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Human recombinant antibodies against Trypanosoma cruzi ribosomal P2β protein

Published online by Cambridge University Press:  18 March 2011

VANINA GRIPPO ,
LETICIA L. NIBORSKI ,
KARINA A. GOMEZ  and
MARIANO J. LEVIN
VANINA GRIPPO
Affiliation:
Laboratory of Molecular Biology of Chagas' Disease, Institute for Genetic Engineering and Molecular Biology (INGEBI), CONICET-UBA, 1428 Ciudad autónoma de Buenos Aires, Argentina
LETICIA L. NIBORSKI
Affiliation:
Laboratory of Molecular Biology of Chagas' Disease, Institute for Genetic Engineering and Molecular Biology (INGEBI), CONICET-UBA, 1428 Ciudad autónoma de Buenos Aires, Argentina
KARINA A. GOMEZ*
Affiliation:
Laboratory of Molecular Biology of Chagas' Disease, Institute for Genetic Engineering and Molecular Biology (INGEBI), CONICET-UBA, 1428 Ciudad autónoma de Buenos Aires, Argentina
MARIANO J. LEVIN
Affiliation:
Laboratory of Molecular Biology of Chagas' Disease, Institute for Genetic Engineering and Molecular Biology (INGEBI), CONICET-UBA, 1428 Ciudad autónoma de Buenos Aires, Argentina
*
*Corresponding author: Laboratorio de Biología Molecular de la Enfermedad de Chagas, INGEBI. Vuelta de Obligado 2490, 1428-Ciudad autónoma de Buenos Aires-Argentina. Tel: 00 54 11 47832871. Fax: 00 54 11 47868578. E-mail: [email protected]
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Summary

Patients with chronic Chagas' Heart Disease (cChHD) develop an antibody response that is suspected to be involved in the cardiac pathogenesis. The response against Trypanosoma cruzi ribosomal P proteins is of particular interest, as these antibodies can cross-react with host cardiac receptors causing electrophysiological alterations. To better understand the humoral anti-P response we constructed a single-chain variable fragment library derived from a cChHD patient. The variable heavy and light regions were amplified from bone-marrow RNA and subcloned into the vector pComb3X. The phage library was subsequently panned against T. cruzi ribosomal P2β protein (TcP2β). We obtained 3 different human recombinant antibodies that specifically reacted with TcP2β in ELISA and Western blots. Two of them reacted with the C-terminal region of TcP2β, peptide R13, as the recombinant autoanti-P antibodies from Systemic Lupus Erythematosus (SLE) patients. Interestingly, the third one was specific for TcP2β but did not recognize R13, confirming the specific nature of the anti-P response in Chagas disease. Neither sequence nor VH usage similarities between Chagas and SLE anti-P autoantibodies were observed. Herein, the first human mAbs against TcP2β have been obtained and characterized showing that the humoral anti-P response is directed against the parasite and does not include an autoimmune component.

Keywords

phage displayhuman recombinant antibodiesribosomal P proteinChagas diseaseTrypanosoma cruzi
Type
Research Article
Information
Parasitology , Volume 138 , Issue 6 , May 2011 , pp. 736 - 747
Copyright
Copyright © Cambridge University Press 2011

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