The kiss of (cell) death: can venom-induced immune response contribute to dermal necrosis following arthropod envenomations?

Introduction: Snakes, insects, arachnids and myriapods have been linked to necrosis following envenomation. However, the pathways involved in arthropod venom-induced necrosis remain a highly controversial topic among toxinologists, clinicians and the public. On the one hand, clinicians report on alleged envenomations based on symptoms and the victims' information. On the other hand, toxinologists and zoologists argue that symptoms are incompatible with the known venom activity of target species. This review draws from the literature on arthropod envenomations, snakebite, and inflammatory processes to suggest that envenomation by a range of organisms might trigger an intense inflammatory cascade that ultimately lead to necrosis. If confirmed, these processes would have important implications for the treatment of venom-induced necrosis. Objectives: To describe two inflammatory pathways of regulated necrosis, tumour necrosis factor (necroptosis) and Neutrophil Extracellular Traps (NETosis); to discuss existing knowledge about snake venom and arachnid-induced necrosis demonstrating the involvement of tumour necrosis factor and neutrophils in the development of tissue necrosis following envenomation and to contribute to the understanding of venom-induced necrosis by arthropods and provide clinicians with an insight into little known inflammatory processes which may occur post envenomation. Methods: ISI Web of Science databases were searched using the terms "spider bite necrosis", "arthropod envenomation necrosis", "venom necrosis", "venom immune response", "loxoscelism", "arachnidism", "necroptosis venom", "necroptosis dermatitis", "tumour necrosis factor TNF venom", "scorpionism", "scolopendrism", "centipede necrosis", "NETosis venom", "NETosis necrosis". Searches produced 1737 non-duplicate citations of which 74 were considered relevant to this manuscript. Non-peer-reviewed sources or absence of voucher material identifying the organism were excluded. What is necrosis? Necrosis is the breakdown of cell membrane integrity followed by inflowing extracellular fluid, organelle swelling and the release of proteolytic enzymes into the cytosol. Necrosis was historically considered an unregulated process; however, recent studies demonstrate that necrosis can also be a programmed event resulting from a controlled immune response (necroptosis). Tumour necrosis factor and the necroptosis pathway: Tumour necrosis factor is a pro-inflammatory cytokine involved in regulating immune response, inflammation and cell death/survival. The pro-inflammatory cytokine TNF-α participates in the development of necrosis after envenomation by vipers. Treatment with TNF-α-antibodies may significantly reduce the manifestation of necrosis. Neutrophil Extracellular Traps and the NETosis pathway: The process by which neutrophils discharge a mesh of DNA strands in the extracellular matrix to entangle ("trap") pathogens, preventing them from disseminating. Neutrophil Extracellular Traps have been recently described as important in venom-induced necrosis. Trapped venom accumulates at the bite site, resulting in significant localized necrosis. Arthropod venom driving necrosis: Insects, myriapods and arachnids can induce necrosis following envenomation. So far, the processes involved have only been investigated in two arachnids: Loxosceles spp. (recluse spiders) and Hemiscorpius lepturus (scorpion). Loxosceles venom contains phospholipases D which hydrolyse sphingomyelin, resulting in lysis of muscle fibers. Subsequently liberated ceramides act as intermediaries that regulate TNF-α and recruit neutrophils. Experiments show that immune-deficient mice injected with Loxosceles venom experience less venom-induced inflammatory response and survive longer than control mice. Necrosis following Hemiscorpius lepturus stings correlates with elevated concentrations of TNF-α. These observations suggest that necrosis may be indirectly triggered or worsened by pathways of regulated necrosis in addition to necrotic venom compounds. Conclusions: Envenomation often induce an intense inflammatory cascade, which under certain circumstances may produce necrotic lesions independently from direct venom activity. This could explain the inconsistent and circumstantial occurrence of necrosis following envenomation by a range of organisms. Future research should focus on identifying pathways to regulated necrosis following envenomation and determining more efficient ways to manage inflammation. We suggest that clinicians should consider the victim's immune response as an integral part of the envenomation syndrome.

[Biological properties of heat-labile lethal toxin of Yersinia pseudotuberculosis].

The aim of the study was to assess the biological properties of heat-labile lethal protein toxin of Y. pseudotuberculosis. Toxin was extracted from Y. pseudotuberculosis strain 2517 type III serovar pYV-. The toxin killed mice 1-3 days after intraperitoneal administration (LD50=0.3 mcg of the protein). Heating at 56 degrees C during 30 min inactivated lethal activity of the toxin. It had a dose-dependent dermonecrotic effect during intracutaneous administration in rabbits. Hyperimmune rabbit serum to the toxin was obtained. Incubation of the toxin (LD100=1.2 mcg of the protein) with the serum at 37 degrees C during 30 min resulted in neutralization of lethal and dermonecrotic effects. The toxin did not have the cytotoxic effect on HeLa, Hep-2, and SPEV cells, but showed hemolytic activity to human and animal erythrocytes, and weak mitogenic activity to splenic cells of CBA line mice compared with control mitogen (concanavalin A).

