• 1.

    Alvar J, Velez ID, Herrero M, Desjeux P, Cano J, Janin J, den Boer M; WHO Leishmaniasis Control Team, 2012. Leishmaniasis worldwide and global estimates of its incidence. PLoS ONE 7: e35671.

    • Search Google Scholar
    • Export Citation
  • 2.

    Guerin PJ, Olliaro P, Sundar S, Boelaert M, Croft SL, Desjeux P, Wasunna MK, Bryceson AD, 2002. Visceral leishmaniasis: current status of control, diagnosis, and treatment, and a proposed research and development agenda. Lancet Infect Dis 2: 494501.

    • Search Google Scholar
    • Export Citation
  • 3.

    Joshi A, Narain JP, Prasittisuk C, Bhatia R, Hashim G, Jorge A, Banjara M, Kroeger A, 2008. Can visceral leishmaniasis be eliminated from Asia? J Vector Borne Dis 45: 105111.

    • Search Google Scholar
    • Export Citation
  • 4.

    WHO, 2005. Expert Committee on Regional Strategic Framework for Elimination of Kala-azar from the South-East Asia Region (2005–2015). New Delhi: WHO regional Office for South-East Asia.

    • Search Google Scholar
    • Export Citation
  • 5.

    Prajapati VK, Awasthi K, Gautam S, Yadav TP, Rai M, Srivastava ON, Sundar S, 2011. Targeted killing of Leishmania donovani in vivo and in vitro with amphotericin B attached to functionalized carbon nanotubes. J Antimicrob Chemother 66: 874879.

    • Search Google Scholar
    • Export Citation
  • 6.

    Sundar S, Jha TK, Thakur CP, Engel J, Sindermann H, Fischer C, Junge K, Bryceson A, Berman J, 2002. Oral miltefosine for Indian visceral leishmaniasis. N Engl J Med 347: 17391746.

    • Search Google Scholar
    • Export Citation
  • 7.

    Jha TK, Sundar S, Thakur CP, Bachmann P, Karbwang J, Fischer C, Voss A, Berman J, 1999. Miltefosine, an oral agent, for the treatment of Indian visceral leishmaniasis. N Engl J Med 341: 17951800.

    • Search Google Scholar
    • Export Citation
  • 8.

    Sundar S, Singh A, Rai M, Prajapati VK, Singh AK, Ostyn B, Boelaert M, Dujardin JC, Chakravarty J, 2012. Efficacy of miltefosine in the treatment of visceral leishmaniasis in India after a decade of use. Clin Infect Dis 55: 543550.

    • Search Google Scholar
    • Export Citation
  • 9.

    Rijal S, Ostyn B, Uranw S, Rai K, Bhattarai NR, Dorlo TP, Beijnen JH, Vanaerschot M, Decuypere S, Dhakal SS, Das ML, Karki P, Singh R, Boelaert M, Dujardin JC, 2013. Increasing failure of Miltefosine in the treatment of kala-azar in Nepal and the potential role of parasite drug resistance, reinfection, or noncompliance. Clin Infect Dis 56: 15301538.

    • Search Google Scholar
    • Export Citation
  • 10.

    Kumar D, Kulshrestha A, Singh R, Salotra P, 2009. In vitro susceptibility of field isolates of Leishmania donovani to Miltefosine and amphotericin B: correlation with sodium antimony gluconate susceptibility and implications for treatment in areas of endemicity. Antimicrob Agents Chemother 53: 835838.

    • Search Google Scholar
    • Export Citation
  • 11.

    Prajapati VK, Mehrotra S, Gautam S, Rai M, Sundar S, 2012. In vitro antileishmanial drug susceptibility of clinical isolates from patients with Indian visceral leishmaniasis–status of newly introduced drugs. Am J Trop Med Hyg 87: 655657.

    • Search Google Scholar
    • Export Citation
  • 12.

    Kulshrestha A, Bhandari V, Mukhopadhyay R, Ramesh V, Sundar S, Maes L, Dujardin JC, Roy S, Salotra P, 2013. Validation of a simple resazurin-based promastigote assay for the routine monitoring of miltefosine susceptibility in clinical isolates of Leishmania donovani. Parasitol Res 112: 825828.

    • Search Google Scholar
    • Export Citation
  • 13.

    Maurya R, Mehrotra S, Prajapati VK, Nylen S, Sacks D, Sundar S, 2010. Evaluation of blood agar microtiter plates for culturing Leishmania parasites to titrate parasite burden in spleen and peripheral blood of patients with visceral leishmaniasis. J Clin Microbiol 48: 19321934.

    • Search Google Scholar
    • Export Citation
  • 14.

