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Cross-resistance of Leishmania infantum isolates to nitric oxide from patients refractory to antimony treatment, and greater tolerance to antileishmanial responses by macrophages

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Abstract

Visceral leishmaniasis is a life-threatening disease characterized by intense parasitism of the spleen, liver, and bone marrow. Antimonials have served as front-line antileishmanial therapeutics for decades, but the increasing failure rates under antimonial treatment have challenged the continued use of these drugs. Pentavalent antimonials are known to reinforce the killing mechanisms of macrophages, although the associated mechanism remains unclear. Here, for the first time, we determined whether Leishmania infantum strains isolated from patients refractory to antimony treatment (relapse cases) were cross-resistant to antimonials, liposomal amphotericin B, and/or nitric oxide, and also whether these strains modulate macrophage infection. We selected four clinical isolates from relapse cases and two clinical isolates from antimony-responsive patients (control group) for the present study. The L. infantum promastigotes from all four relapse cases were resistant to trivalent antimonial treatment and nitric oxide, while only one isolate was resistant to liposomal amphotericin B. We evaluated whether the resistant strains from relapse cases showed enhanced infectivity and amastigote survival in macrophages, or macrophage-killing mechanisms in macrophages activated by lipopolysaccharide plus interferon gamma. Infection indexes calculated using macrophages infected with isolates from relapse were higher than those observed with control strains that were stimulated independently. Macrophage infection was higher with L. infantum isolates from relapse cases and correlated with enhanced interleukin 1-β production but showed similar nitrite production. Our results demonstrate that L. infantum field isolates from relapse cases were resistant to antimonials and nitric oxide and that these parasites stimulated inflammatory cytokines and were resistant to macrophage-killing mechanisms, factors that may contribute to disease severity.

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Acknowledgments

We thank members of the Infectious Disease Clinic Group from the University Hospital, Universidade Federal de Sergipe.

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Authors and Affiliations

  1. Laboratório de Biologia Molecular, Hospital Universitário, Universidade Federal de Sergipe, Aracaju, 49060-10, Brazil

    Tatiana R. de Moura, Micheli Luize Barbosa Santos, Juciene M. Braz, Luis Felipe V. C. Santos, Matheus T. Aragão, Fabricia A. de Oliveira, Priscila L. Santos, Ângela Maria da Silva, Amélia Ribeiro de Jesus & Roque P. de Almeida

  2. Instituto de Investigação em Imunologia, Sao Paulo, 01246-903, Brazil

    Amélia Ribeiro de Jesus & Roque P. de Almeida

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  1. Tatiana R. de Moura
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  2. Micheli Luize Barbosa Santos
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  3. Juciene M. Braz
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  4. Luis Felipe V. C. Santos
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  9. Amélia Ribeiro de Jesus
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  10. Roque P. de Almeida
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Correspondence to Tatiana R. de Moura.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Ethical approval was obtained from the Hospital Universitário from Universidade Federal de Sergipe, CAAE 0151.0.107.000-07.

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de Moura, T.R., Santos, M.L.B., Braz, J.M. et al. Cross-resistance of Leishmania infantum isolates to nitric oxide from patients refractory to antimony treatment, and greater tolerance to antileishmanial responses by macrophages. Parasitol Res 115, 713–721 (2016). https://doi.org/10.1007/s00436-015-4793-4

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