Expression Analysis of Multiple Genes May Involve in Antimony Resistance among Leishmania major Clinical Isolates from Fars Province, Central Iran

  • Nafiseh GHOBAKHLOO Dept. of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
  • Mohammad Hossein MOTAZEDIAN Mail Basic Sciences in Infectious Diseases, Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
  • Majid FARDAEI Dept. of Genetics, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Antimony resistance, Leishmania major, Molecular marker, Iran


Background: Treatment of Cutaneous Leishmaniasis (CL) is being faced with serious difficulties in Fars Province, due to emerging of resistance against meglumine antimonite (Glucantime®). In this context, determining some biomarkers for drug sensitivity monitoring seems to be highly essential. Different studies have been carried out to decipher the genes might be involved in antimony resistant phenotype in Leishmania spp. Here, we selected three genes: AQP (as drug transporter), TDR-1-1(as drug activator), and γ-GCS (inducing reduction environment) for comparative expression analysis on clinical resistant and sensitive isolates of L. major.

Methods: The clinical isolates of L. major were collected from CL patients referred to Valfajr Health Center, Shiraz from Oct 2011 to Feb 2012. The susceptibility test was performed to confirm drug sensitivity of strains in vitro as well. Then, the gene expression analysis was performed by quantitative real-time PCR using SYBR® Green.

Results: By comparison of expression level between strains, up regulation of γ-GCS gene and down regulation of AQP gene were observed in resistant strains compared to the sensitive isolates; however, down regulation of AQP was not statistically specific. Analysis of TDR-1-1 gene unexpectedly showed a high level of expression in the non-responsive cases.

Conclusion: The γ-GCS, at least, can be considered as a suitable molecular marker for screening antimony sensitivity in clinical isolates, although AQP and TDR-1-1gene seem not to be reliable resistant markers. 


Alvar J, Velez ID, Bern C, Herrero M, Desjeux P, Cano J, et al. Leishmaniasis worldwide and global estimates of its incidence. PloS One. 2012; 7(5):e35671.

Akhoundi M, Hajjaran H, Baghaei A, Mohebali M. Geographical distribution of Leishmania species of human cutaneous leishmaniasis in Fars Province, southern Iran. Iran J Parasitol. 2013; 8(1): 85-91.

Sereno D, Maia C, Aït-Oudhia K. Antimony resistance and environment: Elusive links to explore during Leishmania life cycle. Int J Parasitol Drugs Drug Resist. 2012; 2: 200-203.

Masmoudi A, Hariz W, Marrekchi S, Amouri M, Turki H. Old world cutaneous leishmaniasis: diagnosis and treatment. J Dermatol Case Rep. 2013; 7(2):31-41.

Hadighi R, Boucher P, Khamesipour A, Meamar A, Roy G, Ouellette M, et al. Glucantime-resistant Leishmania tropica isolated from Iranian patients with cutaneous leishmaniasis are sensitive to alternative antileishmania drugs. Parasitol Res. 2007; 101(5):1319-1322.

Mohammadzadeh M, Behnaz F, Golshan Z. Efficacy of glucantime for treatment of cutaneous leishmaniasis in Central Iran. J Infect Public Health. 2013; 6(2):120-124.

Pourmohammadi B, Motazedian M, Handjani F, Hatam G, Habibi S, Sarkari B. Glucantime efficacy in the treatment of zoonotic cutaneous leishmaniasis. Southeast Asian J Trop Med Public Health. 2011; 42(3):502-508.

Sundar S, Singh A, Singh OP. Strategies to overcome Antileishmanial Drugs unresponsiveness. J Trop Med. 2014; 2014:646932.

Decuypere S, Vanaerschot M, Brunker K, Imamura H, Müller S, Khanal B, et al. Molecular mechanisms of drug resistance in natural Leishmania populations vary with genetic background. PLoS Negl Trop Dis. 2012; 6(2):e1514.

Aït-Oudhia K, Gazanion E, Vergnes B, Oury B, Sereno D. Leishmania antimony resistance: what we know what we can learn from the field. Parasitol Res. 2011; 109(5):1225-1232.

Jeddi F, Piarroux R, Mary C. Antimony resistance in Leishmania, focusing on experi-mental research. J Trop Med. 2011; 2011:695382.

Gourbal B, Sonuc N, Bhattacharjee H, Legare D, Sundar S, Ouellette M, et al. Drug uptake and modulation of drug resistance in Leishmania by an aquaglyceroporin. J Biol Chem. 2004; 279(30):31010-7.

Denton H, McGregorJ, COOMBS G. Reduction of anti-leishmanial pentavalent antimonial drugs by a parasite-specific thiol-dependent reductase, TDR-1. Biochem J. 2004; 381:405-12.

