Imunogenicidade do veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae)

Abstract

CHAPTER II: The Dinoponera presents six species, all with distribution by South America. These ants are characterized by the presence of a venom gland, which is responsible for the production of a toxin, consists for the most part by proteins. This work was aimed at the identification of specific peptides of the venom of D. australis by Phage Display. It was used commercial library of random peptides Ph.D.-12 Peptide 12-mer Phage Display Library. After four stages of selection, 48 clones had their DNA sequence determined and have been translated, generating 26 valid sequences and 16 different peptides. Analyses of bioinformatics identified similarity with ADAM metaloprotease of Culex quinquefasciatus and with a protein similar to acid phosphatase of venom found in Nasonia vitripennis. ADAM metalloproteases are proteins that act in control of accession and of membrane fusion, cytokine and growth factor. Acid Phosphatase is one of the largest of invertebrates Lysosomal enzymes associated with degradation and cell death. The activity of Phosphatase acid in this ant venom was made using the kit Api Zym. The mapping of the components of the venom of D. australis, showed that the peptides tested not have similarity to: mellitin of Apis mellifera and Vespa magnifica; largest allergen of Myrmecia pilosula; Apamin of A. mellifera; phospholipase A2 from A. melífera, but showed similarity with phospholipase A2 from Nasonia vitripennis. He also similarity with different allergens, neurotoxinas, metalloproteases, fenoloxidase and hialuronidase of different hymenopters. To examine the effect of peptides used if the test edematogênico noting that the immunized mice showed higher edematogênico level with 30 minutes and therefore faster than mice not immunized. CHAPTER III: The analysis of the components of the venom of himenopteras is a rich source of information on the functions and mechanisms of biological poison. It is important, also, for trigger allergic reactions in humans and to present therapeutic applications. The cytokines are a heterogeneous group of mediators cellular peptídicos associated with activation of the immune system and inflammatory responses. The Toll-like receptors (TLRs) are proteins transmembranas characterized by repeated sequences extracellular leucine-rich, capable of recognizing a broad group of pathogens associated with the molecular patterns (PAMPs) of different species of microorganisms and essential for the activation of the innate immune response. This study examined the profile of immune response in blood cells in vitro, stimulated by venom of Dinoponera australis in different concentrations (0.5, 1 and 3 mg) and examined, also, the effect of the same concentrations of poisonous raw in the expression of mRNA the gene TLR4 in peripheral blood mononuclear cells (PBMC) by Real Time PCR. Human blood was obtained from volunteer donors, collected in tubes containing heparin. The cytokines in culture supernatants of peripheral blood were measured by ELISA using pairs of monoclonal antibodies commercially available. The venom at all concentrations tested significantly stimulated the production of IL-10 in culture of whole blood and reduction of TNF-α. The IL-10 has a high anti-inflammatory activity both in the function of lymphocytes as monocytes. This decreases the cytokine production of a series of pro-inflammatory cytokines as IL1, TNF-α and IL6. The levels of gene expression of TRL4 were determined by real-time PCR and normalized to the levels of expression of the gene constituent HPRT. There was a significant decrease in the expression of mRNA of the gene TLR4 in culture of human PBMC only to the concentration of 3 g μ These data suggest that some component of crude that ant poison takes effect in modulating the expression of TLR4 and potent antiinflammatory effect.