Snake venom detoxification methods -
Accordingly, it might seem plausible that targeting proinflammatory cytokines, chemokines, and small molecules or their receptors similarly represents novel therapeutic avenues for certain envenomations. However, research in this area is still in its infancy, and to date, studies have described both beneficial and detrimental outcomes of immunosuppression during experimental envenomation.
For example, the detection of snake and bee venom toxins by NOD-like receptor family, pyrin domain-containing 3 NLRP3 inflammasome, triggers immune cell activation, potent IL-1β secretion, and neutrophil influx Interestingly, Palm and Medzhitov showed that although inflammasome inhibition, such as seen in caspasedeficient mice, successfully inhibited cytokine release and leukocyte influx, it unexpectedly resulted in a higher susceptibility of the mice to the noxious effects of venoms, including mortality Conversely, Zoccal et al.
determined that using a hexapeptide ligand for the MΦ scavenger receptor CD36 protected mice against a lethal dose of T.
serrulatus scorpion venom through decreased production of IL-1β, IL-6, TNF, CCL3, and PGE2, and restrained lung inflammation While reduced IL-1β secretion and neutrophil influx was observed in both models, together, these data demonstrate the importance of innate immunodetection in the defence against bites and stings.
The primary effector function of B cells is the generation of antibodies immunoglobulins; Ig for humoral defence Bites, stings, and intentional venom inoculation stimulate the generation of venom protein-specific antibodies , Antibody-mediated neutralisation effectively counteracts venom activity However, a primary B cell response is slow requiring days to weeks to become fully active , while defence against rapid venom action requires an immediate response As an alternative to host antibodies, antivenom, produced in large mammals and purified for medical purposes, can provide passive immunity to victims of life-threatening envenomation — The following provides a simplified overview of a primary thymus-dependent humoral response towards envenomation.
Following bite or sting, APCs, such as DCs, MNCs and MΦs, capture and process venom proteins at the site of injury, promoting maturation , Matured APCs migrate to secondary lymphoid tissue to present venom antigen to naïve T H cells via membrane-bound peptide-MHC II protein complexes , In lymph nodes, engagement of a T cell receptor TCR with cognate peptide-MHC molecule initiates T H activation signal 1 Critical secondary signals, required for complete T cell activation, are provided by APCs.
APCs, especially DCs, highly express ligands including CD80 and CD86 for T cell co-stimulatory molecules, such as CD28 signal 2 Next, APC-derived and circulating cytokines as well as autocrine IL-2 induce T cell proliferation and differentiation signal 3.
During a primary antibody response, B cells require multiple stimulatory signals. The first occurs when a B cell receptor BCR encounters its specific soluble or membrane-bound epitope , The internalised antigen is processed and displayed on the B cell surface as a peptide-MHC complex for T H presentation , TCR binding triggers upregulation of co-stimulatory ligands, such as CD40L, and the production of proinflammatory cytokines, including IL-4 CD40L engagement with B cell CD40 mediates the recruitment of intracellular adaptor proteins essential for propagating downstream signalling Additionally, cytokines secreted by primed T H 2 cells provide B cells with accessory stimulation for the early proliferation and clonal expansion and later differentiation, antibody production, and isotype switching stages of B cell activation Proliferating B cells form germinal centres GCs where memory B and antibody-secreting plasma cells develop.
Here, B cells also undergo somatic hypermutation and isotype switching IgM and IgD to IgG, IgE, or IgA to generate high-affinity antibodies for robust immune responses Yet, critical though they may be, adaptive responses can also produce severe pathology For example, IgE isotype switching following venom challenge can, in a percentage of hypersensitive individuals, lead to fatal allergic reactions discussed below In addition to allergy, dysregulation of adaptive responses and loss of self-tolerance stimulate destructive auto-reactivity As such, lymphocytes T cells in particular are a target for therapeutic modulation Serendipitously, venoms can contain ligands for T cell ion channels and receptors, able to modulate immune function with high specificity described below — Cell-specific venom-mediated immune dysregulation is described above.