Ashutosh, Sundar S, Goyal N. Molecular mechanisms of antimony resistance in Leishmania. J Med Microbiol. 2007; 56(2):143-153.

Kazemi-Rad E, Mohebali M, Khadem-Erfan MB, Saffari M, Raoofian R, Hajjaran H, et al. Identification of antimony resistance markers in Leishmania tropica field isolates through a cDNA-AFLP approach. Exp Parasitol. 2013; 135(2):344-349.

Kulshrestha A, Singh R, Kumar D, Negi NS, Salotra P. Antimony-resistant clinical isolates of Leishmania donovani are susceptible to paromomycin and sitamaquine. Antimicrob Agents Chemother. 2011; 55(6):2916-2921.

Da Luz RI, Vermeersch M, Dujardin J-C, Cos P, Maes L. In vitro sensitivity testing of Leishmania clinical field isolates: preconditioning of promastigotes enhances infectivity for macrophage host cells. Antimicrob Agents Chemother. 2009; 53(12):5197-203.

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method.Methods. 2001, 25(4):402-408.

Askarian M, Mansour Ghanaie R, Karimi A, Habibzadeh F. Infectious diseases in Iran: a bird’s eye view. Clin Microbiol Infect. 2012; 18(11):1081-1088.

Mukhopadhyay R, Mandal G, Atluri VSR, Figarella K, Uzcategui NL, Zhou Y, et al. The role of alanine 163 in solute permeability of Leishmania major aquaglyceroporin LmAQP1. Mol Biochem Parasitol. 2011; 175(1):83-90.

Maharjan M, Singh S, Chatterjee M, Madhubala R. Role of aquaglyceroporin (AQP1) gene and drug uptake in antimony-resistant clinical isolates of Leishmania donovani. Am J Trop Med Hyg. 2008; 79(1):69-75.

Marquis N, Gourbal B, Rosen BP, Mukho-padhyay R, Ouellette M. Modulation in aquaglyceroporin AQP1 gene transcript levels in drug‐resistant Leishmania. Mol Microbiol. 2005; 57(6):1690-1699.

Adaui V, Schnorbusch K, Zimic M, Gutiérrez A, Decuypere S, Vanaerschot M, et al. Comparison of gene expression patterns among Leishmania braziliensis clinical isolates showing a different in vitro susceptibility to pentavalent antimony. Parasitology.2011; 138(02):183-193.

Mittal MK, Rai S, Gupta S et al. Characterization of natural antimony resistance in Leishmania donovani isolates. Am J Trop Med Hyg. 2007; 76(4):681-688.

Wyllie S, Mandal G, Singh N, Sundar S, Fairlamb AH, Chatterjee M. Elevated levels of tryparedoxin peroxidase in antimony unresponsive Leishmania donovani field isolates. Mol Biochem Parasitol. 2010; 173(2):162-164.

Decuypere S, Rijal S, Yardley V, De Doncker S, Laurent T, Khanal B, et al. Gene expression analysis of the mechanism of natural Sb (V) resistance in Leishmania donovani isolates from Nepal. Antimicrob Agents Chemother. 2005;49(11):4616-4621.

Mukherjee A, Padmanabhan PK, Singh S, Roy G, Girard I, Chatterjee M, et al. Role of ABC transporter MRPA, γ-glutamyl cysteine synthetase and ornithine decarboxylase in natural antimony-resistant isolates of Leishmania donovani. J Antimicrob Chemother. 2007; 59(2):204-211.

Torres DC, Adaui V, Ribeiro-Alves M, Romero GA, Arévalo J, Cupolillo E, et al. Targeted gene expression profiling in Leishmani braziliensis and Leishmania guyanensis parasites isolated from Brazilian patients with different antimonial treatment outcomes. Infect Genet Evol. 2010; 10(6):727-733.

Kumar D, Singh R, Bhandari V, Kulshrestha A, Negi NS, Salotra P. Biomarkers of antimony resistance: need for expression analysis of multiple genes to distinguish resistance phenotype in clinical isolates of Leishmania donovani. Parasitol Res. 2012; 111(1):223-230.

Andrews KT, Fisher G, Skinner-Adams TS. Drug repurposing and human parasitic protozoan diseases. Int J Parasitol Drugs Drug Resist. 2014; 4(2):95-111.

How to Cite
GHOBAKHLOO N, MOTAZEDIAN MH, FARDAEI M. Expression Analysis of Multiple Genes May Involve in Antimony Resistance among Leishmania major Clinical Isolates from Fars Province, Central Iran. Iran J Parasitol. 11(2):168-176.
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