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Infect Immun, August 1998, p. 3744-3751, Vol. 66, No. 8
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

A Live Recombinant Avirulent Oral Salmonella Vaccine Expressing Pneumococcal Surface Protein A Induces Protective Responses against Streptococcus pneumoniae

Amiya R. Nayak,1 Steven A. Tinge,1 2 Rebecca C. Tart,3 dagger Larry S. McDaniel,3 Dagger David E. Briles,3 and Roy Curtiss III1 *

Department of Biology, Washington University, St. Louis, Missouri 631301; MEGAN Health Incorporated, St. Louis, Missouri 631102; and Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 352943

Received 7 January 1998/Returned for modification 9 February 1998/Accepted 31 May 1998

A live oral recombinant Salmonella vaccine strain expressing pneumococcal surface protein A (PspA) was developed. The strain was attenuated with Delta cya Delta crp mutations. Stable expression of PspA was achieved by the use of the balanced-lethal vector-host system, which employs an asd deletion in the host chromosome to impose an obligate requirement for diaminopimelic acid. The chromosomal Delta asd mutation was complemented by a plasmid vector possessing the asd+ gene. A portion of the pspA gene from Streptococcus pneumoniae Rx1 was cloned onto a multicopy Asd+ vector. After oral immunization, the recombinant Salmonella-PspA vaccine strain colonized the Peyer's patches, spleens, and livers of BALB/cByJ and CBA/N mice and stimulated humoral and mucosal antibody responses. Oral immunization of outbred New Zealand White rabbits with the recombinant Salmonella strain induced significant anti-PspA immunoglobulin G titers in serum and vaginal secretions. Polyclonal sera from orally immunized mice detected PspA on the S. pneumoniae cell surface as revealed by immunofluorescence. Oral immunization of BALB/cJ mice with the PspA-producing Salmonella strain elicited antibody to PspA and resistance to challenge by the mouse-virulent human clinical isolate S. pneumoniae WU2. Immune sera from orally immunized mice conferred passive protection against otherwise lethal intraperitoneal or intravascular challenge with strain WU2.

* Corresponding author. Mailing address: Department of Biology, Washington University, Campus Box 1137, One Brookings Dr., St. Louis, MO 63130-4899. Phone: (314) 935-6819. Fax: (314) 935-7246. E-mail: kvatern{at}biodec.wustl.edu.

dagger Present address: Department of Biological Sciences, Campbell University, Buies Creek, NC 27506.

Dagger Present address: Departments of Surgery and Microbiology, School of Medicine, The University of Mississippi Medical Center, Jackson, MS 39126.

Infect Immun, August 1998, p. 3744-3751, Vol. 66, No. 8
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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