Development of ELISA based on Bacillus anthracis capsule biosynthesis protein CapA for naturally acquired antibodies against anthrax.
Animals
Anthrax
/ immunology
Anthrax Vaccines
/ immunology
Antibodies, Bacterial
/ immunology
Antigens, Bacterial
/ immunology
Bacillus anthracis
/ immunology
Bacterial Capsules
/ immunology
Bacterial Proteins
/ immunology
Enzyme-Linked Immunosorbent Assay
/ methods
Heat-Shock Proteins
/ immunology
Horses
Immunoglobulin G
/ immunology
Plasmids
/ metabolism
Sequence Homology, Amino Acid
Spores, Bacterial
/ immunology
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2021
2021
Historique:
received:
18
06
2021
accepted:
23
09
2021
entrez:
11
10
2021
pubmed:
12
10
2021
medline:
26
11
2021
Statut:
epublish
Résumé
Anthrax is a zoonotic disease caused by the gram-positive spore-forming bacterium Bacillus anthracis. Detecting naturally acquired antibodies against anthrax sublethal exposure in animals is essential for anthrax surveillance and effective control measures. Serological assays based on protective antigen (PA) of B. anthracis are mainly used for anthrax surveillance and vaccine evaluation. Although the assay is reliable, it is challenging to distinguish the naturally acquired antibodies from vaccine-induced immunity in animals because PA is cross-reactive to both antibodies. Although additional data on the vaccination history of animals could bypass this problem, such data are not readily accessible in many cases. In this study, we established a new enzyme-linked immunosorbent assay (ELISA) specific to antibodies against capsule biosynthesis protein CapA antigen of B. anthracis, which is non-cross-reactive to vaccine-induced antibodies in horses. Using in silico analyses, we screened coding sequences encoded on pXO2 plasmid, which is absent in the veterinary vaccine strain Sterne 34F2 but present in virulent strains of B. anthracis. Among the 8 selected antigen candidates, capsule biosynthesis protein CapA (GBAA_RS28240) and peptide ABC transporter substrate-binding protein (GBAA_RS28340) were detected by antibodies in infected horse sera. Of these, CapA has not yet been identified as immunoreactive in other studies to the best of our knowledge. Considering the protein solubility and specificity of B. anthracis, we prepared the C-terminus region of CapA, named CapA322, and developed CapA322-ELISA based on a horse model. Comparative analysis of the CapA322-ELISA and PAD1-ELISA (ELISA uses domain one of the PA) showed that CapA322-ELISA could detect anti-CapA antibodies in sera from infected horses but was non-reactive to sera from vaccinated horses. The CapA322-ELISA could contribute to the anthrax surveillance in endemic areas, and two immunoreactive proteins identified in this study could be additives to the improvement of current or future vaccine development.
Identifiants
pubmed: 34634075
doi: 10.1371/journal.pone.0258317
pii: PONE-D-21-20020
pmc: PMC8504768
doi:
Substances chimiques
Anthrax Vaccines
0
Antibodies, Bacterial
0
Antigens, Bacterial
0
Bacterial Proteins
0
CapA protein, bacteria
0
Heat-Shock Proteins
0
Immunoglobulin G
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0258317Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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