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Infection and Immunity, June 2000, p. 3251-3260, Vol. 68, No. 6
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Heterologous Expression of an Immunogenic
Pneumococcal Type 3 Capsular Polysaccharide in Lactococcus
lactis
Christophe
Gilbert,*
Karen
Robinson,
Richard W. F.
Le
Page, and
Jeremy M.
Wells
Cortecs Centre for Vaccine Discovery,
Department of Pathology, University of Cambridge, Cambridge CB2
1QP, United Kingdom
Received 21 June 1999/Returned for modification 27 July
1999/Accepted 23 March 2000
In order to develop a new system for the analysis of capsular
biosynthetic pathways we have explored the possibility of expressing type 3 capsular polysaccharide (CPS) from the pathogen
Streptococcus pneumoniae in Lactococcus lactis,
an unencapsulated lactic acid bacterium being developed as a vaccine
delivery vehicle for mucosal immunization. Only three of the four type
3 CPS biosynthesis genes were found to be necessary for the abundant
formation (120 mg liter
1) of an extracellular type 3 CPS
in L. lactis, implying a role for the type 3-specific
synthase in the extracellular transport of the CPS or implying the
existence of an alternative export system in L. lactis. The
authenticity of the expressed heterologous polysaccharide was
established by chemical and immunological analyses. Proton and carbon
nuclear magnetic resonance spectroscopy of CPSs purified from L. lactis and S. pneumoniae showed that the two CPS
structures were identical. When mice were immunized intraperitoneally with 3.5 × 106 CFU of live recombinant lactococci
expressing a total of approximately 0.5 µg of type 3 CPS, the immune
responses elicited appeared identical to those observed in mice
inoculated with 0.5 µg of type 3 CPS purified from S. pneumoniae. These findings show that L. lactis is a
useful host in which to study the role and function of genes involved
in the production of bacterial capsules. Additionally, L. lactis shows potential as a host for the safe production of capsule antigens and as a vaccine delivery vehicle for polysaccharide antigens.
*
Corresponding author. Mailing address: Laboratoire de
Microbiologie et Génétique Moléculaire, Bât.
405, 3ème étage, Université Lyon 1, 43, boulevard du
11 novembre 1918, 69622 Villeurbanne cedex, France. Phone: (33) 4 72 43 13 66. Fax: (33) 4 72 43 26 86. E-mail:
gilbert{at}biomserv.univ-lyon1.fr.
Infection and Immunity, June 2000, p. 3251-3260, Vol. 68, No. 6
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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