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Infection and Immunity, June 2000, p. 3129-3139, Vol. 68, No. 6
Department of Pathobiology, University of
Pennsylvania School of Veterinary Medicine, Philadelphia,
Pennsylvania 19104
Received 1 November 1999/Returned for modification 12 January
2000/Accepted 25 February 2000
Recombinant live oral vaccines expressing pathogen-derived antigens
offer a unique set of attractive properties. Among these are the
simplicity of administration, the capacity to induce mucosal and
systemic immunity, and the advantage of permitting genetic manipulation
for optimal antigen presentation. In this study, the benefit of having
a heterologous antigen expressed on the surface of a live vector rather
than intracellularly was evaluated. Accordingly, the immune response of
mice immunized with a Salmonella enterica serovar
Typhimurium vaccine strain expressing the Escherichia coli 987P fimbrial antigen on its surface (Fas+)
was compared with the expression in the periplasmic compartment (Fas
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Mucosal and Systemic Immune Responses to Chimeric Fimbriae
Expressed by Salmonella enterica Serovar Typhimurium
Vaccine Strains
). Orally immunized BALB/c mice showed that 987P
fimbriated Salmonella serovar Typhimurium CS3263
(aroA asd) with pCS151 (fas+
asd+) elicited a significantly higher level of
987P-specific systemic immunoglobulin G (IgG) and mucosal IgA than
serovar Typhimurium CS3263 with pCS152 (fasD mutant,
asd+) expressing 987P periplasmic antigen.
Further studies were aimed at determining whether the 987P fimbriae
expressed by serovar Typhimurium
4550 (cya crp asd)
could be used as carriers of foreign epitopes. For this, the vaccine
strain was genetically engineered to express chimeric fimbriae carrying
the transmissible gastroenteritis virus (TGEV) C (379-388) and A
(521-531) epitopes of the spike protein inserted into the 987P major
fimbrial subunit FasA. BALB/c mice administered orally serovar
Typhimurium
4550 expressing the chimeric fimbriae from the
tet promoter in pCS154 (fas+
asd+) produced systemic antibodies against both
fimbria and the TGEV C epitope but not against the TGEV A epitope. To
improve the immunogenicity of the chimeric fimbriae, the in vivo
inducible nirB promoter was inserted into pCS154, upstream
of the fas genes, to create pCS155. In comparison with the
previously used vaccine, BALB/c mice immunized orally with serovar
Typhimurium
4550/pCS155 demonstrated significantly higher levels of
serum IgG and mucosal IgA against 987P fimbria. Moreover, mucosal IgA
against the TGEV C epitope was only detected with serovar Typhimurium
4550/pCS155. The induced antibodies also recognized the epitopes in
the context of the full-length TGEV spike protein. Hence, immune
responses to heterologous chimeric fimbriae on
Salmonella vaccine vectors can be optimized by using
promoters known to be activated in vivo.
*
Corresponding author. Mailing address: University of
Pennsylvania School of Veterinary Medicine, 3800 Spruce St.,
Philadelphia, PA 19104-6049. Phone: (215) 898-1695. Fax: (215)
898-7887. E-mail: dmschiff{at}vet.upenn.edu.
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