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Entomopathogenic bacteria Photorhabdus luminescens as drug source against Leishmania amazonensis

  • Ana Maria Antonello (a1) (a2), Thaís Sartori (a1) (a2), Ana Paula Folmer Correa (a3), Adriano Brandelli (a3), Ralf Heermann (a4), Luiz Carlos Rodrigues Júnior (a2), Alessandra Peres (a2) (a5), Pedro Roosevelt Torres Romão (a2) (a6) (a7) and Onilda Santos Da Silva (a1)...

Abstract

Leishmaniasis is a widely spread and zoonotic disease with serious problems as low effectiveness of drugs, emergence of parasite resistance and severe adverse reactions. In recent years, considerable attention has been given to secondary metabolites produced by Photorhabdus luminescens, an entomopathogenic bacterium. Here, we assessed the leishmanicidal activity of P. luminescens culture fluids. Initially, promastigotes of Leishmania amazonensis were incubated with cell free conditioned medium of P. luminescens and parasite survival was monitored. Different pre-treatments of the conditioned medium revealed that the leishmanicidal activity is due to a secreted peptide smaller than 3 kDa. The Photorhabdus-derived leishmanicidal toxin (PLT) was enriched from conditioned medium and its effect on mitochondrial membrane potential of promastigotes, was determined. Moreover, the biological activity of PLT against amastigotes was evaluated. PLT inhibited the parasite growth and showed significant leishmanicidal activity against promastigote and amastigotes of L. amazonensis. PLT also caused mitochondrial dysfunction in parasites, but low toxicity to mammalian cell and human erythrocytes. Moreover, the anti-amastigote activity was independent of nitric oxide production. In summary, our results highlight that P. luminescens secretes Leishmania-toxic peptide(s) that are promising novel drugs for therapy against leishmaniasis.

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Corresponding author

Author for correspondence: Pedro Roosevelt Torres Romão and Onilda Santos da Silva, E-mail: pedror@ufcspa.edu.br and onilda.silva@ufrgs.br

