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Journal of Virology, May 2003, p. 5209-5217, Vol. 77, No. 9
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.9.5209-5217.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Attenuated Salmonella enterica Serovar Typhi and Shigella flexneri 2a Strains Mucosally Deliver DNA Vaccines Encoding Measles Virus Hemagglutinin, Inducing Specific Immune Responses and Protection in Cotton Rats

Marcela F. Pasetti,1,2 Eileen M. Barry,1,3 Genevieve Losonsky,1,2 Mahender Singh,1 Sandra M. Medina-Moreno,1,3 John M. Polo,4 Jeffrey Ulmer,4 Harriet Robinson,5 Marcelo B. Sztein,1,2,3 and Myron M. Levine1,2,3*

Center for Vaccine Development,1 Departments of Pediatrics,2 Medicine, University of Maryland School of Medicine, Baltimore, Maryland,3 Chiron Vaccines, Emeryville, California,4 Vaccine Center, Emory University, Atlanta, Georgia5

Received 21 October 2002/ Accepted 27 January 2003

Measles remains a leading cause of child mortality in developing countries. Residual maternal measles antibodies and immunologic immaturity dampen immunogenicity of the current vaccine in young infants. Because cotton rat respiratory tract is susceptible to measles virus (MV) replication after intranasal (i.n.) challenge, this model can be used to assess the efficacy of MV vaccines. Pursuing a new measles vaccine strategy that might be effective in young infants, we used attenuated Salmonella enterica serovar Typhi CVD 908-htrA and Shigella flexneri 2a CVD 1208 vaccines to deliver mucosally to cotton rats eukaryotic expression plasmid pGA3-mH and Sindbis virus-based DNA replicon pMSIN-H encoding MV hemagglutinin (H). The initial i.n. dose-response with bacterial vectors alone identified a well-tolerated dosage (1 x 109 to 7 x 109 CFU) and a volume (20 µl) that elicited strong antivector immune responses. Animals immunized i.n. on days 0, 28, and 76 with bacterial vectors carrying DNA plasmids encoding MV H or immunized parenterally with these naked DNA vaccine plasmids developed MV plaque reduction neutralizing antibodies and proliferative responses against MV antigens. In a subsequent experiment of identical design, cotton rats were challenged with wild-type MV 1 month after the third dose of vaccine or placebo. MV titers were significantly reduced in lung tissue of animals immunized with MV DNA vaccines delivered either via bacterial live vectors or parenterally. Since attenuated serovar Typhi and S. flexneri can deliver measles DNA vaccines mucosally in cotton rats, inducing measles immune responses (including neutralizing antibodies) and protection, boosting strategies can now be evaluated in animals primed with MV DNA vaccines.

* Corresponding author. Mailing address: Center for Vaccine Development, University of Maryland School of Medicine, 685 W. Baltimore St., Rm. 480, Baltimore, MD 21201. Phone: (410) 706-7588. Fax: (410) 706-6205. E-mail: mlevine{at}medicine.umaryland.edu.

Journal of Virology, May 2003, p. 5209-5217, Vol. 77, No. 9
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.9.5209-5217.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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