The following sections discuss modes of immunopathology, including venom-induced allergic reaction and systemic inflammation. Reflecting a global trend, honey bee A. mellifera stings are a significant contributor to venom injury in Australia, representing Venom from stinging Hymenopterans is commonly associated with allergic reactions worldwide In these incidences, systemic reactions SR are predominantly mediated by IgE-mechanisms; however, dose-dependent IgE-independent responses are also possible 7.
For honey bee venom, these hypersensitivity-inducing proteins include phospholipase A2, hyaluronidase, acid phosphatase, and dipeptidylpeptidase In vespid venom wasp and yellow jacket , Antigen 5 and phospholipase A1 are the recognised major allergens Classic IgE-mediated allergic disease begins with a sensitisation process.
Keratinocytes and resident immune cells detect damage induced by noxious substances, such as venom-derived compounds, stimulating the release of alarmins, cytokines IL-4, IL-5, and IL , and other proinflammatory mediators required for antibody production , DCs capture and process antigen for presentation to naïve T cells in draining lymph nodes, triggering events eventuating in plasma cell IgE antibody production , Elevated IgE is a normal physiological response following a bite or sting and is not necessarily predictive of disease 69 , 75 , Nevertheless, in some individuals, systemic IgE levels remain elevated longer term and can trigger SR, including anaphylactic shock, after multiple stings 7.
The symptoms of immediate Type-1 allergic reactions occur during secondary antigen challenges. When IgE encounters its cognate antigen, crosslinking of FcϵR1 on MCs and basophils in mucosal and epithelial tissues provoke activation and degranulation — Preformed inflammatory mediators, including histamine and proteases, are rapidly released from granules into the extracellular environment — Histamine is chiefly responsible for the clinical consequences of Type-1 allergic reactions , In allergic disease, histamine acting upon H 1 and H 2 receptors causes smooth muscle contraction, constriction of airways, swelling of the epiglottis, and increased vascular permeability.
These events lead to dangerously low blood pressure, oedema and potentially death , Further to this, proinflammatory genes, stimulated during the initial phase, induces de novo synthesis of the leukotrienes particularly LTB 4 , cytokines, and chemokines responsible for the late phase or delayed-type symptoms , These mediators are potent inducers of cell activation, migration, and the influx of lymphocytes and neutrophils , The incidence and severity of biphasic anaphylaxis are highly variable, and fatalities can occur, necessitating continued patient observation following the resolution of initial symptoms , , Adrenaline autoinjectors AAI are an essential first-line treatment; however, death may still occur despite prompt administration , Additional therapies include H 1 and H 2 antihistamines to counter the pathophysiological effects mediated through these receptors and critical supportive care , For individuals with verified IgE-mediated allergy, venom immunotherapy VIT may generate a lifesaving tolerance to known allergens The pathogenic role of MCs and IgE-mediated granule release is well established However, it has been postulated that allergy may be a barrier function disease in which cellular damage and perturbations of the epithelium and endothelium induce excessive proinflammatory responses from the resident immune cells If this hypothesis is correct, the therapeutic modulation of these cells may correct the imbalanced proinflammatory response but this hypothesis has not been investigated for venom-associated allergy.
The phylum Cnidaria classes Hydrozoa, Scyphozoa, and Cubozoa comprises approximately 10, jellyfish species distributed throughout the world , Jellyfish stings typically trigger local or large local responses, manifesting as pain, swelling, and erythema, but are usually not life-threatening , However, severe delayed cutaneous reaction, allergy, and anaphylactic shock can occur — It may not be surprising then that in the unexpected discovery of anaphylaxis by physiologists Charles Richet and Paul Portier involved marine venom from the Portuguese man-of-war Physalia physalis and sea anemone , When attempting to immunize dogs against harmful venom effects, Richet and Portier found that rather than confer protection phylaxis , a subsequent venom challenge resulted in death.
This discovery later won Richet the Nobel Prize in Physiology or Medicine , Jellyfish envenomation is the most common marine sting type, impacting fishers, surfers, and sea bathers globally.
