Cargando…
Antibody epitope profiling of the KSHV LANA protein using VirScan
The humoral antibody response against Kaposi sarcoma-associated herpesvirus (KSHV) in infected individuals has been characterized demonstrating the latency-associated nuclear antigen (LANA) as the most antigenic KSHV protein. Despite the antigenicity of the protein, specific LANA epitopes have not b...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9810164/ https://www.ncbi.nlm.nih.gov/pubmed/36534707 http://dx.doi.org/10.1371/journal.ppat.1011033 |
_version_ | 1784863252875837440 |
---|---|
author | Bennett, Sydney J. Yalcin, Dicle Privatt, Sara R. Ngalamika, Owen Lidenge, Salum J. West, John T. Wood, Charles |
author_facet | Bennett, Sydney J. Yalcin, Dicle Privatt, Sara R. Ngalamika, Owen Lidenge, Salum J. West, John T. Wood, Charles |
author_sort | Bennett, Sydney J. |
collection | PubMed |
description | The humoral antibody response against Kaposi sarcoma-associated herpesvirus (KSHV) in infected individuals has been characterized demonstrating the latency-associated nuclear antigen (LANA) as the most antigenic KSHV protein. Despite the antigenicity of the protein, specific LANA epitopes have not been systematically characterized. Here, we utilized a bacteriophage T7 library, which displays 56-amino acid KSHV LANA peptides with 28-amino acid overlap (VirScan), to define those epitopes in LANA targeted by antibodies from a cohort of 62 sub-Saharan African Kaposi sarcoma (KS) patients and 22 KSHV-infected asymptomatic controls. Intra- and inter-patient breadth and magnitude of the anti-LANA responses were quantified at the peptide and amino acid levels. From these data, we derived a detailed epitope annotation of the entire LANA protein, with a high-resolution focus on the N- and C-termini. Overall, the central repeat region was highly antigenic, but the responses to this region could not be confidently mapped due to its high variability. The highly conserved N-terminus was targeted with low breadth and magnitude. In a minority of individuals, antibodies specific to the nuclear localization sequence and a portion of the proline-rich regions of the N-terminus were evident. In contrast, the first half of the conserved C-terminal domain was consistently targeted with high magnitude. Unfortunately, this region was not included in LANA partial C-terminal crystal structures, however, it was predicted to adopt predominantly random-coil structure. Coupled with functional and secondary structure domain predictions, VirScan revealed fine resolution epitope mapping of the N- and C-terminal domains of LANA that is consistent with previous antigenicity studies and may prove useful to correlate KSHV humoral immunity with pathogenesis. |
format | Online Article Text |
id | pubmed-9810164 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-98101642023-01-04 Antibody epitope profiling of the KSHV LANA protein using VirScan Bennett, Sydney J. Yalcin, Dicle Privatt, Sara R. Ngalamika, Owen Lidenge, Salum J. West, John T. Wood, Charles PLoS Pathog Research Article The humoral antibody response against Kaposi sarcoma-associated herpesvirus (KSHV) in infected individuals has been characterized demonstrating the latency-associated nuclear antigen (LANA) as the most antigenic KSHV protein. Despite the antigenicity of the protein, specific LANA epitopes have not been systematically characterized. Here, we utilized a bacteriophage T7 library, which displays 56-amino acid KSHV LANA peptides with 28-amino acid overlap (VirScan), to define those epitopes in LANA targeted by antibodies from a cohort of 62 sub-Saharan African Kaposi sarcoma (KS) patients and 22 KSHV-infected asymptomatic controls. Intra- and inter-patient breadth and magnitude of the anti-LANA responses were quantified at the peptide and amino acid levels. From these data, we derived a detailed epitope annotation of the entire LANA protein, with a high-resolution focus on the N- and C-termini. Overall, the central repeat region was highly antigenic, but the responses to this region could not be confidently mapped due to its high variability. The highly conserved N-terminus was targeted with low breadth and magnitude. In a minority of individuals, antibodies specific to the nuclear localization sequence and a portion of the proline-rich regions of the N-terminus were evident. In contrast, the first half of the conserved C-terminal domain was consistently targeted with high magnitude. Unfortunately, this region was not included in LANA partial C-terminal crystal structures, however, it was predicted to adopt predominantly random-coil structure. Coupled with functional and secondary structure domain predictions, VirScan revealed fine resolution epitope mapping of the N- and C-terminal domains of LANA that is consistent with previous antigenicity studies and may prove useful to correlate KSHV humoral immunity with pathogenesis. Public Library of Science 2022-12-19 /pmc/articles/PMC9810164/ /pubmed/36534707 http://dx.doi.org/10.1371/journal.ppat.1011033 Text en © 2022 Bennett 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 Bennett, Sydney J. Yalcin, Dicle Privatt, Sara R. Ngalamika, Owen Lidenge, Salum J. West, John T. Wood, Charles Antibody epitope profiling of the KSHV LANA protein using VirScan |
title | Antibody epitope profiling of the KSHV LANA protein using VirScan |
title_full | Antibody epitope profiling of the KSHV LANA protein using VirScan |
title_fullStr | Antibody epitope profiling of the KSHV LANA protein using VirScan |
title_full_unstemmed | Antibody epitope profiling of the KSHV LANA protein using VirScan |
title_short | Antibody epitope profiling of the KSHV LANA protein using VirScan |
title_sort | antibody epitope profiling of the kshv lana protein using virscan |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9810164/ https://www.ncbi.nlm.nih.gov/pubmed/36534707 http://dx.doi.org/10.1371/journal.ppat.1011033 |
work_keys_str_mv | AT bennettsydneyj antibodyepitopeprofilingofthekshvlanaproteinusingvirscan AT yalcindicle antibodyepitopeprofilingofthekshvlanaproteinusingvirscan AT privattsarar antibodyepitopeprofilingofthekshvlanaproteinusingvirscan AT ngalamikaowen antibodyepitopeprofilingofthekshvlanaproteinusingvirscan AT lidengesalumj antibodyepitopeprofilingofthekshvlanaproteinusingvirscan AT westjohnt antibodyepitopeprofilingofthekshvlanaproteinusingvirscan AT woodcharles antibodyepitopeprofilingofthekshvlanaproteinusingvirscan |