title: Phospholipase D from Loxosceles laeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration.
abstract: Cutaneous loxoscelism envenomation byLoxoscelesspiders is characterized by the development of a dermonecrotic lesion, strong inflammatory response, the production of pro-inflammatory mediators, and leukocyte migration to the bite site. The role of phospholipase D (PLD) fromLoxoscelesin the recruitment and migration of monocytes to the envenomation site has not yet been described. This study reports on the expression and production profiles of cytokines and chemokines in human skin fibroblasts treated with catalytically active and inactive recombinant PLDs fromLoxosceles laeta(rLlPLD) and lipid inflammatory mediators ceramide 1-phosphate (C1P) and lysophosphatidic acid (LPA), and the evaluation of their roles in monocyte migration. Recombinant rLlPLD1 (active) and rLlPLD2 (inactive) isoforms induce interleukin (IL)-6, IL-8, CXCL1/GRO-α, and CCL2/monocyte chemoattractant protein-1 (MCP-1) expression and secretion in fibroblasts. Meanwhile, C1P and LPA only exhibited a minor effect on the expression and secretion of these cytokines and chemokines. Moreover, neutralization of both enzymes with anti-rLlPLD1 antibodies completely inhibited the secretion of these cytokines and chemokines. Importantly, conditioned media from fibroblasts, treated with rLlPLDs, stimulated the transmigration of THP-1 monocytes. Our data demonstrate the direct role of PLDs in chemotactic mediator synthesis for monocytes in human skin fibroblasts and indicate that inflammatory processes play an important role during loxoscelism.
authors.affiliation: Laboratorio de Parasitología Molecular, Departamento de Tecnología Médica, Facultad de Ciencias de la Salud, Universidad de Antofagasta, Antofagasta, CP 1270300, Chile. jose.rojas@uantof.cl.
authors.forename: José M
authors.initials: JM
authors.lastname: Rojas
authors.affiliation: Laboratorio de Parasitología Molecular, Departamento de Tecnología Médica, Facultad de Ciencias de la Salud, Universidad de Antofagasta, Antofagasta, CP 1270300, Chile. tomas.aran@uantof.cl.
authors.forename: Tomás
authors.initials: T
authors.lastname: Arán-Sekul
authors.affiliation: Laboratorio de Parasitología Molecular, Departamento de Tecnología Médica, Facultad de Ciencias de la Salud, Universidad de Antofagasta, Antofagasta, CP 1270300, Chile. emmanuel_cr@hotmail.cl.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Cortés
authors.affiliation: Laboratorio de Parasitología Molecular, Departamento de Tecnología Médica, Facultad de Ciencias de la Salud, Universidad de Antofagasta, Antofagasta, CP 1270300, Chile. romyjaldin@gmail.com.
authors.forename: Romina
authors.initials: R
authors.lastname: Jaldín
authors.affiliation: Laboratorio de Parasitología Molecular, Departamento de Tecnología Médica, Facultad de Ciencias de la Salud, Universidad de Antofagasta, Antofagasta, CP 1270300, Chile. kely.ordenes@uantof.cl.
authors.forename: Kely
authors.initials: K
authors.lastname: Ordenes
authors.affiliation: Departamento Biomédico, Facultad de Ciencias de la Salud, Universidad de Antofagasta, Antofagasta, CP 1270300, Chile. patricio.orrego@uantof.cl.
authors.forename: Patricio R
authors.initials: PR
authors.lastname: Orrego
authors.affiliation: Laboratorio de Parasitología Molecular, Departamento de Tecnología Médica, Facultad de Ciencias de la Salud, Universidad de Antofagasta, Antofagasta, CP 1270300, Chile. jorge.gonzalez@uantof.cl.
authors.forename: Jorge
authors.initials: J
authors.lastname: González
authors.affiliation: Laboratorio de Parasitología Molecular, Departamento de Tecnología Médica, Facultad de Ciencias de la Salud, Universidad de Antofagasta, Antofagasta, CP 1270300, Chile. jearoj@yahoo.com.
authors.forename: Jorge E
authors.initials: JE
authors.lastname: Araya
authors.affiliation: Laboratorio de Parasitología Molecular, Departamento de Tecnología Médica, Facultad de Ciencias de la Salud, Universidad de Antofagasta, Antofagasta, CP 1270300, Chile. alejandro.catalan@uantof.cl.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Catalán
mesh_terms: D000818:Animals; D060829:Arthropod Proteins / Q000494:pharmacology*; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D002518:Ceramides / Q000494:pharmacology; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008246:Lysophospholipids / Q000494:pharmacology; D009000:Monocytes / Q000187:drug effects* / Q000502:physiology; D010739:Phospholipase D / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D012867:Skin / Q000166:cytology; D013111:Spider Venoms / Q000201:enzymology*; D013112:Spiders
chemical_list: D060829:Arthropod Proteins; D002518:Ceramides; D016207:Cytokines; D008246:Lysophospholipids; D012333:RNA, Messenger; D011994:Recombinant Proteins; D013111:Spider Venoms; C065576:ceramide 1-phosphate; D010739:Phospholipase D; C032881:lysophosphatidic acid
keywords: Loxosceles laeta; chemokines; monocytes migration; phospholipase D