An estimated million stings occur annually, with peak incidence coinciding with blooming or swarming seasons during warmer months A characteristic feature of the phylum Cnidaria is specialised stinging organelles, known as nematocysts. Located within cnidocytes, nematocysts are explosive capsule organelles containing coiled, barbed and threadlike tubules coated in venom High-velocity capsule release, triggered by physicochemical stimuli, causes inversion of tubules into harpoon-like threads able to puncture and penetrate prey and predators , Through nematocysts, jellyfish venom, containing pore-forming compounds, metalloproteases, serine proteases, and phospholipases, is injected into the victim, causing paralysis of prey and in humans dysregulation of immune function, cardiac function, respiratory function, and potentially fatal outcomes Although jellyfish venom can cause severe immediate-phase and delayed-type allergic reactions, the causative allergens are mostly unknown.
Recently, the N-linked glycoprotein, CqTX-A a hemolytic toxin , was identified as a major allergen from this venom While the underlying mechanism has yet to be elucidated, this finding has important implications as CqTX-A shares significant sequence homology with other lethal pore-forming jellyfish proteins, specifically, CfTX-1 and CfTX-2 Chironex fleckeri , CrTX-A Carybdea rastoni , CaTX-A Carybdea alata and CqTX-A Chiropsalmus quadrigatus Haeckel — For Hydrozoa which includes Portuguese man-of-war , Scyphozoa true jellyfish , and Cubozoa box jellyfish , nematocysts are located on the tentacles, oral arms, and in some instances, the bell of the jellyfish , Contact with tentacles can result in inoculation from potentially millions of nematocysts Once stung, venom distribution occurs via capillaries and the lymphatic system to target organs, while the barbed tubules remain embedded in the skin until clearance , Tubules are allergenic scaffolds comprising carbohydrates, proteins, chitin, and mini-collagen , , — As such, it has been suggested that impaired clearance, especially of chitin, may contribute to more severe outcomes of envenomation and hypersensitivity Beyond stings, there are recently described cases of severe allergic reactions to edible jellyfish consumption It has also been reported that jellyfish stings, particularly among surfers, lead to sensitisation of foods containing gamma-glutamic acid, such as fermented soybean , Collectively, these data highlight the antigenic properties within jellyfish nematocysts and tissue, in addition to their venom-derived destructive potential.
Systemic inflammation, including cytokine release syndrome CRS , is a life-threatening immune condition triggered in response to endotoxemia, severe viral infections including influenza , and immunotherapies — Clinical manifestations can include fever, nausea, tachycardia, dyspnea, headache, muscle and joint pain, and in severe cases, neurotoxicity, pulmonary oedema, respiratory failure, and death Certain envenomations can similarly provoke a systemic inflammatory response, most notably is scorpionism Scorpion envenomation is another leading cause of venom-associated morbidity and mortality, affecting more than one million individuals per annum 6.
Additionally, venom-derived toxins induce spontaneous acetylcholine Ach release from peripheral nerves, responsible for the life-threatening cardiac dysfunction seen in severe cases Interestingly, these mediators are also implicated in the box jellyfish pathology, Irukandji syndrome Along with intense acute pain and distress, severe scorpion envenoming grade III stings produces complex pathophysiology in victims Like CRS, symptoms can include respiratory distress, cardiac dysfunction, pulmonary oedema, multiple organ failure, and potentially death, especially among children and the elderly Ion channel modulation is also implicated in the development of pulmonary oedema, a symptom present in many fatal sting cases Research led by Comellas et al.
Beyond neurotoxic effect, the immune network plays a role in significant scorpion envenomations Immune participation in SR is multifactorial, involving direct antigenic activation and indirect stimulation via the neuroendocrine-immune axis In addition to neurotransmitters, stings induce a rapid release of proinflammatory mediators.
