Identification of antimony resistance markers in Leishmania tropica field isolates through a cDNA-AFLP approach.
Pentavalent antimonial compounds have been the first line therapy for leishmaniasis; unfortunately the rate of treatment failure of anthroponotic cutaneous leishmaniasis (ACL) is increasing due to emerging of drug resistance. Elucidation of the molecular mechanisms operating in antimony resistance is critical for development of new strategies for treatment. Here, we used a cDNA-AFLP approach to identify gene(s) which are differentially expressed in resistant and sensitive Leishmania tropica field isolates. We identified five genes, aquaglyceroporin (AQP1) acts in drug uptake, ATP-binding cassette (ABC) transporter (MRPA) involved in sequestration of drug, phosphoglycerate kinase (PGK) implicated in glycolysis metabolism, mitogen activated protein kinase (MAPK) and protein tyrosine phosphatase (PTP) responsible for phosphorylation pathway. The results were confirmed using real time RT-PCR which revealed an upregulation of MRPA, PTP and PGK genes and downregulation of AQP1 and MAPK genes in resistant isolate. To our knowledge, this is the first report of identification of PTP and PGK genes potentially implicated in resistance to antimonials. Our findings support the idea that distinct biomolecules might be involved in antimony resistance in L. tropica field isolates.
Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Elham
E
Kazemi-Rad
Mehdi
M
Mohebali
Mohammad Bagher
MB
Khadem-Erfan
Mojtaba
M
Saffari
Reza
R
Raoofian
Homa
H
Hajjaran
Ramtin
R
Hadighi
Ali
A
Khamesipour
Sassan
S
Rezaie
Hoda
H
Abedkhojasteh
Mansour
M
Heidari
D054458:Amplified Fragment Length Polymorphism Analysis; D000965:Antimony / Q000494:pharmacology; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D002460:Cell Line; D053938:DNA Fragmentation; D018076:DNA, Complementary / Q000737:chemistry / Q000378:metabolism; D016054:DNA, Protozoan / Q000737:chemistry / Q000378:metabolism; D004351:Drug Resistance / Q000235:genetics*; D017125:Genes, Protozoan; D020128:Inhibitory Concentration 50; D007895:Leishmania tropica / Q000187:drug effects* / Q000235:genetics; D008536:Meglumine / Q000494:pharmacology*; D000077485:Meglumine Antimoniate; D009942:Organometallic Compounds / Q000494:pharmacology*; D021261:Parasitic Sensitivity Tests; D016053:RNA, Protozoan / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction
D000981:Antiprotozoal Agents; D018076:DNA, Complementary; D016054:DNA, Protozoan; D009942:Organometallic Compounds; D016053:RNA, Protozoan; D008536:Meglumine; D000077485:Meglumine Antimoniate; D000965:Antimony
Antimony resistance; Leishmania tropica; Real-time RT-PCR; cDNA-AFLP