Identification of proteins recognized by immune sera from chagasic patients.

Human cardiomyocyte lysates were treated with 100-µM H2O2 or 30-µM 4-hydroxynonenal (4-HNE). Cell lysates were resolved by 2 dimensional gel electrophoresis and probed with sera samples from normal subjects, seropositive chagasic patients that exhibited no clinical disease (CD0–1) or mild-moderate symptoms of clinical disease (CD2–3), and seronegative other cardiomyopathy (OCM) patients. Protein spots recognized by immune sera were submitted to MALDI-TOF-MS/MS analysis. A −, +, ++, and +++ indicates none, mild, moderate and strong recognition of protein spots by immune sera. Gels were also probed with anti-dinitrophenyl (DNP) and anti-4-HNE antibodies. In column 1, * and # indicate protein spots that were recognized by anti-HNE and anti-DNP antibodies, respectively.