title: Heterogeneity of molecular resistance patterns in antimony-resistant field isolates of Leishmania species from the western Mediterranean area.
abstract: Antimonials remain the first-line treatment for the various manifestations of leishmaniasis in most areas where the disease is endemic, and increasing cases of therapeutic failure associated with parasite resistance have been reported. In this study, we assessed the molecular status of 47 clinical isolates of Leishmania causing visceral and cutaneous leishmaniasis from Algeria, Tunisia, and southern France. In total, we examined 14 genes that have been shown to exhibit significant variations in DNA amplification, mRNA levels, or protein expression with respect to resistance to antimonials. The gene status of each clinical isolate was assessed via qPCR and qRT-PCR. We then compared the molecular pattern against the phenotype determined via an in vitro sensitivity test of the clinical isolates against meglumine antimoniate, which is considered the reference technique. Our results demonstrate significant DNA amplification and/or RNA overexpression in 56% of the clinical isolates with the resistant phenotype. All clinical isolates that exhibited significant overexpression of at least 2 genes displayed a resistant phenotype. Among the 14 genes investigated, 10 genes displayed either significant amplification or overexpression in at least 1 clinical isolate; these genes are involved in several metabolic pathways. Moreover, various gene associations were observed depending on the clinical isolates, supporting the multifactorial nature of Leishmania resistance. Molecular resistance features were found in the 3 Leishmania species investigated (Leishmania infantum, Leishmania major, and Leishmania killicki). To our knowledge, this is the first report of the involvement of molecular resistance genes in field isolates of Leishmania major and Leishmania killicki with the resistance phenotype.
authors.affiliation: Laboratoire de Parasitologie-Mycologie, CHU Timone, UMR MD3 Aix-Marseille Université, Marseille, France fjeddi2@yahoo.fr.
authors.forename: Fakhri
authors.initials: F
authors.lastname: Jeddi
authors.affiliation: Laboratoire de Parasitologie-Mycologie, CHU Timone, UMR MD3 Aix-Marseille Université, Marseille, France.
authors.forename: Charles
authors.initials: C
authors.lastname: Mary
authors.affiliation: Laboratoire de recherche parasitoses médicales, Biotechnologie & Biomolécules, LR 11 IPT 06, Institut Pasteur de Tunis, Tunis, Tunisia.
authors.forename: Karim
authors.initials: K
authors.lastname: Aoun
authors.affiliation: Service Eco-épidémiologie Parasitaire et Génétique des Populations, Institut Pasteur d'Algérie, Algiers, Algeria Laboratoire Biodiversité et Environnement, Interactions, Génomes, USTHB, Algiers, Algeria.
authors.forename: Zoubir
authors.initials: Z
authors.lastname: Harrat
authors.affiliation: Laboratoire de recherche parasitoses médicales, Biotechnologie & Biomolécules, LR 11 IPT 06, Institut Pasteur de Tunis, Tunis, Tunisia.
authors.forename: Aïda
authors.initials: A
authors.lastname: Bouratbine
authors.affiliation: Laboratoire de Parasitologie-Mycologie, CHU Timone, UMR MD3 Aix-Marseille Université, Marseille, France.
authors.forename: Françoise
authors.initials: F
authors.lastname: Faraut
authors.affiliation: Service Eco-épidémiologie Parasitaire et Génétique des Populations, Institut Pasteur d'Algérie, Algiers, Algeria.
authors.forename: Rezika
authors.initials: R
authors.lastname: Benikhlef
authors.affiliation: INSERM, U1065, Centre Méditerranéen de Médecine Moléculaire, C3M, Toxines Microbiennes dans la relation hôte pathogènes, Nice, France Service de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Nice, Nice, France.
authors.forename: Christelle
authors.initials: C
authors.lastname: Pomares
authors.affiliation: Montpellier Université 1, UMR MIVEGEC (CNRS 5290/IRD 224/UM1/UM2), Montpellier, France CHRU de Montpellier, Montpellier, France.
authors.forename: Francine
authors.initials: F
authors.lastname: Pratlong
authors.affiliation: INSERM, U1065, Centre Méditerranéen de Médecine Moléculaire, C3M, Toxines Microbiennes dans la relation hôte pathogènes, Nice, France Service de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Nice, Nice, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Marty
authors.affiliation: Laboratoire de Parasitologie-Mycologie, CHU Timone, UMR MD3 Aix-Marseille Université, Marseille, France.
authors.forename: Renaud
authors.initials: R
authors.lastname: Piarroux
mesh_terms: D000462:Algeria; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D004351:Drug Resistance / Q000235:genetics*; D005602:France; D005786:Gene Expression Regulation; D005838:Genotype; D006801:Humans; D018314:Leishmania infantum / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D018320:Leishmania major / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D016773:Leishmaniasis, Cutaneous / Q000188:drug therapy / Q000469:parasitology; D007898:Leishmaniasis, Visceral / Q000188:drug therapy / Q000469:parasitology; D008536:Meglumine / Q000494:pharmacology*; D000077485:Meglumine Antimoniate; D053858:Metabolic Networks and Pathways / Q000235:genetics; D009942:Organometallic Compounds / Q000494:pharmacology*; D021261:Parasitic Sensitivity Tests; D010641:Phenotype; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D014416:Tunisia
chemical_list: D000981:Antiprotozoal Agents; D009942:Organometallic Compounds; D015800:Protozoan Proteins; D008536:Meglumine; D000077485:Meglumine Antimoniate