References

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Bizani, D and Brandelli, A (2002) Characterization of a bacteriocin produced by a newly isolated Bacillus sp. Strain 8 A. Journal of Applied Microbiology 93, 512519.
Bode, E, Brachmann, AO, Kegler, C, Simsek, R, Dauth, C, Zhou, Q, Kaiser, M, Klemmt, P and Bode, HB (2015) Simple ‘on-demand’ production of bioactive natural products. Chembiochem 16, 11151119.
Bode, HB (2009) Entomopathogenic bacteria as a source of secondary metabolites. Current Opinion in Chemical Biology 13, 224230.
Brachmann, AO and Bode, HB (2013) Identification and bioanalysis of natural products from insect symbionts and pathogens. In Vilcinskas, A (ed.). Yellow Biotechnology I-Insect Biotechnologie in Drug Discovery and Preclinical Research, vol. 135. Berlin, Heidelberg: Springer, pp. 123155.
Bradford, MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248254.
Cai, X, Nowak, S, Wesche, F, Bischoff, I, Kaiser, M, Fürst, R and Bode, HB (2017) Entomopathogenic bacteria use multiple mechanisms for bioactive peptide library design. Nature Chemistry 9, 379386. http://www.nature.com/nchem/journal/v9/n4/abs/nchem.2671.html#supplementary-information.
Carvalho, EM, Barral, A, Costa, JML, Bittencourt, A and Marsden, P (1994) Clinical and immunopathological aspects of disseminated cutaneous leishmaniasis. Acta Tropica 56, 315325.
Castellani, A and Chalmers, AJ (1919) Manual of Tropical Medicine. 3rd edn. New York: WilIiam Wood, 2436 pp.
Challinor, VL and Bode, HB (2015) Bioactive natural products from novel microbial sources. Annals of the New York Academy of Sciences 1354, 8297.
Chappuis, F, Sundar, S, Hailu, A, Ghalib, H, Rijal, S, Peeling, RW, Alvar, J and Boelaert, M (2007) Visceral leishmaniasis: what are the needs for diagnosis, treatment and control? Nature Reviews Microbiology 5, 873882.
Chaston, JM, Suen, G, Tucker, SL, Andersen, AW, Bhasin, A, Bode, E, Bode, HB, Brachmann, AO, Cowles, CE, Cowles, KN, Darby, C, de Leon, L, Drace, K, Du, Z, Givaudan, A, Herbert Tran, EE, Jewell, KA, Knack, JJ, Krasomil-Osterfeld, KC, Kukor, R, Lanois, A, Latreille, P, Leimgruber, NK, Lipke, CM, Liu, R, Lu, X, Martens, EC, Marri, PR, Medigue, C, Menard, ML, Miller, NM, Morales-Soto, N, Norton, S, Ogier, JC, Orchard, SS, Park, D, Park, Y, Qurollo, BA, Sugar, DR, Richards, GR, Rouy, Z, Slominski, B, Slominski, K, Snyder, H, Tjaden, BC, van der Hoeven, R, Welch, RD, Wheeler, C, Xiang, B, Barbazuk, B, Gaudriault, S, Goodner, B, Slater, SC, Forst, S, Goldman, BS and Goodrich-Blair, H (2011) The entomopathogenic bacterial endosymbionts Xenorhabdus and Photorhabdus: convergent lifestyles from divergent genomes. PLoS ONE 6, e27909.
Cragg, GM and Newman, DJ (2013) Natural products: a continuing source of novel drug leads. Biochimica et Biophysica Acta 1830, 36703695.
Dagnino, AP, Barros, FMCd, Ccana-Ccapatinta, GV, Prophiro, JS, Poser, GLv and Romão, PRT (2015) Leishmanicidal activity of lipophilic extracts of some Hypericum species. Phytomedicine 22, 7176.
da Silva, OS, Prado, GR, da Silva, JL, Silva, CE, da Costa, M and Heermann, R (2013) Oral toxicity of Photorhabdus luminescens and Xenorhabdus nematophila (Enterobacteriaceae) against Aedes aegypti (Diptera: Culicidae). Parasitology Research 112, 28912896.
da Silva, JL, Undurraga Schwalm, F, Eugenio Silva, C, da Costa, M, Heermann, R and Santos da Silva, O (2017) Larvicidal and growth-inhibitory activity of entomopathogenic bacteria culture fluids against Aedes aegypti (Diptera: Culicidae). Journal of Economic Entomology 110, 378385.
Degrossoli, A, Arrais-Silva, WW, Colhone, MC, Gadelha, FR, Joazeiro, PP and Giorgio, S (2011) The influence of low oxygen on macrophage response to Leishmania infection. Scandinavian Journal of Immunology 74, 165175.
de Souza, W and Rodrigues, JC (2009) Sterol biosynthesis pathway as target for anti-trypanosomatid drugs. Interdisciplinary Perspectives on Infectious Diseases 2009, 642502.
Donia, MS, Ravel, J and Schmidt, EW (2008) A global assembly line for cyanobactins. Nature Chemical Biology 4, 341343.
