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Journal of Virology, June 2003, p. 7048-7057, Vol. 77, No. 12
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.12.7048-7057.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Prime-Boost Immunization Schedules Based on Influenza Virus and Vaccinia Virus Vectors Potentiate Cellular Immune Responses against Human Immunodeficiency Virus Env Protein Systemically and in the Genitorectal Draining Lymph Nodes

M. Magdalena Gherardi,1 José Luis Nájera,1 Eva Pérez-Jiménez,1 Susana Guerra,1 Adolfo García-Sastre,2 and Mariano Esteban1*

Department of Molecular and Cellular Biology, Centro Nacional de Biotecnologia, CSIC, Campus Universidad Autónoma, 28049 Madrid, Spain,1 Department of Microbiology, Mount Sinai School of Medicine, New York, New York 100292

Received 20 December 2002/ Accepted 15 March 2003

Vaccines that elicit systemic and mucosal immune responses should be the choice to control human immunodeficiency virus (HIV) infections. We have previously shown that prime-boost immunizations with influenza virus Env and vaccinia virus (VV) WR Env recombinants induced an enhanced systemic CD8+ T-cell response against HIV-1 Env antigen. In this report, we analyzed in BALB/c mice after priming with influenza virus Env the ability of two VV recombinants expressing HIV-1 Env B (VV WR Env and the highly attenuated modified VV Ankara [MVA] Env) to boost cellular immune responses in the spleen and in the lymph nodes draining the genital and rectal tracts. Groups of mice were primed by the intranasal route with 104 PFU of influenza virus Env and boosted 14 days later by the intraperitoneal or intranasal route with 107 PFU of MVA Env or VV WR Env, while the control group received two immunizations with influenza virus Env. We found that the combined immunization (Flu/VV) increased more than 60 times the number of gamma interferon-specific CD8+ T cells compared to the Flu/Flu scheme. Significantly, boosting with MVA Env by the intraperitoneal route induced a response 1.25 or 2.5 times (spleen or genital lymph nodes) higher with respect to that found after the boost with VV WR Env. Mice with an enhanced CD8+ T-cell response also had an increased Th1/Th2 ratio, evaluated by the cytokine pattern secreted following in vitro restimulation with gp160 protein and by the specific immunoglobulin G2a (IgG2a)/IgG1 ratio in serum. By the intranasal route recombinant WR Env booster gave a more efficient immune response (10 and 1.3 times in spleen and genital lymph nodes, respectively) than recombinant MVA Env. However, the scheme influenza virus Env/MVA Env increased four times the response in the spleen, giving a low but significant response in the genital lymph nodes compared with a single intranasal immunization with MVA Env. These results demonstrate that the combination Flu/MVA in prime-booster immunization regimens is an effective vaccination approach to generate cellular immune responses to HIV antigens at sites critical for protective responses.

* Corresponding author. Mailing address: Centro Nacional de Biotecnología, Campus Universidad Autónoma, 28049 Madrid, Spain. Phone: 34 91 585 4506. Fax: 34 91 585 4506. E-mail: mesteban{at}cnb.uam.es.

Journal of Virology, June 2003, p. 7048-7057, Vol. 77, No. 12
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.12.7048-7057.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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