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Effect of bovine lactoferricin on enteropathogenic Yersinia adhesion and invasion in HEp-2 cells

¹Laboratory of Ultrastructure, Istituto Superiore di Sanità, Viale Regina Elena, 299, 00161 Rome, Italy ²Department of Biology, III University of Rome, Italy ³Department of Experimental Medicine, II University of Naples, Naples, Italy ⁴Department of Public Health Sciences, University ‘La Sapienza', Rome, Italy

Correspondence Fabiana Superti superti{at}iss.it

Received August 1, 2003
Accepted January 23, 2004

Bovine lactoferricin, a pepsin-generated antimicrobial peptide from bovine lactoferrin active against a wide range of bacteria, was tested for its ability to influence the adhesion and invasion of Yersinia enterocolitica and Yersinia pseudotuberculosis in HEp-2 cells. The addition of non-cytotoxic and non-bactericidal concentrations of lactoferricin to cell monolayers before infection, under different bacterial growth experimental conditions, was ineffective or resulted in about a 10-fold increase in bacterial adhesion, whereas, in bacteria grown in conditions allowing maximal inv gene expression, a 10-fold inhibition of cell invasion by lactoferricin was observed. To confirm that the anti-invasive activity of lactoferricin was exerted against invasin-mediated bacterial entry, experiments were also performed utilizing Escherichia coli strain HB101 (pRI203), harbouring the inv gene from Y. pseudotuberculosis, which allows penetration of mammalian cells. Under these experimental conditions, lactoferricin was able to inhibit bacterial entry into epithelial cells, demonstrating that this peptide acts on inv-mediated Yersinia species invasion. As the inv gene product is the most important virulence factor in enteropathogenic Yersinia, being responsible for bacterial adherence and penetration within epithelial cells of the intestinal lumen and for the subsequent colonization of regional lymph nodes, these data provide additional information on the protective role of lactoferricin against bacterial infection.

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