Elevated IFN-γ, IL-1β, IL-6, IL-8, IL, and TNF have been detected in the plasma of sting patients and animal models of scorpionism Scorpion venom also provokes hypersensitivity mediators, particularly histamine Blockade of the histamine H1 receptor has shown to be protective in Androctonus australis hector envenomed mice Specifically, pretreatment with hydroxyzine H1 receptor antagonist reduced immune cell infiltrate and oedema in the brain and spinal cord and diminished levels of circulating proinflammatory cytokines Further, scorpion toxins activate components of the complement system, including the generation of anaphylatoxins, which are potent chemotactic proteins As further evidence of immunological involvement, heightened dermal reactions are reported in individuals predisposed to scorpion venom, such as seen in delayed-type hypersensitivity reactions For direct immunological activation, the most extensively studied species is the Brazilian scorpion, T.
serrulatus Whole T. serrulatus venom TsV and select purified toxins are potent stimulators of innate immune cells, including MΦs In vitro assays have revealed that surface receptors TLR2, TLR4, CD14, and CD36 recognise TsV compounds, triggering cellular activation and production of cytokines and lipid mediators Engagement of CD14 and co-receptor TLR4 promotes NF-κB and AP-1 signalling pathways and transcription of potent proinflammatory genes, including IL-1β Consequently, TsV stimulates cytokine release from innate immune cells in a time- and dose-dependent manner, independent from cytotoxic effect In addition, NF-κB signalling regulates cyclooxygenase-2 COX-2 expression and the secretion of eicosanoid, PGE 2 PGE 2 is a lipid mediator with pleiotropic roles in the initiation and resolution of inflammation, particularly inflammatory pain Among its diverse biological functions, PGE 2 activates IL-1β, MCP-1, and IL-6 pathways via prostaglandin EP4 receptor signalling Accordingly, IL-1β and its receptor IL-1R are strongly suppressed by EP4 antagonism IL-1β and IL-1R are potential therapeutic targets for multiple inflammatory diseases, including scorpion envenomation The eicosanoid leukotriene B4 LTB 4 is also upregulated in cell culture and animal plasma following treatment with whole TsV or purified toxins A study by Zoccal et al.
CD14 and TLR4 appear to be critical for TsV-induced cytokine and eicosanoid secretion Critically, LTB 4 synthesis suppresses IL-1β maturation and secretion and the associated animal mortality CD36, therefore, represents a novel therapeutic target for severe scorpion envenomation Mouse models of TsV envenomation produce autonomic dysfunction that is similar to clinically observed symptoms A lethal inoculation of TsV induces sweating, ocular and nasal secretions, lethargy, and convulsions in mice, preceding cardiovascular disturbances and death Observed hyperglycemia and neutrophilia are also consistent with sting patients The neurotransmitters adrenaline and ACh, responsible for sympathetic and parasympathetic symptoms, respectively, are elevated in peripheral blood as well as in cardiac tissue in response to TsV Treatment with atropine, a muscarinic receptor antagonist, but not propranolol, prevented venom-induced cardiovascular alterations, which are a leading cause of death in severe scorpionism Curiously, despite showing systemic elevation of adrenaline, the study did not investigate the effect of an alpha-adrenergic blocking agent, such as prazosin In parallel to excessive ACh, lethal TsV envenomation stimulates the systemic and cardiac secretion of PGE 2 and IL-1β.
Reis et al. have recently proposed IL-1R as a neuro-immune link responsible for innate heart inflammation and TsV-induced heart failure Research by their group demonstrated that TsV co-administered with PGE 2 enhanced IL-1β and ACh release from cardio fibroblasts, an effect which was blocked by an EP receptor antagonist.
In contrast, IL-1R silencing repressed PGE 2 , IL-1β and ACh levels and rescued mice from fatal TsV administration As such, the study determined that PGE 2 amplifies IL-1β release, which upon binding IL-1R potentiates upregulation of PGE 2 and PGE 2 -dependent ACh release post TsV envenomation Existing scorpion sting management comprises specific antiserum and symptomatic treatment, such as pain and low dose anti-inflammatory medications 6.
Grade III stings and stings in children younger than 15 require intensive care 6. Polymorphism within scorpion venom-derived proteins is geographically varied, impeding the manufacture of a standardised antivenom 6.
Variability in toxin immunogenicity further limits the usefulness and cost-effectiveness of antivenom production 6. Accordingly, some experts challenge the use of antiserum therapy due to insufficient neutralising capacity and the additional shock risk associated with poorly purified serum 6 , Recently, success has been reported using novel immune-based therapies in animal models of TsV envenomation.
