Recombinant antibody against Trypanosoma cruzi from patients with chronic Chagas heart disease recognizes mammalian nervous system.
Amino Acid Sequence
Animals
Antibodies, Monoclonal
/ immunology
Antibodies, Protozoan
/ immunology
Antibody Specificity
/ immunology
Antigens, Protozoan
/ immunology
Cell Line
Chagas Disease
/ drug therapy
Cloning, Molecular
Cross Reactions
/ immunology
Drug Development
Enzyme-Linked Immunosorbent Assay
Escherichia coli
/ genetics
Fluorescent Antibody Technique
Gene Expression
Humans
Immunoprecipitation
Mass Spectrometry
Mice
Molecular Mimicry
Rats
Recombinant Fusion Proteins
/ isolation & purification
Sequence Analysis, DNA
Single-Chain Antibodies
/ immunology
Trypanosoma cruzi
/ immunology
Chagas disease
Digestive system
Molecular mimicry
Phage-display
Post-translational modification
Tubulin
Journal
EBioMedicine
ISSN: 2352-3964
Titre abrégé: EBioMedicine
Pays: Netherlands
ID NLM: 101647039
Informations de publication
Date de publication:
Jan 2021
Jan 2021
Historique:
received:
08
09
2020
revised:
15
10
2020
accepted:
26
11
2020
pubmed:
12
1
2021
medline:
6
10
2021
entrez:
11
1
2021
Statut:
ppublish
Résumé
To deeply understand the role of antibodies in the context of Trypanosoma cruzi infection, we decided to characterize A2R1, a parasite antibody selected from single-chain variable fragment (scFv) phage display libraries constructed from B cells of chronic Chagas heart disease patients. Immunoblot, ELISA, cytometry, immunofluorescence and immunohistochemical assays were used to characterize A2R1 reactivity. To identify the antibody target, we performed an immunoprecipitation and two-dimensional electrophoresis coupled to mass spectrometry and confirmed A2R1 specific interaction by producing the antigen in different expression systems. Based on these data, we carried out a comparative in silico analysis of the protein target´s orthologues, focusing mainly on post-translational modifications. A2R1 recognizes a parasite protein of ~50 kDa present in all life cycle stages of T. cruzi, as well as in other members of the kinetoplastid family, showing a defined immunofluorescence labeling pattern consistent with the cytoskeleton. A2R1 binds to tubulin, but this interaction relies on its post-translational modifications. Interestingly, this antibody also targets mammalian tubulin only present in brain, staining in and around cell bodies of the human peripheral and central nervous system. Our findings demonstrate for the first time the existence of a human antibody against T. cruzi tubulin capable of cross-reacting with a human neural protein. This work re-emphasizes the role of molecular mimicry between host and parasitic antigens in the development of pathological manifestations of T. cruzi infection.
Sections du résumé
BACKGROUND
BACKGROUND
To deeply understand the role of antibodies in the context of Trypanosoma cruzi infection, we decided to characterize A2R1, a parasite antibody selected from single-chain variable fragment (scFv) phage display libraries constructed from B cells of chronic Chagas heart disease patients.
METHODS
METHODS
Immunoblot, ELISA, cytometry, immunofluorescence and immunohistochemical assays were used to characterize A2R1 reactivity. To identify the antibody target, we performed an immunoprecipitation and two-dimensional electrophoresis coupled to mass spectrometry and confirmed A2R1 specific interaction by producing the antigen in different expression systems. Based on these data, we carried out a comparative in silico analysis of the protein target´s orthologues, focusing mainly on post-translational modifications.
FINDINGS
RESULTS
A2R1 recognizes a parasite protein of ~50 kDa present in all life cycle stages of T. cruzi, as well as in other members of the kinetoplastid family, showing a defined immunofluorescence labeling pattern consistent with the cytoskeleton. A2R1 binds to tubulin, but this interaction relies on its post-translational modifications. Interestingly, this antibody also targets mammalian tubulin only present in brain, staining in and around cell bodies of the human peripheral and central nervous system.
INTERPRETATION
CONCLUSIONS
Our findings demonstrate for the first time the existence of a human antibody against T. cruzi tubulin capable of cross-reacting with a human neural protein. This work re-emphasizes the role of molecular mimicry between host and parasitic antigens in the development of pathological manifestations of T. cruzi infection.
Identifiants
pubmed: 33429173
pii: S2352-3964(20)30582-X
doi: 10.1016/j.ebiom.2020.103206
pmc: PMC7809174
pii:
doi:
Substances chimiques
Antibodies, Monoclonal
0
Antibodies, Protozoan
0
Antigens, Protozoan
0
Recombinant Fusion Proteins
0
Single-Chain Antibodies
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
103206Informations de copyright
Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Competing Interests The authors have declared that no conflict of interest exists.
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