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JAC Advance Access published online on March 31, 2004
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkh164
© 2004 by The British Society for Antimicrobial Chemotherapy
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© 2004 The British Society for Antimicrobial Chemotherapy

Brief report

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

L. Rossi 1 , G. Brandi 2 , M. Malatesta 3 , S. Serafini 1 , F. Pierigé 1 , A. G. Celeste 2 , G. F. Schiavano 2 , G. Gazzanelli 3 , and M. Magnani 1 *

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

* Corresponding author. E-mail: magnani{at}uniurb.it.

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

Abstract

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
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