In , Zoccal et al. Indeed, the therapeutic administration of EP at 0. Lymphocytes and neutrophils in the bronchoalveolar lavage fluid were significantly lower in the treatment group than venom alone.
Accordingly, cAMP concentrations and proinflammatory cytokines IL-1β, IL-6, TNF, and CCL3 were also considerably decreased Although promising, the estimated time and cost of developing a new drug and bringing it to market is 10 — 15 years and hundreds of millions of dollars Conversely, drug repurposing circumvents the requirement for lengthy and expensive preclinical development.
In reviewing the effects of jellyfish venom on the immune system, Tibballs et al. highlighted similarities between the clinical features of Irukandji syndrome and scorpion envenomation While the mechanisms underpinning the pathology of Irukandji syndrome have remained unresolved for decades, they may likewise involve both autonomic and inflammatory pathways.
If so, immune-based therapies may also prove beneficial in severe box jellyfish envenomation and warrant further investigation. While most jellyfish stings do not require medical attention, several species found in tropical waters constitute a public health threat In Australia, the box jellyfish Chironex fleckeri and Carukia barnesi are of particular medical relevance.
A high C. fleckeri venom dose can cause rapid and fatal cardiac arrest In contrast, the smaller box jellyfish, C. barnesi , induces an extremely painful systemic pathology known as Irukandji syndrome IS — barnesi was the first confirmed causative agent of IS after its namesake, Dr Jack Barnes, famously subjected himself, his nine-year-old son, and a local lifeguard to intentional envenoming in Yet, due in part to the elusive nature of this highly venomous jellyfish, research over the years has failed to unravel the mechanisms behind the distinctive syndromic illness , barnesi are small and transparent, with the medusal bell measuring ~20 mm wide Figure 1 , As with other carybdeids, C.
barnesi have a single tentacle per pedalium , Both bell and tentacles are covered with nematocysts, comprising distinct venom composition Figure 1 Image of Carukia barnesi jellyfish. Images showing A close up and B relative size of adult C. barnesi jellyfish. Prof Jamie Seymour pictured.
Photos were taken by A Jamie Seymour JCU, Cairns, Australia and B Rachael Ryan JCU, Cairns, Australia.
A retrospective case study of marine sting presentations to Cairns Base Hospital revealed a wide variation of symptom severity among individuals Of the 39 patients with skin scrapings consistent with C. barnesi nematocysts, some experienced only minor symptoms, while in others, envenomation proved fatal Typically, an IS presentation includes a mild local reaction followed by a characteristic incubation period of five to 60 min before the onset of systemic effects Figure 2 , Pain in the abdomen, chest, lower back, limbs, and joints, is severe, often intractable to opioids and accompanied by extreme distress and agitation , In parallel, the manifestation of tachycardia, hypertension, diaphoresis, dyspnea, and in some instances, priapism may occur , In severe cases, life-threatening complications, such as cardiomyopathy and cardiogenic shock, can arise Due to the significant cardiac dysfunction associated with C.
barnesi envenomation, cardiogenic pulmonary oedema may develop , Tragically, venom-induced intracerebral hemorrhage resulted in the death of two individuals in , Figure 2 Local response to C. barnesi envenomation. Image showing a typical dermal reaction on the arm following a sting from a C.
The red marker indicates the sting site. Photo by Jamie Seymour JCU, Cairns, Australia. barnesi is only one of several causative species, an antivenom is unlikely to be produced Therefore, treatment of severe envenomation is heavily reliant upon opioid-based pain management and symptomatic supportive care, with a mean expected hospital stay of 1.
