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Infection and Immunity, July 2003, p. 3971-3978, Vol. 71, No. 7
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.7.3971-3978.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Development of DNA Vaccines against Hemolytic-Uremic Syndrome in a Murine Model

Alejandra V. E. Capozzo,1 Virginia Pistone Creydt,2 Graciela Dran,1 Gabriela Fernández,1 Sonia Gómez,1 Leticia V. Bentancor,1 Carolina Rubel,1 Cristina Ibarra,2 Martín Isturiz,1 and Marina S. Palermo1*

Instituto de Investigaciones Hematológicas, Academia Nacional de Medicina,1 Departamento de Fisiología, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina2

Received 9 October 2002/ Returned for modification 26 November 2002/ Accepted 12 March 2003

Shiga toxin type 2 (Stx2) produced by Escherichia coli O:157H7 can cause hemolytic-uremic syndrome in children, a disease for which there is neither a vaccine nor an effective treatment. This toxin consists of an enzymatically active A subunit and a pentameric B subunit responsible for the toxin binding to host cells, and also found to be immunogenic in rabbits. In this study we developed eukaryotic plasmids expressing the B subunit gene of Stx2 (pStx2B) and the B subunit plus the gene coding for the A subunit with an active-site deletion (pStx2{Delta}A). Transfection of eukaryotic cells with these plasmids produced proteins of the expected molecular weight which reacted with specific monoclonal antibodies. Newborn and adult BALB/c mice immunized with two intramuscular injections of each plasmid, either alone or together with the same vector expressing the granulocyte and monocyte colony-stimulating factor (pGM-CSF), elicited a specific Th1-biased humoral response. The effect of pGM-CSF as an adjuvant plasmid was particularly notable in newborn mice and in pStx2B-vaccinated adult mice. Stx2-neutralizing activity, evaluated in vitro on VERO cell monolayers, correlated with in vivo protection. This is the first report using plasmids to induce a neutralizing humoral immune response against the Stx2.

* Corresponding author. Mailing address: Div. Inmunología, Instituto de Investigaciones Hematológicas, Academia Nacional de Medicina, P. de Melo 3081 (1425), Buenos Aires, Argentina. Phone: 5411-4805-5695. Fax: 5411-4803-9475. E-mail: mspalermo{at}hematologia.anm.edu.ar.

Editor: A. D. O'Brien

Infection and Immunity, July 2003, p. 3971-3978, Vol. 71, No. 7
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.7.3971-3978.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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