Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
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A Novel Anti-Microbial Peptide from Pseudomonas, REDLK Induced Growth Inhibition of Leishmania tarentolae Promastigote in Vitro

  • Yu, Yanhui (Key Laboratory of Zoonosis Research by Ministry of Education, College of Veterinary Medicine, Jilin University) ;
  • Zhao, Panpan (Key Laboratory of Zoonosis Research by Ministry of Education, College of Veterinary Medicine, Jilin University) ;
  • Cao, Lili (Key Laboratory of Zoonosis Research by Ministry of Education, College of Veterinary Medicine, Jilin University) ;
  • Gong, Pengtao (Key Laboratory of Zoonosis Research by Ministry of Education, College of Veterinary Medicine, Jilin University) ;
  • Yuan, Shuxian (Jilin Academy of Animal Husbandry and Veterinary Medicine) ;
  • Yao, Xinhua (Jilin Academy of Animal Husbandry and Veterinary Medicine) ;
  • Guo, Yanbing (Jilin Academy of Animal Husbandry and Veterinary Medicine) ;
  • Dong, Hang (Jilin Academy of Animal Husbandry and Veterinary Medicine) ;
  • Jiang, Weina (Department of Pathology, Qingdao Municipal Hospital)
  • Received : 2019.11.20
  • Accepted : 2020.03.22
  • Published : 2020.04.30
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Abstract

Leishmaniasis is a prevalent cause of death and animal morbidity in underdeveloped countries of endemic area. However, there is few vaccine and effective drugs. Antimicrobial peptides are involved in the innate immune response in many organisms and are being developed as novel drugs against parasitic infections. In the present study, we synthesized a 5-amino acid peptide REDLK, which mutated the C-terminus of Pseudomonas exotoxin, to identify its effect on the Leishmania tarentolae. Promastigotes were incubated with different concentration of REDLK peptide, and the viability of parasite was assessed using MTT and Trypan blue dye. Morphologic damage of Leishmania was analyzed by light and electron microscopy. Cellular apoptosis was observed using the annexin V-FITC/PI apoptosis detection kit, mitochondrial membrane potential assay kit and flow cytometry. Our results showed that Leishmania tarentolae was susceptible to REDLK in a dose-dependent manner, disrupt the surface membrane integrity and caused parasite apoptosis. In our study, we demonstrated the leishmanicidal activity of an antimicrobial peptide REDLK from Pseudomonas aeruginosa against Leishmania tarentolae in vitro and present a foundation for further research of anti-leishmanial drugs.