[The properties of the Vibrio cholerae strains isolated in large districts in western Dagestan in 1994]. / Svoistva shtammov Vibrio cholerae, vydelennykh v krupnykh raionakh na zapade Dagestana v 1994 g.

190 V. cholerae cultures isolated by the specialized antiepidemic brigade of the Rostov-on-Don Research Institute for Plague Control in the Khasavyurt, Babayurt and Novolaksk regions of Daghestan in August-October 1994. All isolated strains were typical with respect to their morphological and cultural properties and could be agglutinated (with the exception of one strain) to the titer or half-titer with diagnostic cholera serum and Ogawa serum. 4 strains had signs of RO-dissociation, 4 strains were agglutinated with Inaba serum in a low titer. All strains were resistant to diagnostic bacteriophages. Cyproxin and doxicycline proved to be the most active agents for the treatment of patients. Agglutinins, vibriocidins and antidermonectrotic antibodies in diagnostic titers were detected in the sera of all patients and Vibrio carriers.

Dermonecrotic and lethal components of Loxosceles gaucho spider venom.

Loxosceles gaucho spider venom causes a typical dermonecrotic lesion in bitten patients and rarely causes lethal systemic effects. Gel filtration on Sephadex G 100 of L. gaucho spider venom resulted in three fractions: fraction A, containing the higher mol. wt components (approximately 35,000); fraction B, containing lower mol. wt components (approximately 15,000); and fraction C, containing very low mol. wt components (probably small peptides). The dermonecrotic and lethal activities were detected exclusively in fraction A. The venom and fraction A produced large dermonecrotic lesions in rabbits with necrosis spreading by gravity to the skin of the lateral body wall. Analysis by SDS-PAGE showed that the proteins contained in fraction A are approximately 35,000 and 33,000 mol. wt. Immunoblotting analysis showed that the proteins responsible for the dermonecrotic and lethal activity are very immunogenic and the first to be detected by antibodies during the course of immunization.

[Isolation and certain properties of the dermonecrotic toxin from Pasteurella multocida]. / Vydelenie i nekotorye svoistva dermonekroticheskogo toksina Pasteurella multocida.

The procedure for isolation and purification of Pasteurella multocida serovariant D toxin has been described. It includes the three steps of protein precipitation from cultural filtrates by 70% ammonium sulfate, chromatography of the concentrated material on Ultragel AcA44 gel-filtration on Sephracryl S-200. The proposed technique permits one the 155-fold purification of the preparation with 32.6% yield estimated by biological activity. The obtained purified preparation is homogeneous in polyacrylamide gel electrophoresis. The immunological methods also confirm the homogeneity of the preparation. The minimal dermonecrotic dose for guinea pigs of the purified 120 kDa toxin is 78 ng and LD50 for mice is 280 ng. Pasteurella multocida toxin is found to be a thermolabile protein sensitive to trypsin, glutaraldehyde and formaldehyde treatments.

Antigenic relationship between the dermonecrotic toxins produced by Pasteurella multocida type D and type A.

Crude dermonecrotic toxins (DNT) were prepared from Pasteurella multocida (P.m.) type D and type A strains isolated from pigs with atrophic rhinitis. Rabbits were immunized with the DNT of P.m. type D. This serum neutralized the DNT of P.m. type A to the same degree as the homologous one both in vitro (cytopathogenicity for tissue culture cells) and in vivo (mouse lethality and dermonecrotic activity in guinea pig).

Effects of Bordetella pertussis components on IgE and IgG1 responses.

The effect of dermonecrotic toxin (DNT), fimbrial hemagglutinin (FHA), K-agglutinogen, lipopolysaccharide (LPS), and pertussigen from Bordetella pertussis on the production of IgE and IgG1 antibodies to hen egg albumin (Ea) was investigated in C57BL/6 mice. The IgE antibody contents were determined by passive cutaneous anaphylaxis (PCA) in the skin of Lewis rats, while the IgG1 antibody contents were determined by PCA reactions on the skin of mice using sera that had been heated for 3 hr at 56 C to destroy the IgE antibodies. Among the B. pertussis components tested, pertussigen was the most effective adjuvant for increasing the IgE and IgG1 antibodies to Ea. LPS also moderately increased both types of antibodies, and FHA slightly increased the IgG1 titers. When LPS was given 5 days before Ea, it suppressed both IgE and IgG1 titers while FHA had only slight adjuvant action on both type of antibodies. When each of the components was tested for its ability to modify the adjuvant action of pertussigen, it was found that only DNT interfered significantly with the adjuvanticity of pertussigen when given on the day of immunization with Ea. When the components were given 5 days before Ea, DNT produced significant suppression of only the IgG1 response. LPS, FHA, and K-agglutinogen did not significantly affect the adjuvant action of pertussigen.