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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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