) is the major target of neutralizing antibodies and vaccine development. While previous studies on domain III or domain I/II alone have reported several epitopes of monoclonal antibodies (mAbs) against DENV E protein, the possibility of inte...
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ftdoajarticles:oai:doaj.org/article:7dbe71a04b3f4f89b13f226148326584 2023-05-15T15:16:00+02:00 Analysis of epitopes on dengue virus envelope protein recognized by monoclonal antibodies and polyclonal human sera by a high throughput assay. Hong-En Lin Wen-Yang Tsai I-Ju Liu Pi-Chun Li Mei-Ying Liao Jih-Jin Tsai Yi-Chieh Wu Chih-Yun Lai Chih-Hsuan Lu Jyh-Hsiung Huang Gwong-Jen Chang Han-Chung Wu Wei-Kung Wang 2012-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0001447 https://doaj.org/article/7dbe71a04b3f4f89b13f226148326584 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC3250511?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0001447 https://doaj.org/article/7dbe71a04b3f4f89b13f226148326584 PLoS Neglected Tropical Diseases, Vol 6, Iss 1, p e1447 (2012) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2012 ftdoajarticles https://doi.org/10.1371/journal.pntd.0001447 2022-12-31T07:25:08Z BACKGROUND: The envelope (E) protein of dengue virus (DENV) is the major target of neutralizing antibodies and vaccine development. While previous studies on domain III or domain I/II alone have reported several epitopes of monoclonal antibodies (mAbs) against DENV E protein, the possibility of interdomain epitopes and the relationship between epitopes and neutralizing potency remain largely unexplored. METHODOLOGY/PRINCIPAL FINDINGS: We developed a dot blot assay by using 67 alanine mutants of predicted surface-exposed E residues as a systematic approach to identify epitopes recognized by mAbs and polyclonal sera, and confirmed our findings using a capture-ELISA assay. Of the 12 mouse mAbs tested, three recognized a novel epitope involving residues (Q211, D215, P217) at the central interface of domain II, and three recognized residues at both domain III and the lateral ridge of domain II, suggesting a more frequent presence of interdomain epitopes than previously appreciated. Compared with mAbs generated by traditional protocols, the potent neutralizing mAbs generated by a new protocol recognized multiple residues in A strand or residues in C strand/CC' loop of DENV2 and DENV1, and multiple residues in BC loop and residues in DE loop, EF loop/F strand or G strand of DENV1. The predominant epitopes of anti-E antibodies in polyclonal sera were found to include both fusion loop and non-fusion residues in the same or adjacent monomer. CONCLUSIONS/SIGNIFICANCE: Our analyses have implications for epitope-specific diagnostics and epitope-based dengue vaccines. This high throughput method has tremendous application for mapping both intra and interdomain epitopes recognized by human mAbs and polyclonal sera, which would further our understanding of humoral immune responses to DENV at the epitope level. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 6 1 e1447 |
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Directory of Open Access Journals: DOAJ Articles |
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English |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Hong-En Lin Wen-Yang Tsai I-Ju Liu Pi-Chun Li Mei-Ying Liao Jih-Jin Tsai Yi-Chieh Wu Chih-Yun Lai Chih-Hsuan Lu Jyh-Hsiung Huang Gwong-Jen Chang Han-Chung Wu Wei-Kung Wang Analysis of epitopes on dengue virus envelope protein recognized by monoclonal antibodies and polyclonal human sera by a high throughput assay. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
BACKGROUND: The envelope (E) protein of dengue virus (DENV) is the major target of neutralizing antibodies and vaccine development. While previous studies on domain III or domain I/II alone have reported several epitopes of monoclonal antibodies (mAbs) against DENV E protein, the possibility of interdomain epitopes and the relationship between epitopes and neutralizing potency remain largely unexplored. METHODOLOGY/PRINCIPAL FINDINGS: We developed a dot blot assay by using 67 alanine mutants of predicted surface-exposed E residues as a systematic approach to identify epitopes recognized by mAbs and polyclonal sera, and confirmed our findings using a capture-ELISA assay. Of the 12 mouse mAbs tested, three recognized a novel epitope involving residues (Q211, D215, P217) at the central interface of domain II, and three recognized residues at both domain III and the lateral ridge of domain II, suggesting a more frequent presence of interdomain epitopes than previously appreciated. Compared with mAbs generated by traditional protocols, the potent neutralizing mAbs generated by a new protocol recognized multiple residues in A strand or residues in C strand/CC' loop of DENV2 and DENV1, and multiple residues in BC loop and residues in DE loop, EF loop/F strand or G strand of DENV1. The predominant epitopes of anti-E antibodies in polyclonal sera were found to include both fusion loop and non-fusion residues in the same or adjacent monomer. CONCLUSIONS/SIGNIFICANCE: Our analyses have implications for epitope-specific diagnostics and epitope-based dengue vaccines. This high throughput method has tremendous application for mapping both intra and interdomain epitopes recognized by human mAbs and polyclonal sera, which would further our understanding of humoral immune responses to DENV at the epitope level. |
format |
Article in Journal/Newspaper |
author |
Hong-En Lin Wen-Yang Tsai I-Ju Liu Pi-Chun Li Mei-Ying Liao Jih-Jin Tsai Yi-Chieh Wu Chih-Yun Lai Chih-Hsuan Lu Jyh-Hsiung Huang Gwong-Jen Chang Han-Chung Wu Wei-Kung Wang |
author_facet |
Hong-En Lin Wen-Yang Tsai I-Ju Liu Pi-Chun Li Mei-Ying Liao Jih-Jin Tsai Yi-Chieh Wu Chih-Yun Lai Chih-Hsuan Lu Jyh-Hsiung Huang Gwong-Jen Chang Han-Chung Wu Wei-Kung Wang |
author_sort |
Hong-En Lin |
title |
Analysis of epitopes on dengue virus envelope protein recognized by monoclonal antibodies and polyclonal human sera by a high throughput assay. |
title_short |
Analysis of epitopes on dengue virus envelope protein recognized by monoclonal antibodies and polyclonal human sera by a high throughput assay. |
title_full |
Analysis of epitopes on dengue virus envelope protein recognized by monoclonal antibodies and polyclonal human sera by a high throughput assay. |
title_fullStr |
Analysis of epitopes on dengue virus envelope protein recognized by monoclonal antibodies and polyclonal human sera by a high throughput assay. |
title_full_unstemmed |
Analysis of epitopes on dengue virus envelope protein recognized by monoclonal antibodies and polyclonal human sera by a high throughput assay. |
title_sort |
analysis of epitopes on dengue virus envelope protein recognized by monoclonal antibodies and polyclonal human sera by a high throughput assay. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2012 |
url |
https://doi.org/10.1371/journal.pntd.0001447 https://doaj.org/article/7dbe71a04b3f4f89b13f226148326584 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 6, Iss 1, p e1447 (2012) |
op_relation |
http://europepmc.org/articles/PMC3250511?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0001447 https://doaj.org/article/7dbe71a04b3f4f89b13f226148326584 |
op_doi |
https://doi.org/10.1371/journal.pntd.0001447 |
container_title |
PLoS Neglected Tropical Diseases |
container_volume |
6 |
container_issue |
1 |
container_start_page |
e1447 |
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1766346314643144704 |