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Infection and Immunity, August 2002, p. 4009-4018, Vol. 70, No. 8
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.8.4009-4018.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Salmonella enterica Serovar Typhi Live Vector Vaccines Delivered Intranasally Elicit Regional and Systemic Specific CD8+ Major Histocompatibility Class I-Restricted Cytotoxic T Lymphocytes

Marcela F. Pasetti,1* Rosangela Salerno-Gonçalves,1 and Marcelo B. Sztein1,2

Center for Vaccine Development, Department of Pediatrics,1 Department of Medicine, University of Maryland School of Medicine, Baltimore, Maryland 212012

Received 14 January 2002/ Returned for modification 11 March 2002/ Accepted 26 April 2002

We investigated the ability of live attenuated Salmonella enterica serovar Typhi strains delivered to mice intranasally to induce specific cytotoxic T-lymphocyte (CTL) responses at regional and systemic levels. Mice immunized with two doses (28 days apart) of Salmonella serovar Typhi strain Ty21a, the licensed oral typhoid vaccine, and genetically attenuated mutants CVD 908 ({Delta}aroC {Delta}aroD), CVD 915 ({Delta}guaBA), and CVD 908-htrA ({Delta}aroC {Delta}aroD {Delta}htrA) induced CTL specific for Salmonella serovar Typhi-infected cells in spleens and cervical lymph nodes. CTL were detected in effector T cells that had been expanded in vitro for 7 days in the presence of Salmonella-infected syngeneic splenocytes. A second round of stimulation further enhanced the levels of specific cytotoxicity. CTL activity was observed in sorted {alpha}ß+ CD8+ T cells, which were remarkably increased after expansion, but not in CD4+ T cells. CTL from both cervical lymph nodes and spleens failed to recognize Salmonella-infected major histocompatibility complex (MHC)-mismatched cells, indicating that the responses were MHC restricted. Studies in which MHC blocking antibodies were used showed that H-2Ld was the restriction element. This is the first demonstration that Salmonella serovar Typhi vaccines delivered intranasally elicit CD8+ MHC class I-restricted CTL. The results further support the usefulness of the murine intranasal model for evaluating the immunogenicity of typhoid vaccine candidates at the preclinical level.

* Corresponding author. Mailing address: Center for Vaccine Development, University of Maryland, 685 West Baltimore St., Room 480, Baltimore, MD 21201. Phone: (410) 706-7376. Fax: (410) 706-6205. E-mail: mpasetti{at}medicine.umaryland.edu.

Editor: J. D. Clements

Infection and Immunity, August 2002, p. 4009-4018, Vol. 70, No. 8
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.8.4009-4018.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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