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Comparative Study
. 2002 Sep 10;20(27-28):3322-31.
doi: 10.1016/s0264-410x(02)00310-9.

Comparative study of immune responses induced after immunization with plasmids encoding the HIV-1 Nef protein under the control of the CMV-IE or the muscle-specific desmin promoter

Nicolas Cazeaux  1 , Yamina Bennasser, Pierre-Louis Vidal, Zhenlin Li, Denise Paulin, Elmostafa Bahraoui
Affiliations
Comparative Study

Comparative study of immune responses induced after immunization with plasmids encoding the HIV-1 Nef protein under the control of the CMV-IE or the muscle-specific desmin promoter

Nicolas Cazeaux et al. Vaccine. .

Abstract

The objective of this study was to compare immune responses induced against HIV-1 Nef after DNA immunization with Nef encoding plasmids under the control of an ubiquitous or a muscle-specific promoter. To this end, plasmids containing HIV-1 nef under the control of the Cytomegalovirus or the human desmin promoters, specifically expressed in muscle cells, were constructed. Different groups of BALB/c mice were immunized with 10 or 100 microg of both constructs, controls were the pcDNA3 vector or the Nef protein in Freund's adjuvant (Nef-CFA). Our data showed that both plasmids stimulated anti-Nef humoral responses in a dose-dependent manner. The anti-Nef antibody response, however, was earlier and higher with the CMV-IE promoter than with the desmin promoter. We also showed that Nef expressing plasmids induced high titers of anti-Nef antibodies (10(4)), comparable to those obtained in the Nef-CFA group (4x10(4)). Analysis of the specificity of anti-Nef antibodies revealed no influence of the promoter, and in contrast to Nef-CFA, plasmid immunization elicited anti-Nef antibodies directed principally against conformational-dependent epitopes. Data on the lymphoproliferative response showed that the specificity of expression, or the plasmid dose, did not affect the onset-time or the intensity of the response. The predominant IgG2a isotype of anti-Nef antibodies and the cytokine profile, mostly IL-2 and IFN-gamma, produced by Nef-stimulated spleen cells indicated a Th1 response in plasmid-immunized mice, in contrast to mice immunized with Nef-CFA, where a Th2 response was induced. In conclusion, these data indicate that antigen expression by muscle cells is sufficient to stimulate a Th1 immune response.

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