The clinical manifestations of IS have been attributed to excessive catecholamine release, such as seen in pheochromocytoma, scorpionism, or funnel-web spider envenomation , , Supporting this hypothesis, adrenaline and noradrenaline have been transiently detected in the plasma of C.
barnesi experimentally envenomed piglets Peak catecholamine release was observed 10 min post intravenous IV venom administration, coinciding with the onset of systemic and pulmonary hypertension and tachycardia Plasma catecholamines remained elevated in envenomed animals until 60 min but declined to non-significant levels within 2 h These data suggest the presence of a presynaptic neuronal voltage-gated sodium channel agonist within the venom, as well as the presence of a TTX-insensitive vasoconstrictor , — While a physiological stress response towards IV administration of any toxin may similarly stimulate adrenaline and noradrenaline release, the authors reported the uniqueness of the reaction compared to other box jellyfish venom Research by Ramasamy et al.
barnesi envenomed rats, further supporting the role of endogenous catecholamines in the pathogenesis of IS However, the residual pulse pressure observed in the study suggested the contribution of factors besides catecholamines Furthermore, the result was not reproduced by Winkel et al.
with 0. Unfortunately, as both cardiovascular studies required the use of anesthetised animals, euthanised after 2 h, the critical evaluation of later time points was not possible Yet, in sting victims, symptoms can remain for days, potentially requiring intensive care Regardless, in line with these findings, current clinical guidelines recommend magnesium sulphate MgSO 4 therapy to attenuate pain and suppress excessive catecholamine release in severe IS Its success in doing so has generated divided opinions The results of a randomised trial completed in , and reviewed in , were unable to confirm the ability of MgSO 4 infusion to reduce opioid requirement , Both studies reported varied success from the 39 patients, ultimately showing no significant benefit from MgSO 4 therapy , Akin to scorpion envenomation, the symptoms of IS cannot be wholly attributed to sympathetic hyperstimulation Also akin to scorpionism, generalised IS symptoms resemble those of CRS.
Interestingly, MgSO 4 potently suppresses MNC-mediated cytokine production following TLR stimulation MgSO 4 increases IκBα levels in MNCs, thereby decreasing NF-κB nuclear translocation and its activity Accordingly, the ability of MgSO 4 to inhibit pain in some sting patients could in part be due to a dampened immune response, although this theory has not been investigated.
Presently, neither catecholamines nor inflammatory mediators have been measured in C. barnesi sting patients. Recently, a study by Staedtke et al. This study showed that adrenaline contributes to the positive feed-forward cytokine dysregulation seen in CRS Encouragingly, the blockade of α 1 -adrenergic receptors also expressed on immune cells or the inhibition of tyrosine hydroxylase required for catecholamine biosynthesis by prazosin or metyrosine hindered the self-amplifying proinflammatory loop in vitro and in vivo Specifically, CRS, induced by humanised CD19 CAR-T cells in mice engrafted with a leukemia cell line, caused excessive levels of adrenaline, noradrenaline, and myeloid-derived cytokines IL-6, KC, MCP-1, and TNF in the plasma of animals with high tumour burdens Consequently, higher mortality was observed among these mice.
In contrast, pretreatment with metyrosine or prazosin lowered circulating catecholamines and cytokines, improving survival In mouse peritoneal MΦs, LPS-stimulation induced the release of catecholamines and proinflammatory cytokines, IL-6, KC, MCP-1, and TNF Conversely, reduced MΦ catecholamine production significantly reduced IL-6, KC, MCP-1, and TNF levels Although this model is unrelated to envenomation, it suggests that immune stimuli, such as venom-induced TLR activation, initiates a proinflammatory response that is enhanced by the presence of catecholamines.
The dual stimulatory signals create a positive feed-forward loop, resulting in cytokine amplification. Collectively, these studies suggest a therapeutic potential of prazosin for severe IS, except where contraindicated, and warrant the investigation of plasma cytokines in sting patients.
However, while such research supports the therapeutic inhibition of catecholamines in envenomation, understanding their possible protective function has not been investigated. For example, it is known that catecholamine release promotes alveolar fluid clearance In scorpionism, a rapid catecholamine surge following a dangerous sting is reasoned to increase alveolar fluid reabsorption, protecting the lungs from venom-induced flooding Therefore, the resolution of the surge may decrease the ability of the lungs to clear venom-induced oedema, ultimately progressing toward fatal pulmonary oedema Accordingly, animal models of IS should thoroughly scrutinise both the benefits and limitations of these mediators.
