title: Temporizin and Temporizin-1 Peptides as Novel Candidates for Eliminating Trypanosoma cruzi.
abstract: Tropical diseases caused by parasitic infections continue to cause socioeconomic distress worldwide. Among these, Chagas disease has become a great concern because of globalization. Caused by Trypanosoma cruzi, there is an increasing need to discover new, more effective methods to manage infections that minimize disease onset. Antimicrobial peptides represent a possible solution to this challenge. As effector molecules of the innate immune response against pathogens, they are the first line of defense found in all multi-cellular organisms. In amphibians, temporins are a large family of antimicrobial peptides found in skin secretions. Their functional roles and modes of action present unique properties that indicate possible candidates for therapeutic applications. Here, we investigated the trypanocide activity of temporizin and temporizin-1. Temporizin is an artificial, hybrid peptide containing the N-terminal region of temporin A, the pore-forming region of gramicidin and a C-terminus consisting of alternating leucine and lysine. Temporizin-1 is a modification of temporizin with a reduction in the region responsible for insertion into membranes. Their activities were evaluated in a cell permeabilization assay by flow cytometry, an LDH release assay, electron microscopy, an MTT assay and patch clamp experiments. Both temporizin and temporizin-1 demonstrated toxicity against T. cruzi with temporizin displaying slightly more potency. At concentrations up to 100 μg/ ml, both peptides exhibited low toxicity in J774 cells, a macrophage lineage cell line, and no toxicity was observed in mouse primary peritoneal macrophages. In contrast, the peptides showed some toxicity in rat adenoma GH3 cells and Jurkat human lymphoma cells with temporizin-1 displaying lower toxicity. In summary, a shortened form of the hybrid temporizin peptide, temporizin-1, was efficient at killing T. cruzi and it has low toxicity in wild-type mammalian cells. These data suggest that temporizin-1 might be a candidate for Chagas disease therapy.
authors.affiliation: FIOCRUZ, Center for Technological Development in Health (CDTS)/National Institute of Science and Technology for Innovation on Neglected Diseases (INCT-IDN), Rio de Janeiro, RJ, Brazil.|FIOCRUZ, Oswaldo Cruz Institute, Laboratory of Experimental and Computational Biochemistry of Pharmaceuticals, Rio de Janeiro, RJ, Brazil.
authors.forename: André L A
authors.initials: AL
authors.lastname: Souza
authors.affiliation: FIOCRUZ, Oswaldo Cruz Institute, Laboratory of Toxoplasmosis and other Protozoosis, Rio de Janeiro, RJ, Brazil.
authors.forename: Robson X
authors.initials: RX
authors.lastname: Faria
authors.affiliation: FIOCRUZ, Oswaldo Cruz Institute, Laboratory of Imunomodulation and Protozoology, Rio de Janeiro, RJ, Brazil.
authors.forename: Kátia S
authors.initials: KS
authors.lastname: Calabrese
authors.affiliation: FIOCRUZ, Oswaldo Cruz Institute, Laboratory of Imunomodulation and Protozoology, Rio de Janeiro, RJ, Brazil.
authors.forename: Daiane J
authors.initials: DJ
authors.lastname: Hardoim
authors.affiliation: FIOCRUZ, Adolfo Lutz Institute, Electronic Microscopy section, Araçatuba, São Paulo, SP, Brazil.
authors.forename: Noemi
authors.initials: N
authors.lastname: Taniwaki
authors.affiliation: FIOCRUZ, Oswaldo Cruz Institute, Laboratory of Cellular Communication, Rio de Janeiro, RJ, Brazil.
authors.forename: Luiz A
authors.initials: LA
authors.lastname: Alves
authors.affiliation: FIOCRUZ, Center for Technological Development in Health (CDTS)/National Institute of Science and Technology for Innovation on Neglected Diseases (INCT-IDN), Rio de Janeiro, RJ, Brazil.|Federal Fluminense University, Biology Institute, Department of Cellular and Molecular Biology, Niterói, RJ, Brazil.
authors.forename: Salvatore G
authors.initials: SG
authors.lastname: De Simone
mesh_terms: D000818:Animals; D023181:Antimicrobial Cationic Peptides; D049109:Cell Proliferation; D014355:Chagas Disease / Q000188:drug therapy* / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D019169:Jurkat Cells; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects; D008297:Male; D051379:Mice; D008854:Microscopy, Electron; D010455:Peptides / Q000494:pharmacology*; D011506:Proteins / Q000494:pharmacology*; D014349:Trypanosoma cruzi / Q000276:immunology*
chemical_list: D023181:Antimicrobial Cationic Peptides; D010455:Peptides; D011506:Proteins; C104501:temporin; C000621582:temporizin-1 peptide; D007770:L-Lactate Dehydrogenase