Keywords

References

  1. Desjeux P. Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 2014; 27: 305-318. https://doi.org/10.1016/j.cimid.2004.03.004
  2. Castillo-Ureta H, Zazueta-Moreno JM, Rendon-Maldonado JG, Torres-Avendano JI, Lopez-Moreno HS, Olimon-Andalon V, Salomon-Soto VM, Perez-Sanchez FP, Torres-Montoya EH. First report of autochthonous canine leishmaniasis caused by Leishmania (L.) mexicana in Sinaloa, Mexico. Acta Trop 2018; 190: 253-256. https://doi.org/10.1016/j.actatropica.2018.11.027
  3. Cizmeci Z, Karakusb M, Karabelac SN, Erdogand B, Guleca N. Leishmaniasis in Istanbul: A new epidemiological data about refugee leishmaniasis. Acta Trop 2019; 195: 23-27. https://doi.org/10.1016/j.actatropica.2019.04.008
  4. Aronson NE, Joya CA. Cutaneous leishmaniasis updates in diagnosis and management. Infect Dis Clin North Am 2019; 33: 101-117. https://doi.org/10.1016/j.idc.2018.10.004
  5. Griensven J, Diro E. Visceral leishmaniasis: recent advances in diagnostics and treatment regimens. Infect Dis Clin North Am 2019; 33: 79-99. https://doi.org/10.1016/j.idc.2018.10.005
  6. Kapil S, Singh PK, Silakari O. An update on small molecule strategies targeting leisghmaniasis. Eur J Med Chem 2018; 157: 339-367. https://doi.org/10.1016/j.ejmech.2018.08.012
  7. Marty P, Rosenthal, E. Treatment of visceral leishmaniasis: a review of current treatment practices. Expert Opin Pharmacother 2002; 3: 1101-1108. https://doi.org/10.1517/14656566.3.8.1101
  8. AlMatar M, Makky EA, Yakici G, Var I, Kayar B, Koksal F. Antimicrobial peptides as an alternative to anti-tuberculosis drugs. Pharmacol Res 2017; 128: 288-305. https://doi.org/10.1016/j.phrs.2017.10.011
  9. Ebenhan T, Gheysens O, Kruger HG, Zeevaart JR, Sathekge MM. Antimicrobial peptides: their role as infection-selective tracers for molecular imaging. Biomed Res Int 2014; 867381. https://doi.org/10.1155/2014/867381
  10. Lynn MA, Kindrachuk J, Marr AK, Jenssen H, Pante N, Elliott MR, Napper E, Hancock RR, Master WRM. Effect of BMAP-28 antimicrobial peptides on Leishmania major promastigote and amastigote growth: role of leishmanolysin in parasite survival. PLoS Negl Trop Dis 2011; 5: e1141. https://doi.org/10.1371/journal.pntd.0001141
  11. Aguilar-Diaz H, Canizalez-Roman A, Nepomuceno-Mejia T, Gallardo-Vera F, Hornelas-Orozco Y, Nazmi K, Bolscher JG, Carrero JC, Leon-Sicairos C, Leon-Sicairos N.Parasiticidal effect of synthetic bovine lactoferrin peptides on the enteric parasite Giardia intestinalis. Biochem Cell Biol 2017; 95: 82-90. https://doi.org/10.1139/bcb-2016-0079
  12. Cederlund A, Gudmundsson GH, Agerberth B. Antimicrobial peptides important in innate immunity. FEBS J 2011; 278: 3942-3951. https://doi.org/10.1111/j.1742-4658.2011.08302.x
  13. Muller F, Cunningham T, Beers R, Bera TK, Wayne AS, Pastan I. Domain II of Pseudomonas exotoxin is critical for efficacy of bolus doses in a xenograft model of acute lymphoblastic leukemia. Toxins 2018; 10: E210.
  14. Qaiser H, Aslam F, Iftikhar S, Farooq A. Construction and recombinant expression of Pseudomonas aeruginosa truncated exotoxin A in Escherichia coli. Cell Mol Biol (Noisy-le-grand) 2018; 64: 64-69.
  15. Chaudhary VK, Jinno Y, Gallo MG, FitzGerald D, Pastan I. Mutagenesis of Pseudomonas exotoxin in identification of sequences responsible for the animal toxicity. J Biol Chem 1990; 265: 16306-16310. https://doi.org/10.1016/S0021-9258(17)46223-5
  16. Bolhassani A, Taheri T, Taslimi Y, Zamanilui S, Zahedifard F, Seyed N, Torkashvand F, Vaziri B, Rafati S. Fluorescent Leishmania species: development of stable GFP expression and its application for in vitro and in vivo studies. Exp Parasitol 2011; 127: 637-645. https://doi.org/10.1016/j.exppara.2010.12.006
  17. Hansen PR, Oddo A. Fmoc solid-phase peptide synthesis. Methods Mol. Biol 2015; 1348: 33-50. https://doi.org/10.1007/978-1-4939-2999-3_5
  18. Kiderlen AF, Kaye PM. A modified colorimetric assay of macrophage activation for intracellular cytotoxicity against Leishmania parasites. J Immunol Methods 1990; 127: 11-18. https://doi.org/10.1016/0022-1759(90)90334-R