El-Sadawy, HA, Forst, S, Abouelhag, HA, Ahmed, AM, Alajmi, RA and Ayaad, TH (2016) Molecular and phenotypic characterization of two bacteria, Photorhabdus luminescens subsp. akhurstii HRM1 and HS1 isolated from two entomopathogenic nematodes, Heterorhabditis indica RM1 and Heterorhabditis sp. S1. Pakistan Journal of Zoology 48, 5158.
Ferlini, C and Scambia, G (2007) Assay for apoptosis using the mitochondrial probes, Rhodamine123 and 10-N-nonyl acridine orange. Nature Protocols 2, 31113114.
Fischer-Le Saux, M, Viallard, V, Brunel, B, Normand, P and Boemare, NE (1999) Polyphasic classification of the genus Photorhabdus and proposal of new taxa: P. luminescens subsp. luminescens subsp. nov., P. luminescens subsp. akhurstii subsp. nov., P. luminescens subsp. laumondii subsp. nov., P. temperata sp. nov., P. temperata subsp. temperata subsp. nov. and P. asymbiotica sp. nov. International Journal of Systematic Microbiology 49(Pt 4), 16451656.
Fonseca, SG, Romao, PR, Figueiredo, F, Morais, RH, Lima, HC, Ferreira, SH and Cunha, FQ (2003) TNF-alpha mediates the induction of nitric oxide synthase in macrophages but not in neutrophils in experimental cutaneous leishmaniasis. European Journal of Immunology 33, 22972306.
Franca-Costa, J, Wanderley, JL, Deolindo, P, Zarattini, JB, Costa, J, Soong, L, Barcinski, MA, Barral, A and Borges, VM (2012) Exposure of phosphatidylserine on Leishmania amazonensis isolates is associated with diffuse cutaneous leishmaniasis and parasite infectivity. PLoS ONE 7, e36595.
Garcia, FP, Henrique da Silva Rodrigues, J, Din, ZU, Rodrigues-Filho, E, Ueda-Nakamura, T, Auzely-Velty, R and Nakamura, CV (2017) A3K2A3-induced apoptotic cell death of Leishmania amazonensis occurs through caspase- and ATP-dependent mitochondrial dysfunction. Apoptosis 22, 5771.
Gauthier, C, Legault, J, Girard-Lalancette, K, Mshvildadze, V and Pichette, A (2009) Haemolytic activity, cytotoxicity and membrane cell permeabilization of semi-synthetic and natural lupane- and oleanane-type saponins. Bioorganic & Medicinal Chemistry 17, 20022008.
Giorgio, S, Linares, E, de Capurro, ML, de Bianchi, AG and Augusto, O (1996) Formation of nitrosyl hemoglobin and nitrotyrosine during murine leishmaniasis. Photochemistry Photobiology 63, 750754.
Herbert, EE and Goodrich-Blair, H (2007) Friend and foe: the two faces of Xenorhabdus nematophila. Nature Reviews Microbiology 5, 634646.
Houghton, AM, Hartzell, WO, Robbins, CS, Gomis-Ruth, FX and Shapiro, SD (2009) Macrophage elastase kills bacteria within murine macrophages. Nature 460, 637641.
Kondo, S, Mizuki, E, Akao, T and Ohba, M (2002) Antitrichomonal strains of Bacillus thuringiensis. Parasitology Research 88, 10901092.
Kronenwerth, M, Brachmann, AO, Kaiser, M and Bode, HB (2014) Bioactive derivatives of isopropylstilbene from mutasynthesis and chemical synthesis. Chembiochem 15, 26892691.
Lee, N, Bertholet, S, Debrabant, A, Muller, J, Duncan, R and Nakhasi, HL (2002) Programmed cell death in the unicellular protozoan parasite Leishmania. Cell Death Differentiation 9, 5364.
Maruyama, C, Toyoda, J, Kato, Y, Izumikawa, M, Takagi, M, Shin-ya, K, Katano, H, Utagawa, T and Hamano, Y (2012) A stand-alone adenylation domain forms amide bonds in streptothricin biosynthesis. Nature Chemical Biology 8, 791797.
Nielsen-LeRoux, C, Gaudriault, S, Ramarao, N, Lereclus, D and Givaudan, A (2012) How the insect pathogen bacteria Bacillus thuringiensis and Xenorhabdus/Photorhabdus occupy their hosts. Current Opinion in Microbiology 15, 220231.
Novais, FO, Nguyen, BT, Beiting, DP, Carvalho, LP, Glennie, ND, Passos, S, Carvalho, EM and Scott, P (2014) Human classical monocytes control the intracellular stage of Leishmania braziliensis by reactive oxygen species. Journal of Infectious Diseases 209, 12881296.
Oh, S, Kim, S, Kong, S, Yang, G, Lee, N, Han, D, Goo, J, Siqueira-Neto, JL, Freitas-Junior, LH and Song, R (2014) Synthesis and biological evaluation of 2,3-dihydroimidazo[1,2-a]benzimidazole derivatives against Leishmania donovani and Trypanosoma cruzi. European Journal of Medicinal Chemistry 84, 395403.
