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Infection and Immunity, July 2002, p. 3576-3585, Vol. 70, No. 7
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.7.3576-3585.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification of a Disulfide Isomerase Protein of Leishmania major as a Putative Virulence Factor

Y. Ben Achour, M. Chenik, H. Louzir,* and K. Dellagi

Laboratoire d'Immunologie (LAF301), Institut Pasteur de Tunis, 1002 Tunis-Belvédère, Tunisia

Received 26 December 2001/ Returned for modification 25 February 2002/ Accepted 29 March 2002

Several approaches have been previously used to elucidate the genetic basis of Leishmania virulence. In general, they were based on laboratory Leishmania clones genetically modified or grown in the presence of selecting agents. In a previous study, we demonstrated that Leishmania major freshly isolated from human cutaneous lesions showed significant differences in the severity of the experimental disease induced in BALB/c mice. Here, using the mRNA differential display technique, we analyzed gene expression in L. major promastigotes showing different levels of virulence. We have identified a novel Leishmania gene encoding a 477-amino-acid protein exhibiting two distinct regions that are identical to the putative active-site sequence (CGHC) of the eukaryotic protein disulfide isomerase (PDI). The recombinant protein displayed a specific PDI enzymatic activity. This L. major disulfide isomerase protein (LmPDI) is predominantly expressed, at both the mRNA and protein levels, in highly virulent strains. Specific PDI inhibitors abolished the enzymatic activity of the recombinant protein and profoundly affected parasite growth. These findings suggest that LmPDI may play an important role in Leishmania natural pathogenicity and may constitute a new target for anti-Leishmania chemotherapy.

* Corresponding author. Mailing address: Laboratoire d'Immunologie, Institut Pasteur de Tunis, BP: 74, 1002 Tunis-Belvédère, Tunisia. Phone: 216-1 783022. Fax: 216-1 791833. E-mail: hechmi.louzir{at}pasteur.rns.tn.

Editor: W. A. Petri, Jr.

Infection and Immunity, July 2002, p. 3576-3585, Vol. 70, No. 7
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.7.3576-3585.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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