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Clinical and Diagnostic Laboratory Immunology, July 2004, p. 704-710, Vol. 11, No. 4
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.4.704-710.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Recombinant GRA4 or ROP2 Protein Combined with Alum or the gra4 Gene Provides Partial Protection in Chronic Murine Models of Toxoplasmosis

Valentina Martin,1 Alicia Supanitsky,1 Pablo C. Echeverria,1,2 Silvana Litwin,3 Tamara Tanos,1 Adolfo R. De Roodt,3 Eduardo A. Guarnera,1 and Sergio O. Angel1,2*

Departamento de Parasitología, Instituto Nacional de Enfermedades Infecciosas,1 Instituto Nacional de Producción de Biológicos, ANLIS Dr. Carlos G. Malbran, Ciudad de Buenos Aires,3 Laboratorio de Parasitología Molecular, UB2, IIB-INTECH, CONICET-UNSAM, Chascomús, Provincia Buenos Aires, Argentina2

Received 9 January 2004/ Returned for modification 17 February 2004/ Accepted 7 April 2004

The efficacy of vaccination with Toxoplasma gondii recombinant GRA4 (rGRA4) and ROP2 (rRPO2) proteins and a mix of both combined with alum were evaluated in C57BL/6 and C3H mice. In C57BL/6 mice, rGRA4 and rGRA4-rROP2 immunizations generated similar levels of immunoglobulin G1 (IgG1) and IgG2a isotypes against GRA4, whereas immunizations with rROP2 and the mix induced a predominant IgG1 production against ROP2. All groups of C3H vaccinated mice exhibited higher levels of IgG1 than IgG2a. rGRA4-stimulated splenocytes from vaccinated mice produced primarily gamma interferon while those stimulated with rROP2 produced interleukin-4. Challenge of rGRA4- or rGRA4-rROP2-vaccinated mice from both strains with ME49 cysts resulted in fewer brain cysts than the controls, whereas vaccination with rROP2 alone only conferred protection to C3H mice. Immunization with a plasmid carrying the entire open reading frame of GRA4 showed a protective level similar to that of rGRA4 combined with alum. These results suggest that GRA4 can be a good candidate for a multiantigen anti-T. gondii vaccine based on the use of alum as an adjuvant.

* Corresponding author. Mailing address: Laboratorio de Parasitología Molecular, UB2, IIB-INTECH, CONICET-UNSAM, Camino de Circunvalación Laguna Km. 6, C.C. 164 (B7130IIWA), Chascomús, Prov. Buenos Aires, Argentina. Phone: 54 2241 430323. Fax: 54 2241 424048. E-mail: sangel{at}intech.gov.ar.

Clinical and Diagnostic Laboratory Immunology, July 2004, p. 704-710, Vol. 11, No. 4
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.4.704-710.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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