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Am. J. Trop. Med. Hyg., 69(2), 2003, pp. 129-134
Copyright © 2003 by The American Society of Tropical Medicine and Hygiene

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INDUCTION OF NEUTRALIZING ANTIBODIES AND PARTIAL PROTECTION FROM VIRAL CHALLENGE IN MACACA FASCICULARIS IMMUNIZED WITH RECOMBINANT DENGUE 4 VIRUS ENVELOPE GLYCOPROTEIN EXPRESSED IN PICHIA PASTORIS

MARÍA G. GUZMÁN, RAYNER RODRÍGUEZ, ROSMARI RODRÍGUEZ, LISSET HERMIDA, MAYLING ALVAREZ, LAURA LAZO, MAYRA MUNÉ, DELFINA ROSARIO, KATIA VALDÉS, SUSANA VÁZQUEZ, RAFAEL MARTINEZ, TERESITA SERRANO, JORGE PAEZ, RAÚL ESPINOSA, TANIA PUMARIEGA, AND GERARDO GUILLÉN
Virology Department, Pan American Health Organization/World Health Organization Collaborating Center for Viral Diseases, Pedro Kourí Tropical Medicine Institute, Autopista Novia del Mediodía, Havana, Cuba; Genetic Engineering and Biotechnology Center, Havana, Cuba; Latin American School of Medicine, Havana, Cuba

A recombinant vaccine that expresses the envelope (E) gene of dengue virus type 4 was tested for immunogenicity and protection in Macaca fascicularis. One hundred micrograms of semipurified recombinant E protein (E4rec) expressed in Pichia pastoris was used to immunize three animals. Neutralizing antibodies to dengue 4 virus with a titer of 1:30 were detected in all immunized monkeys prior to challenge. Animals were challenged with 105 plaque-forming units of dengue 4 virus. One vaccine-immunized monkey was protected from viremia, while the other two were partially protected. Monkeys immunized with E4rec elicited the highest neutralizing antibody titers (P < 0.05) ranging from 1:85 to 1:640 at day 30. In both immunized and control animals, the longest duration of viremia correlated with earliest and highest level of IgM antibody to dengue virus. The vaccinated animals showed anamnestic antibody responses upon virus challenge, indicating successful priming by the recombinant vaccine. Our results suggest that E4rec expressed in P. pastoris can provide partial protection against viremia. However, the results were not effective enough to use it as a vaccine candidate. Further work is required to improve the quality of the immunogen.

Received September 30, 2002. Accepted for publication April 12, 2003.

Acknowledgments: We thank Drs. Roberto Fernández and Omar Fuentes for useful comments regarding biosafety and entomologic control, María E. Moreno for technical assistance, Dr. Karelia Cozme for contributions in the animal studies, and Drs. Robert Shope and John David for useful suggestions and comments.

Financial support: This study was supported by the Cuban Program for Vaccine Development.

Authors’ addresses: María G. Guzmán, Rayner Rodríguez, Rosmari Rodríguez, Mayling Alvarez, Mayra Muné, Delfina Rosario, Susana Vázquez, Teresita Serrano, and Tania Pumariega, Pedro Kourí Tropical Medicine Institute, Autopista Novia del Mediodía, Km 6, PO Box Marianao 13, Havana, Cuba, Telephone: 53-7-202-0450 and 53-7-202-0633, Fax: 53-7-204-6051, E-mail: lupe{at}ipk.sld.cu. Lisset Hermida, Laura Lazo, Rafael Martinez, Jorge Paez, Raúl Espinosa, and Gerardo Guillén, Genetic Engineering and Biotechnology Center, Apartado 6162, Havana, Cuba. Katia Valdés, Latin American School of Medicine, Havana, Cuba.




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Copyright © 2003 by the American Society of Tropical Medicine and Hygiene.