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Infection and Immunity, June 2003, p. 3165-3171, Vol. 71, No. 6
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.6.3165-3171.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Humoral and Cellular Immune Responses to Trypanosoma cruzi-Derived Paraflagellar Rod Proteins in Patients with Chagas' Disease

Vladimir Michailowsky,1 Keith Luhrs,2 Manoel Otávio C. Rocha,3 David Fouts,2 Ricardo T. Gazzinelli,1,4* and Jerry E. Manning2*

Laboratory of Immunopathology, René Rachou Research Center—Oswaldo Cruz Foundation,1 Department of Biochemistry and Immunology, Institute of Biological Sciences,4 Faculty of Medicine, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais, Brazil,3 Department of Molecular Biology and Biochemistry, University of California, Irvine, California2

Received 9 September 2002/ Returned for modification 5 December 2002/ Accepted 28 February 2003

Sera and peripheral blood mononuclear cells (PBMC) from patients displaying different clinical symptoms as well as from normal uninfected individuals (NI) were used to evaluate the humoral and cellular responses of Chagas' disease patients to Trypanosoma cruzi-derived paraflagellar rod proteins (PFR). Our results show that sera from both asymptomatic Chagas' disease patients (ACP) and cardiac Chagas' disease patients (CCP) have higher levels of antibodies to PFR than sera from NI. Immunoglobulin G1 (IgG1) and IgG3 were the main Ig isotypes that recognized PFR. We also tested three recombinant forms of PFR, named rPAR-1, rPAR-2, and rPAR-3, by Western blot analysis. Sera from seven out of eight patients with Chagas' disease recognized one of the three rPAR forms. Sera from 75, 50, and 37.5% of Chagas' disease patients tested recognized rPAR-3, rPAR-2, and rPAR-1, respectively. PFR induced proliferation of 100 and 70% of PBMC from ACP and CCP, respectively. Further, stimulation of cells from Chagas' disease patients with PFR enhanced the frequencies of both small and large CD4+ CD25+ and CD4+ CD69+ lymphocytes, as well as that of small CD8+ CD25+ lymphocytes. Finally, we evaluated the ability of PFR to elicit the production of gamma interferon (IFN-{gamma}) by PBMC from patients with Chagas' disease. Fifty percent of the PBMC from ACP as well as CCP produced IFN-{gamma} upon stimulation with PFR. PFR enhanced the percentages of IFN-{gamma}-producing cells in both CD3+ and CD3- populations. Within the T-cell population, large CD4+ T lymphocytes were the main source of IFN-{gamma}.

* Corresponding author. Mailing address for Jerry E. Manning: Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92717. Phone: (949) 824-5578. Fax: (949) 824-8551. E-mail: jemanning@uci.edu. Mailing address for Ricardo T. Gazzinelli: Department of Biochemistry and Immunology, Federal University of Minas Gerais, Av. Antônio Carlos 6627, 31270-910 Belo Horizonte, Minas Gerais, Brazil. Phone: 0055 31 295 3566. Fax: 0055 31 295 3115. E-mail: ritoga{at}dedalus.lcc.ufmg.br.

Editor: J. M. Mansfield

Infection and Immunity, June 2003, p. 3165-3171, Vol. 71, No. 6
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.6.3165-3171.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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