  19. McGwire BS, Olson CL, Tack BF, Engman DM. Killing of African trypanosomes by antimicrobial peptides. J Infect Dis 2003; 188: 146-152. https://doi.org/10.1086/375747
  20. Zhang X, Liu X, Shang HF, Xu Y, Qian MZ. Monocyte chemoattractant protein-1 induces endothelial cell apoptosis in vitro through a p53-dependent mitochondrial pathway. Acta Biochim Biophys Sin 2011; 43: 787-795. https://doi.org/10.1093/abbs/gmr072
  21. Tang CL, Liang J, Qian JF, Jin LP, Du MR, Li MQ, Li DJ. Opposing role of JNK-p38 kinase and ERK1/2 in hydrogen peroxide-induced oxidative damage of human trophoblast-like JEG-3 cells. Int J Clin Exp Pathol 2014; 7: 959-968.
  22. Sacks DL, Perkins PV. Identification of an infective stage of Leishmania promastigotes. Science 1984; 223: 1417-1419. https://doi.org/10.1126/science.6701528
  23. de Sousa APS, de Oliveira SSC, d'Avila-Levy CM, Dos Santos ALS, Branquinha MH. Susceptibility of promastigotes and intracellular amastigotes from distinct Leishmania species to the calpain inhibitor MDL28170. Parasitol Res 2018; 117: 2085-2094. https://doi.org/10.1007/s00436-018-5894-7
  24. Raymond F, Boisvert S, Roy G, Ritt JF, Legare D, Isnard A, Stanke M, Olivier M, Tremblay MJ, Papadopoulou B, Ouellette M, Corbeil J. Genome sequencing of the lizard parasite Leishmania tarentolae reveals loss of genes associated to the intracellular stage of human pathogenic species. Nucleic Acids Res 2012; 40: 1131-1147. https://doi.org/10.1093/nar/gkr834
  25. Taylor VM, Munoz DL, Cedeno DL, Velez ID, Jones MA, Robledo SM. Leishmania tarentolae: utility as an in vitro model for screening of antileishmanial agents. Exp Parasitol 2010; 126: 471-475. https://doi.org/10.1016/j.exppara.2010.05.016
  26. Lohner K, Blondelle SE. Molecular mechanisms of membrane perturbation by antimicrobial peptides and the use of biophysical studies in the design of novel peptide antibiotics. Comb Chem High Throughput Screen 2005; 8: 241-256. https://doi.org/10.2174/1386207053764576
  27. Eaton P, Bittencourt BSc CR, Silva VC, VerasMSc LMC, Costa CHN, Feio MJ, Leite JRSA. Anti-leishmanial activity of the antimicrobial peptide DRS 01 observed in Leishmania infantum (syn. Leishmania chagasi) cells. Nanomedicine 2014; 10: 483-490. https://doi.org/10.1016/j.nano.2013.09.003
  28. khalili S, Ebrahimzade E, Mohebali M, Shayan P, Mohammadi-Yeganeh S, Moghaddam MM, Elikaee S, Akhoundi B, Sharifi-Yazdi MK. Investigation of the antimicrobial activity of a short cationic peptide against promastigote and amastigote forms of Leishmania major (MHRO/IR/75/ER): an in vitro study. Exp Parasitol 2019; 196: 48-54. https://doi.org/10.1016/j.exppara.2018.11.006
  29. Torrent M, Pulido D, Rivas L, Andreu D. Antimicrobial peptide action on parasites. Curr Drug Targets 2012; 13: 1138-1147. https://doi.org/10.2174/138945012802002393
  30. Lili C, Weina J, Songgao C, Panpan Z, Juan L, Hang D, Yanbing G, Quan L, Pengtao G. In vitro leishmanicidal activity of antimicrobial peptide KDEL against Leishmania tarentolae. Acta Bioch Bioph Sin 2019; 51: 1286-1292. https://doi.org/10.1093/abbs/gmz128
  31. Hassani K, Shio MT, Martel C, Faubert D, Olivier M. Absence of metalloprotease GP63 alters the protein content of Leishmania exosomes. PLoS One 2014; 9: e95007. https://doi.org/10.1371/journal.pone.0095007
  32. Parashar S, Mukhopadhyay A. GTPase Sar1 regulates the trafficking and secretion of the virulence factor gp63 in Leishmania. J Biol Chem 2017; 292: 12111-12125. https://doi.org/10.1074/jbc.M117.784033
  33. Risso A, Braidot E, Sordano MC. BMAP-28, an antibiotic peptide of innate immunity, induces cell death through opening of the mitochondrial permeability transition pore. Mol Cell Biol 2002; 22: 1926-1935. https://doi.org/10.1128/MCB.22.6.1926-1935.2002
  34. Arayan LT, Kim HB, Reyes AWB, Huy NTX, Hong IH, Lee K, Yeom J, Park Y, Kim S. The immunomodulatory effect of antimicrobial peptide HPA3P restricts Brucella abortus 544 infection in BALB/c mice. Vet Microbiol 2018; 225: 17-24. https://doi.org/10.1016/j.vetmic.2018.09.005
  35. Bowdish DM, Davidson DJ, Scott MG, Hancock RE. Immunomodulatory activities of small host defense peptides. Antimicrob Agents Chemother 2005; 49: 1727-1732. https://doi.org/10.1128/AAC.49.5.1727-1732.2005