Cardiac-oxidized antigens are targets of immune recognition by antibodies and potential molecular determinants in chagas disease pathogenesis

Monisha Dhiman, Maria Paola Zago, Sonia Nunez, Alejandro Amoroso, Hugo Rementeria, Pierre Dousset, Federico Nunez Burgos, Nisha Garg

Research output: Contribution to journalArticle

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Abstract

Trypanosoma cruzi elicits reactive oxygen species (ROS) of inflammatory and mitochondrial origin in infected hosts. In this study, we examined ROS-induced oxidative modifications in the heart and determined whether the resultant oxidized cardiac proteins are targets of immune response and of pathological significance in Chagas disease. Heart biopsies from chagasic mice, rats and human patients exhibited, when compared to those from normal controls, a substantial increase in protein 4-hydroxynonenal (4-HNE), malondialdehyde (MDA), carbonyl, and 3-nitrotyrosine (3-NT) adducts. To evaluate whether oxidized proteins gain antigenic properties, heart homogenates or isolated cardiomyocytes were oxidized in vitro and one- or two-dimensional gel electrophoresis (2D-GE)/Western blotting (WB) was performed to investigate the proteomic oxidative changes and recognition of oxidized proteins by sera antibodies in chagasic rodents (mice, rats) and human patients. Human cardiomyocytes exhibited LD 50 sensitivity to 30 μM 4-HNE and 100 μM H 2O 2 at 6 h and 12 h, respectively. In vitro oxidation with 4-HNE or H 2O 2 resulted in a substantial increase in 4-HNE- and carbonyl-modified proteins that correlated with increased recognition of cardiac (cardiomyocytes) proteins by sera antibodies of chagasic rodents and human patients. 2D-GE/Western blotting followed by MALDI-TOF-MS/MS analysis to identify cardiac proteins that were oxidized and recognized by human chagasic sera yielded 82 unique proteins. We validated the 2D-GE results by enzyme-linked immunosorbent assay (ELISA) and WB and demonstrated that oxidation of recombinant titin enhanced its immunogenicity and recognition by sera antibodies from chagasic hosts (rats and humans). Treatment of infected rats with phenyl-α-tert-butyl nitrone (PBN, antioxidant) resulted in normalized immune detection of cardiac proteins associated with control of cardiac pathology and preservation of heart contractile function in chagasic rats. We conclude that ROS-induced, cardiac-oxidized antigens are targets of immune recognition by antibodies and molecular determinants for pathogenesis during Chagas disease.

Original languageEnglish (US)
Article numbere28449
JournalPLoS One
Volume7
Issue number1
DOIs
StatePublished - Jan 4 2012

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Chagas disease
Chagas Disease
pathogenesis
antigens
Antigens
Rats
antibodies
Antibodies
Cardiac Myocytes
Proteins
proteins
Reactive Oxygen Species
heart
Western Blotting
rats
reactive oxygen species
Western blotting
Blood Proteins
Rodentia
Connectin

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Cardiac-oxidized antigens are targets of immune recognition by antibodies and potential molecular determinants in chagas disease pathogenesis. / Dhiman, Monisha; Zago, Maria Paola; Nunez, Sonia; Amoroso, Alejandro; Rementeria, Hugo; Dousset, Pierre; Burgos, Federico Nunez; Garg, Nisha.

In: PLoS One, Vol. 7, No. 1, e28449, 04.01.2012.

Research output: Contribution to journalArticle

Dhiman, Monisha ; Zago, Maria Paola ; Nunez, Sonia ; Amoroso, Alejandro ; Rementeria, Hugo ; Dousset, Pierre ; Burgos, Federico Nunez ; Garg, Nisha. / Cardiac-oxidized antigens are targets of immune recognition by antibodies and potential molecular determinants in chagas disease pathogenesis. In: PLoS One. 2012 ; Vol. 7, No. 1.
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AU - Amoroso, Alejandro

AU - Rementeria, Hugo

AU - Dousset, Pierre

AU - Burgos, Federico Nunez

AU - Garg, Nisha

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