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Linear and conformational determinants of visceral leishmaniasis diagnostic antigens rK28 and rK39
BACKGROUND: Recombinant antigens rK39 (based on kinesin sequence) and rK28 (comprising kinesin and HASPB sequences) are a mainstay of serological diagnosis for visceral leishmaniasis (VL). However, their key epitopes and the significance of their structural conformation are not clearly defined, part...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9587664/ https://www.ncbi.nlm.nih.gov/pubmed/36273150 http://dx.doi.org/10.1186/s13071-022-05495-1 |
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author | Simonson, Poppy Bhattacharyya, Tapan El-Safi, Sayda Miles, Michael A. |
author_facet | Simonson, Poppy Bhattacharyya, Tapan El-Safi, Sayda Miles, Michael A. |
author_sort | Simonson, Poppy |
collection | PubMed |
description | BACKGROUND: Recombinant antigens rK39 (based on kinesin sequence) and rK28 (comprising kinesin and HASPB sequences) are a mainstay of serological diagnosis for visceral leishmaniasis (VL). However, their key epitopes and the significance of their structural conformation are not clearly defined, particularly in relation to reported cross-reactivity with sera from patients with malaria, schistosomiasis, and tuberculosis. METHODS: To assess the effect of conformation on antigenicity with Sudanese VL sera, antigens rK39 and rK28 were heat-denatured at 95 °C for 10 min and then assayed by enzyme-linked immunosorbent assay (ELISA). Amino acid sequences of rK39 and rK28 were submitted to NCBI BLASTp to assess homology with Plasmodium, Schistosoma, and Mycobacterium. RESULTS: Heat denaturation significantly diminished the antigenicity of rK39 compared to non-denatured antigen (P = 0.001), but not for rK28 (P = 0.275). In BLASTp searches, HASPB sequences from rK28 had similarities with sequences from Plasmodium, encompassing software-predicted B-cell epitopes. CONCLUSIONS: The antigenicity of rK39 appears to be dependent on structural conformation, whereas that of rK28 depends on linear sequence. HASPB sequence homology with Plasmodium may be responsible for the reported cross-reactivity of rK28 with malaria sera. Further work is warranted to refine the specificity of these antigens. GRAPHICAL ABSTRACT: [Image: see text] |
format | Online Article Text |
id | pubmed-9587664 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-95876642022-10-23 Linear and conformational determinants of visceral leishmaniasis diagnostic antigens rK28 and rK39 Simonson, Poppy Bhattacharyya, Tapan El-Safi, Sayda Miles, Michael A. Parasit Vectors Research BACKGROUND: Recombinant antigens rK39 (based on kinesin sequence) and rK28 (comprising kinesin and HASPB sequences) are a mainstay of serological diagnosis for visceral leishmaniasis (VL). However, their key epitopes and the significance of their structural conformation are not clearly defined, particularly in relation to reported cross-reactivity with sera from patients with malaria, schistosomiasis, and tuberculosis. METHODS: To assess the effect of conformation on antigenicity with Sudanese VL sera, antigens rK39 and rK28 were heat-denatured at 95 °C for 10 min and then assayed by enzyme-linked immunosorbent assay (ELISA). Amino acid sequences of rK39 and rK28 were submitted to NCBI BLASTp to assess homology with Plasmodium, Schistosoma, and Mycobacterium. RESULTS: Heat denaturation significantly diminished the antigenicity of rK39 compared to non-denatured antigen (P = 0.001), but not for rK28 (P = 0.275). In BLASTp searches, HASPB sequences from rK28 had similarities with sequences from Plasmodium, encompassing software-predicted B-cell epitopes. CONCLUSIONS: The antigenicity of rK39 appears to be dependent on structural conformation, whereas that of rK28 depends on linear sequence. HASPB sequence homology with Plasmodium may be responsible for the reported cross-reactivity of rK28 with malaria sera. Further work is warranted to refine the specificity of these antigens. GRAPHICAL ABSTRACT: [Image: see text] BioMed Central 2022-10-22 /pmc/articles/PMC9587664/ /pubmed/36273150 http://dx.doi.org/10.1186/s13071-022-05495-1 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Simonson, Poppy Bhattacharyya, Tapan El-Safi, Sayda Miles, Michael A. Linear and conformational determinants of visceral leishmaniasis diagnostic antigens rK28 and rK39 |
title | Linear and conformational determinants of visceral leishmaniasis diagnostic antigens rK28 and rK39 |
title_full | Linear and conformational determinants of visceral leishmaniasis diagnostic antigens rK28 and rK39 |
title_fullStr | Linear and conformational determinants of visceral leishmaniasis diagnostic antigens rK28 and rK39 |
title_full_unstemmed | Linear and conformational determinants of visceral leishmaniasis diagnostic antigens rK28 and rK39 |
title_short | Linear and conformational determinants of visceral leishmaniasis diagnostic antigens rK28 and rK39 |
title_sort | linear and conformational determinants of visceral leishmaniasis diagnostic antigens rk28 and rk39 |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9587664/ https://www.ncbi.nlm.nih.gov/pubmed/36273150 http://dx.doi.org/10.1186/s13071-022-05495-1 |
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