Nevertheless, to date, no C. barnesi immunology-based research has been published. Despite the health burden of human envenomation, venom-immune interactions have been exploited in traditional medicine for centuries More recently, research groups throughout the world have demonstrated the in vitro and in vivo efficacy of whole venom and venom-derived compounds in ameliorating a wide range of autoimmune symptoms , — Presently, the most promising drug leads belong to the class of ion channel modulators.
Ion channels, particularly calcium-activated and voltage-gated potassium channels, are attractive therapeutic targets for autoimmune diseases. Firstly, ion channels, such as the Shaker-related voltage-gated K V 1. Secondly, unique ion channel dimers are differentially expressed in various tissues, including immune cell subsets, permitting cell type and subset-specific blockade , For example, activated effector memory T cells T EM , B cells and MΦs, known mediators in the pathogenesis of various autoimmune diseases, preferentially upregulate K V 1.
In contrast, naïve T n and central memory cells T CM express KCa3. Venom from snakes, spiders, scorpions, cone snails, and sea anemones comprise a diverse range of peptide and small molecule ion channel blockers that exhibit selectivity at picomolar concentrations Blockade of lymphocyte ion channels using venom-derived compounds has therapeutic effects in animal models of rheumatoid arthritis RA , asthma, multiple sclerosis MS , delayed-type hypersensitivity, and allograft rejection , , — Notably, a selective peptide blocker, Stichodactyla helianthus toxin ShK , from sea anemone venom, and anuroctoxin, a peptidyl toxin isolated from Buthus sindicus scorpion venom, have been shown to specifically target K V 1.
Structural studies centred on the selectivity of peptide ion channel blockers have revealed that specificity is due to single amino acid effects rather than en bloc backbone structure Thus, venom-derived peptides may act as promising drug scaffolds, notably because disulphide bonds encode robust biological stability This has important implications for drug development, as synthetic manipulation may improve drug activity or remove toxicity from the natural peptide blueprint.
For example, ShK L5 , a synthetic analog of ShK, contains an N-terminal L-phosphotyrosine extension and shows higher selectivity than the native peptide for K V 1. Aside from ion channel blockade, venom-derived components have demonstrated potent in vitro and in vivo anti-inflammatory activity through the cholinergic anti-inflammatory pathway via an alpha7 nicotinic acetylcholine receptor antagonist Venom-derived peptides, such as the α-neurotoxin from the Thailand cobra, are potent nicotinic receptor antagonists In a rodent RA model, Cobratoxin-treatment reduced expression of the pro-inflammatory cytokines IL-1β, IL-2, and TNFα, resulting in decreased paw sensitivity and joint destruction Other neurotoxins, such as the principal toxin NTX from Naja atra venom NNAV , have shown therapeutic effects in animal models of adjunctive arthritis, RA, Systemic Lupus Erythematosus SLE , and nephropathy , Additionally, NTX-treatment prolonged skin allograft survival in rats and inhibited cell-mediated immune responses in a dose-dependent manner through decreased Th1-type cytokines IL-2 and IFN-γ.
Although low NTX concentrations were cytotoxic, heat-treatment reduced NTX toxicity without reducing its immunosuppressive activity Venom from the honey bee has been used for centuries in traditional medicine to treat chronic inflammatory diseases due to its reported anti-inflammatory activity Investigations into the mechanism of action of honey bee venom and its major components, melittin and phospholipase A2, have confirmed a protective effect in animal models of asthma and RA , The polarisation of T cells towards a Th2 phenotype is associated with allergies and chronic inflammatory diseases An essential driving factor in lineage determination is cytokine expression.
It has been shown that melittin inhibits LPS-induced inflammation by binding to the C- terminus of the NF-κB p50 subunit, thus preventing translocation into the nucleus and transcription of pro-inflammatory cytokines, including TNF — Conversely, PLA 2 , an enzyme found within the venom of multiple species, including the western honey bee, can hydrolyze membrane phospholipids and induce Th2 cytokine responses through the activation of ST2, a component of the IL receptor on innate immune cells Other known venom immune modulators include tick salivary protein Salp 15 from Ixodes scapularis and spermine.
