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Infection and Immunity, October 2001, p. 6296-6302, Vol. 69, No. 10
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.10.6296-6302.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Reassessing the Role of Staphylococcus aureus Clumping Factor and Fibronectin-Binding Protein by Expression in Lactococcus lactis

Yok-Ai Que,1 Patrice François,2 Jacques-Antoine Haefliger,3 José-Manuel Entenza,1 Pierre Vaudaux,2 and Philippe Moreillon1,*

Division of Infectious Diseases1 and Laboratory of Molecular Biology,3 Department of Internal Medicine, Centre Hospitalier Universitaire Vaudois, 1011 Lausanne, and Division of Infectious Diseases, University Hospital of Geneva, 1211 Geneva 14,2 Switzerland

Received 2 April 2001/Returned for modification 1 May 2001/Accepted 26 June 2001

Since Staphylococcus aureus expresses multiple pathogenic factors, studying their individual roles in single-gene-knockout mutants is difficult. To circumvent this problem, S. aureus clumping factor A (clfA) and fibronectin-binding protein A (fnbA) genes were constitutively expressed in poorly pathogenic Lactococcus lactis using the recently described pOri23 vector. The recombinant organisms were tested in vitro for their adherence to immobilized fibrinogen and fibronectin and in vivo for their ability to infect rats with catheter-induced aortic vegetations. In vitro, both clfA and fnbA increased the adherence of lactococci to their specific ligands to a similar extent as the S. aureus gene donor. In vivo, the minimum inoculum size producing endocarditis in >= 80% of the rats (80% infective dose [ID80]) with the parent lactococcus was >= 107 CFU. In contrast, clfA-expressing and fnbA-expressing lactococci required only 105 CFU to infect the majority of the animals (P < 0.00005). This was comparable to the infectivities of classical endocarditis pathogens such as S. aureus and streptococci (ID80 = 104 to 105 CFU) in this model. The results confirmed the role of clfA in endovascular infection, but with a much higher degree of confidence than with single-gene-inactivated staphylococci. Moreover, they identified fnbA as a critical virulence factor of equivalent importance. This was in contrast to previous studies that produced controversial results regarding this very determinant. Taken together, the present observations suggest that if antiadhesin therapy were to be developed, at least both of the clfA and fnbA products should be blocked for the therapy to be effective.

* Corresponding author. Mailing address: Division of Infectious Diseases, Department of Internal Medicine, Centre Hospitalier Universitaire Vaudois, 1011 Lausanne, Switzerland. Phone: 41-21-314-10-26. Fax: 41-21-314-10-36. E-mail: pmoreill{at}chuv.hospvd.ch.

Infection and Immunity, October 2001, p. 6296-6302, Vol. 69, No. 10
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.10.6296-6302.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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