    Srivastava P, Prajapati VK, Vanaerschot M, Van der Auwera G, Dujardin JC, Sundar S, 2010. Detection of Leptomonas sp. parasites in clinical isolates of Kala-azar patients from India. Infect Genet Evol 10: 11451150.

    • Search Google Scholar
    • Export Citation
  • 15.

    Bhandari V, Kulshrestha A, Deep DK, Stark O, Prajapati VK, Ramesh V, Sundar S, Schonian G, Dujardin JC, Salotra P, 2012. Drug susceptibility in Leishmania isolates following miltefosine treatment in cases of visceral leishmaniasis and post kala-azar dermal leishmaniasis. PLoS Negl Trop Dis 6: e1657.

    • Search Google Scholar
    • Export Citation
  • 16.

    Leprohon P, Legare D, Raymond F, Madore E, Hardiman G, Corbeil J, Ouellette M, 2009. Gene expression modulation is associated with gene amplification, supernumerary chromosomes and chromosome loss in antimony-resistant Leishmania infantum. Nucleic Acids Res 37: 13871399.

    • Search Google Scholar
    • Export Citation
 
 
 
 

 

 
 

 

 

 

 

 

 

In vitro Susceptibility of Leishmania donovani to Miltefosine in Indian Visceral Leishmaniasis

View More View Less
  • Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221 005, UP, India; Unit of Epidemiology and Control of Tropical Diseases, Department of Public Health, Institute of Tropical Medicine, Antwerp, Belgium; National Institute of Pathology, Indian Council of Medical Research, Safdarjung Hospital Campus, New Delhi, India; Unit of Molecular Parasitology, Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium; Department of Biomedical Sciences, Antwerp University, Antwerp, Belgium

Promastigote miltefosine (MIL) susceptibility was performed on Leishmania donovani isolates from Indian patients with visceral leishmaniasis treated with MIL. Isolates that were obtained before the onset of MIL treatment, after completion of treatment (29th day), or at the time of treatment failure, were screened using in vitro promastigote assay. The MIL susceptibility of the pre-treatment isolates (N = 24, mean IC50 ± SEM = 3.74 ± 0.38 μM) was significantly higher than that of the post-treatment group (N = 26, mean IC50 ± SEM = 6.15 ± 0.52 μM; P = 0.0006) but was similar in the cured patients (N = 22, mean IC50 ± SEM = 5.58 ± 0.56 μM) and those who failed treatment (N = 28, mean IC50 ± SEM = 4.53 ± 0.47 μM). The pre/post-treatment results thus showed a 2-fold difference, whereas isolated from cured versus failed patients showed a similar susceptibility, suggesting that this higher tolerance is not responsible for MIL-treatment failure. Our work highlights the need for careful monitoring of MIL susceptibility for implementation in national VL elimination programs.

Author Notes

* Address correspondence to Shyam Sundar, Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221 005, UP, India. E-mail: drshyamsundar@hotmail.com

Financial support: This work was funded by Kaladrug-R (grant no. EC-FP7-222895, www.leishrisk.net/kaladrug), Directorate-General for Development Cooperation of the Belgian Government (framework agreement 03 - project 3.04 Visceral Leishmaniasis Control), Sitaram Memorial Trust, Muzaffarpur. Author V.K.P. is thankful to Indian Council of Medical Research, New Delhi, India for providing research fellowship. S.S. has received grant support for clinical trials and travel funds to attend scientific meetings from Paladin Labs, Institute for One World Health, and GlaxoSmithKline, and his institute has received grants from Bharat Serum and Vaccine Ltd.

Authors' addresses: Vijay Kumar Prajapati, Mitchell Cancer Institute, University of South Alabama, Mobile, AL, E-mail: vijay84bhu@gmail.com. Bart Ostyn, Unit of Epidemiology and Control of Tropical Diseases, Department of Public Health, Institute of Tropical Medicine, Antwerp, Belgium, E-mail: Bostyn@itg.be. Poonam Salotra, National Institute of Pathology, Indian Council of Medical Research, Safdarjung Hospital Campus, New Delhi, India, E-mail: salotra@vsnl.com. Manu Vanaerschot, Unit of Molecular Parasitology, Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium, E-mail: Mvanaerschot@itg.be. Jean-Claude Dujardin, Unit of Molecular Parasitology, Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium, and the Department of Biomedical Sciences, Antwerp University, Antwerp, Belgium, E-mail: Jcdujardin@itg.be. Smriti Sharma, Madhukar Rai, and Shyam Sundar, Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, UP, India, E-mails: smritibhardwaj87@gmail.com, rai_madhukar@sify.com, and drshyamsundar@hotmail.com.

Save