Orozco, RA, Molnar, I, Bode, H and Stock, SP (2016) Bioprospecting for secondary metabolites in the entomopathogenic bacterium Photorhabdus luminescens subsp. sonorensis. Journal of Invertebrate Pathology 141, 4552.
Rodrigues, JHdS, Ueda-Nakamura, T, Corrêa, AG, Sangi, DP and Nakamura, CV (2014) A quinoxaline derivative as a potent chemotherapeutic agent, alone or in combination with benznidazole, against Trypanosoma cruzi. PLoS ONE 9, e85706.
Romao, PR, Fonseca, SG, Hothersall, JS, Noronha-Dutra, AA, Ferreira, SH and Cunha, FQ (1999) Glutathione protects macrophages and Leishmania major against nitric oxide-mediated cytotoxicity. Parasitology 118, 559566.
Romao, PR, Tovar, J, Fonseca, SG, Moraes, RH, Cruz, AK, Hothersall, JS, Noronha-Dutra, AA, Ferreira, SH and Cunha, FQ (2006) Glutathione and the redox control system trypanothione/trypanothione reductase are involved in the protection of Leishmania spp. against nitrosothiol-induced cytotoxicity. Brazilian Journal of Medical and Biological Research 39, 355363.
Sacks, D and Kamhawi, S (2001) Molecular aspects of parasite-vector and vector-host interactions in leishmaniasis. Annual Review of Microbiology 55, 453483.
Sen, R, Bandyopadhyay, S, Dutta, A, Mandal, G, Ganguly, S, Saha, P and Chatterjee, M (2007) Artemisinin triggers induction of cell-cycle arrest and apoptosis in Leishmania donovani promastigotes. Journal of Medical Microbiology 56, 12131218.
Shi, D, An, R, Zhang, W, Zhang, G and Yu, Z (2017) Stilbene derivatives from photorhabdus temperata SN259 and their antifungal activities against phytopathogenic fungi. Journal of Agricultural and Food Chemistry 65, 6065.
Shrestha, YK and Lee, KY (2012) Oral toxicity of Photorhabdus culture media on gene expression of the adult sweetpotato whitefly, Bemisia tabaci. Journal of Invertebrate Pathology 109, 9196.
Sieber, SA and Marahiel, MA (2005) Molecular mechanisms underlying nonribosomal peptide synthesis: approaches to new antibiotics. Chemical Reviews 105, 715738.
Sundar, S (2001) Drug resistance in Indian visceral leishmaniasis. Tropical Medicine & International Health 6, 849854.
Tobias, NJ, Mishra, B, Gupta, DK, Sharma, R, Thines, M, Stinear, TP and Bode, HB (2016) Genome comparisons provide insights into the role of secondary metabolites in the pathogenic phase of the Photorhabdus life cycle. BMC Genomics 17, 537.
Waterfield, NR, Ciche, T and Clarke, D (2009) Photorhabdus and a host of hosts. Annual Review of Microbiology 63, 557574.
Weiss, G and Schaible, UE (2015) Macrophage defense mechanisms against intracellular bacteria. Immunological Reviews 264, 182203.
World Health Organization (2010) Control of the Leishmaniasis: Report of a Meeting of the WHO Expert Committee on the Control of Leishmaniases. Geneva, Switzerland: World Health Organization.
Wu, G, Zhao, Z, Liu, C and Qiu, L (2014) Priming Galleria mellonella (Lepidoptera: Pyralidae) larvae with heat-killed bacterial cells induced an enhanced immune protection against Photorhabdus luminescens TT01 and the role of innate immunity in the process. Journal of Economic Entomology 107, 559569.
Xu, Z, Yao, B, Sun, M and Yu, Z (2004) Protection of mice infected with Plasmodium berghei by Bacillus thuringiensis crystal proteins. Parasitology Research 92, 5357.
Yamanaka, K, Maruyama, C, Takagi, H and Hamano, Y (2008) Epsilon-poly-L-lysine dispersity is controlled by a highly unusual nonribosomal peptide synthetase. Nature Chemical Biology 4, 766772.
Zhou, Q, Grundmann, F, Kaiser, M, Schiell, M, Gaudriault, S, Batzer, A, Kurz, M and Bode, HB (2013) Structure and biosynthesis of xenoamicins from entomopathogenic Xenorhabdus. Chemistry – A European Journal 19, 1677216779.

Keywords

Entomopathogenic bacteria Photorhabdus luminescens as drug source against Leishmania amazonensis

  • Ana Maria Antonello (a1) (a2), Thaís Sartori (a1) (a2), Ana Paula Folmer Correa (a3), Adriano Brandelli (a3), Ralf Heermann (a4), Luiz Carlos Rodrigues Júnior (a2), Alessandra Peres (a2) (a5), Pedro Roosevelt Torres Romão (a2) (a6) (a7) and Onilda Santos Da Silva (a1)...

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