Identification and evaluation of Galleria mellonella peptides with antileishmanial activity

• Published in: Analytical biochemistry Vol. 546; pp. 35 - 42
• Main Authors: Patiño-Márquez, Isabel A., Patiño-González, Edwin, Hernández-Villa, Laura, Ortíz-Reyes, Blanca, Manrique-Moreno, Marcela
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.04.2018
• Subjects: Animals; Antimicrobial Cationic Peptides - chemistry; Antimicrobial Cationic Peptides - isolation & purification; Antimicrobial Cationic Peptides - pharmacology; Antimicrobial peptides; Antiprotozoal Agents - chemistry; Antiprotozoal Agents - isolation & purification; Antiprotozoal Agents - pharmacology; Electrophoresis; Galleria mellonella; Hemolymph; Leishmania; Leishmania - drug effects; Lepidoptera - chemistry; Parasitic Sensitivity Tests; Reverse phase liquid chromatography; FDA; ABC; LPG; A. niger; DTT; SbV; MTT; TFA; AMPs; M. luteus; Antimicrobial peptides; Electrophoresis; D. melanogaster; E. coli; WHO; PBS; BCA; H. cecropia; S. lutea; Hemolymph; IAA; CL; G. mellonella; Galleria mellonella; IEF; ACN; RP-HPLC; 2D gel electrophoresis; SDS-PAGE; L. monocytogenes; LC-MS; Leishmania; VL; MCL; Reverse phase liquid chromatography
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2018.01.029

Leishmaniasis is a neglected disease, World Health Organization (WHO) declared it as high priority worldwide. Colombia is one of the 98 countries in which the disease caused more than 17.000 cases per year. There is a need to explore novel therapies to reduce the side effects of the current treatments. For this reason, this study was aimed to evaluate Galleria mellonella hemolymph for potential peptides with anti-parasitic activity. Larvae were challenged with Leishmania (V) panamensis promastigotes and hemolymph was analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), reversed-phase chromatography (RP-HPLC), two-dimensional gel electrophoresis and liquid chromatography-mass spectroscopy (LC/MS). The immunological response of Galleria mellonella was followed by SDS-PAGE, immunized hemolymph was fractionated by RP-HPLC where fractions 5 and 11 showed the highest antileishmanial activity. From these fractions 15 spots were isolated by 2D gel electrophoresis and evaluated by LC/MS to identify the peptides present in the spots. After the analysis Moricin-B, Moricin-C4, Cecropin-D and Anionic Peptide 2 were identified due to the immune challenge with Leishmania promastigotes. Anionic peptide 2 and Cecropin-D were synthesized and evaluated for antileishmanial activity. The results showed that Anionic peptide 2 presented more anti-parasitic activity. This study showed for the first time the anti-parasitic potential of peptides derived from hemolymph of Galleria mellonella. [Display omitted]