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Identification, Characterization, Immunolocalization, and Biological Activity of Lucilin Peptide

2018; Elsevier BV; Volume: 185; Linguagem: Inglês

10.1016/j.actatropica.2018.06.003

1873-6254

Germán Alberto Téllez Ramírez, Jesica Alejandra Zapata, Lily Johanna Toro, Diana Carolina Toro Henao, Juan Pablo Bedoya Agudelo, Juan David Rivera, Juan Valentin Trujillo, Bruno Rivas‐Santiago, Richard Hoyos–López, Jhon Carlos Castaño Osório,

Insect and Pesticide Research

Maggots from the Lucilia sp. genus are used for debridement of infected and necrotic wounds. Broad-spectrum antimicrobial activity has been described in the excretion/secretions (ES1) of these larvae. This study identifies the genetic sequence of a cecropin-like antimicrobial peptide from Lucilia eximia. Total RNA was extracted and used for PCR-RACE amplification of a cecropin, the native peptide was immunolocalized in the tissues and secretions of the larvae, and a synthetic analog was used to explore its antimicrobial, cytotoxic, LPS neutralizing and wound-healing activities in vitro. The genetic cDNA sequence of a cecropin-like antimicrobial peptide in L. eximia called "Lucilin" was amplified, corresponding to 63 aa completed protein and 40 aa mature peptide; the structure of the mature peptide was predicted as an α-helix. The peptide was immunolocalized in the salivary glands, fat body, the ES, and hemolymph of the maggots. Lucilin synthetic peptide analog was active against E. coli DH10B with a MIC2 of 7.8 μg/mL, E. coli extended spectrum b-lactamase (ESBL) (MIC: 15.6 μg/mL), and Enterobacter cloacae (MIC: 125 μg/mL), but it was not active against Pseudomonas aeruginosa and Staphylococcus epidermidis; and had no cytotoxic or hemolytic activity. It showed immunomodulatory activity against human peripheral blood mononuclear cells (PBMCs) stimulated with LPS, reducing the TNF-α production when treated at 17 μg/mL and induces cell migration of Hacat at 5 and 50 μg/mL. Lucilin is a cecropin-like peptide from L. eximia with antimicrobial activity against Gram negative bacteria and immunomodulatory activities, decreasing the TNF-α production in PBMCs and inducing cellular migration in human keratinocytes.