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Infection and Immunity, January 2000, p. 205-213, Vol. 68, No. 1
0019-9567/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

In Vivo Characterization of the Murine Intranasal Model for Assessing the Immunogenicity of Attenuated Salmonella enterica Serovar Typhi Strains as Live Mucosal Vaccines and as Live Vectors

Thames E. Pickett,1,2 Marcela F. Pasetti,1 James E. Galen,1 Marcelo B. Sztein,1,2 and Myron M. Levine1,2,*

Center for Vaccine Development, Division of Geographic Medicine, Department of Medicine, and Division of Infectious Diseases and Tropical Pediatrics, Department of Pediatrics,1 and Department of Microbiology and Immunology,2 University of Maryland School of Medicine, Baltimore, Maryland 21201

Received 27 July 1999/Returned for modification 13 September 1999/Accepted 5 October 1999

Attenuated Salmonella enterica serovar Typhi live vector vaccine strains are highly immunogenic in mice following intranasal but not orogastric inoculation. To elucidate the relationship between organs within which vaccine organisms are found and the induction of specific serum immunoglobulin G (IgG) antibodies, we examined the in vivo distribution of serovar Typhi vaccine strain CVD 908-htrA following intranasal administration. Vaccine organisms were cultured from the nasal lymphoid tissue (NALT), lungs, and Peyer's patches 2 min after intranasal inoculation. Vaccine organisms persisted longer in NALT than in other organs. By decreasing the volume of intranasal inoculum containing 109 CFU (from a single 30- or 10-µl dose to four 2.5-µl doses given over the course of 1 h), we were able to significantly reduce the number of vaccine organisms isolated from the lungs (P < 0.05) without reducing the number of vaccine organisms in NALT. Reducing the number of vaccine organisms in the lungs resulted in a significant decrease in the serum tetanus antitoxin response elicited by CVD 908-htrA expressing tetanus toxin fragment C under the control of the redox-responsive nir15 promoter. In contrast, a similar construct expressing tetanus toxin fragment C under control of the constitutive lpp promoter stimulated a strong serum IgG tetanus antitoxin response with both inoculation regimens. The data suggest that following intranasal inoculation, NALT is a sufficient inductive site for elicitation of an immune response against both the live vector and heterologous antigen and, as occurs following oral inoculation of humans, attenuated serovar Typhi vaccine organisms elicit serum IgG responses.

* Corresponding author. Mailing address: Center for Vaccine Development, University of Maryland School of Medicine, HSF Rm. 480, 685 W. Baltimore St., Baltimore, MD 21201. Phone: (410) 706-2493. Fax: (410) 706-6205. E-mail: mlevine{at}umppa1.ab.umd.edu.

Infection and Immunity, January 2000, p. 205-213, Vol. 68, No. 1
0019-9567/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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