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Infection and Immunity, March 2002, p. 1434-1442, Vol. 70, No. 3
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.3.1434-1442.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Extracellular Antigens from Salmonella enteritidis Induce Effective Immune Response in Mice after Oral Vaccination

Lena Strindelius,1 Lena Degling Wikingsson,1,2 and Ingvar Sjöholm1*

Department of Pharmacy, Biomedical Center, Uppsala University, SE-751 23 Uppsala ,1 SBL Vaccin AB, SE-105 21 Stockholm, Sweden2

Received 4 September 2001/ Accepted 30 November 2001

We have studied polyacryl starch microparticles as an adjuvant in oral vaccination in mice. Secreted antigens from Salmonella enterica serovar Enteritidis were administered covalently conjugated to microparticles, or as free antigens, orally or intramuscularly and evaluated for their immunogenicity and ability to elicit protective immune response against an oral challenge with live serovar Enteritidis. The highest immunoglobulin M (IgM)-plus-IgG titers were obtained in the groups immunized with antigen-conjugated microparticles. The subclass profile switched to a stronger Th1 influence in the oral groups after booster, while the intramuscular group showed a constant Th1/Th2 profile. A strong specific IgA response was seen in feces in the oral groups, which was further confirmed in an enzyme-linked immunospot assay. The delayed-type hypersensitivity test, as a measure of the cellular response, showed a significant increase in ear thickness in all the immunized groups, except for the group that received free antigen orally, compared to the nonimmunized group. The cytokines released from in vitro-stimulated spleens showed a strong gamma interferon response in all immunized groups. A significant reduction in CFU in liver and spleen was seen in the orally immunized groups compared to the nonimmunized group after oral challenge with serovar Enteritidis. Western blotting analysis with both sera and feces revealed that antibodies against three bands, 53, 56, and 60 kDa, dominated the oral groups, and an electrospray-mass spectroscopy analysis of these bands showed amino acid sequences coinciding with those of phase-1 flagellin and hook-associated protein 2.

* Corresponding author. Mailing address: Department of Pharmacy, Uppsala University, P.O. Box 580, SE-751 23 Uppsala, Sweden. Phone: 46-18 471 4467. Fax: 46-18 471 4223. E-mail: ingvar.sjoholm{at}galenik.uu.se.

Infection and Immunity, March 2002, p. 1434-1442, Vol. 70, No. 3
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.3.1434-1442.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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