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JAC Advance Access originally published online on March 31, 2004
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Journal of Antimicrobial Chemotherapy (2004) 53, 863-866
© 2004 The British Society for Antimicrobial Chemotherapy

Effect of listeriolysin O-loaded erythrocytes on Mycobacterium avium replication within macrophages

L. Rossi1, G. Brandi2, M. Malatesta3, S. Serafini1, F. Pierigé1, A. G. Celeste2, G. F. Schiavano2, G. Gazzanelli3 and M. Magnani1,*

1 Institute of Biochemistry ‘G. Fornaini’, 2 Institute of Hygiene and 3 Institute of Histology and Laboratory Analyses, University of Urbino, Via Saffi, 2, 61029 Urbino (PU), Italy

Received 24 September 2003; returned 18 December 2003; revised 19 January 2004; accepted 4 February 2004

Objective: To evaluate the efficacy of erythrocytes loaded with the haemolytic toxin listeriolysin O against Mycobacterium avium replication within human macrophages.

Methods: Recombinant listeriolysin O was loaded in human erythrocytes by a procedure of hypotonic dialysis and isotonic resealing. Loaded erythrocytes were modified to allow them to be recognized and taken up by human macrophages infected with M. avium. The antimycobacterial activity of the erythrocytes loaded with listeriolysin O was evaluated by supernatant and intracellular cfu counts on days 4 and 7 post-erythrocyte administration.

Results: Recombinant listeriolysin O was encapsulated in human erythrocytes to reach final concentrations ranging from 1 to 4 ng/mL of erythrocytes. Erythrocytes loaded with increasing quantities of recombinant protein were able to reduce (at most by 50%) M. avium replication in a dose-dependent fashion when administered to infected macrophages.

Conclusions: Erythrocytes loaded with listeriolysin O are effective against M. avium replication within macrophages. We are confident that the strategy presented could be useful against mycobacteria other than M. avium (such as Mycobacterium tuberculosis and Mycobacterium leprae) by itself or as part of an antimycobacterial treatment.

Keywords: mycobacteria, mycobacterial phagosome, encapsulation, drug-targeting, phagocytosis

* Corresponding author. Tel: +39-722-305211; Fax: +39-722-320188; E-mail: magnani{at}uniurb.it


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