Salp15 binds the CD4 co-receptor, MHC-II, inhibiting TCR ligation and T cell activation by misaligning CD4 with the TCR complex Spermine, an acylpolyamine found in snake and spider venom, suppresses mitogen-induced activation and proliferation of PBMCs by inhibiting LAF-1 protein expression, involved in RNA remodelling Collectively, these studies highlight the potential of venom-derived molecules to modulate immune cells as unmodified venom-derived compounds or as scaffolds for drug development.
Venom-derived compounds induce immune suppression using diverse modes of action. Thus, screening venom for its immunosuppressive and immune-activating potential may result in new immunomodulatory drugs and the discovery of new biological pathways.
Extensive research focuses on the degree to which immune responses themselves contribute to the severity of envenomation Figure 3. Perhaps the most significant cause of division lies in the difficulty of distinguishing the actual venom-induced symptoms from immune-induced pathology. The classic inflammation markers heat, pain, redness, swelling, and loss of function are typical biological responses to envenomation across many species.
Therefore, determining which symptoms are treatable using immunological approaches requires further research. Figure 3 Immunological responses to envenomation. Created with BioRender. Firstly, continued research could inform improved treatment strategies for fatal bites and stings.
Writing—original draft preparation, RR. Writing—review and editing, RR, JM, MI, JS, AL, and JL. Supervision, JM, MI, and JL. Funding acquisition, JM. All authors contributed to the article and approved the submitted version.
RR was supported by an Australian Government Research Training Program stipend. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
I would very much like to thank Dr. Paul Giacomin for critical reading of the original draft manuscript. Jouiaei M, Yanagihara AA, Madio B, Nevalainen TJ, Alewood PF, Fry BG.
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In: Rich RR, Fleisher TA, Shearer WT, Schroeder HW, Frew AJ, Weyand CM, editors. Clinical Immunology. They typically live in communal areas with many snakes sharing the same den. Like the rattlesnake, many of copperheads will shake their tail before a strike, but without the rattle, it is just for show.
However, some may release a very strong musk scent when agitated. During the spring and fall copperheads spend the day out hunting and basking in the sun; however, in the middle of summer, they often become nocturnal, making a strike harder to avoid.
Copperheads are more common and bite more people than any other species of snake. Thankfully, they have the mildest venom of all North American venomous snakes. But their bites still require emergency medical attention.
Researchers have found that while many snakes — both venomous and nonvenomous — give a warning and prefer to retreat, copperheads prefer to stand their ground and strike.
These snakes are very territorial, often returning to the same den year after year. Copperheads have a lifespan of around 18 years. So if you reside in an area where this venomous snake lives, keep your yard, underneath your deck, and garage free from clutter, wood piles, and the like to help prevent them from moving in and bringing friends.
They are most commonly found in the southeastern United States, as far north as Virginia, and as far west as Texas. They like fresh water ponds, lakes, streams, swamps and marshes; however, they also dwell in drainage ditches.
Water moccasins can be aggressive, but they rarely bite humans. On land, they will coil and open their mouths wide, exposing the white cottony lining. This is why they are also known as cottonmouths. They range in length from 50 to 55 inches long and have a dark tan, brown or nearly black skin with cross bands of black or brown.
These venomous snakes use their time on land to soak up the warmth of the sun. They are often found basking in the sunshine lying on stones, logs or even hanging from tree branches.
While in the water, they swim close to the top, with their heads out. During the summer months, they hunt after dark. Their venom can be deadly; the hemotoxins can prevent the blood from clotting, resulting in hemorrhaging throughout the circulatory system. If you are bitten, you must seek emergency medical attention immediately.
There are a number of nonvenomous snakes that can be confused with the water moccasin; if you can safely take a photo, do so to show to your medical team.
Coral Snakes. Small and colorful, coral snakes have the strongest venom of any snake in North America. They are second only to the black mamba, the snake with the most deadly venom on the planet. These small snakes are 18 inches to 20 inches long and are very slender, almost pencil-like.
There are two types of coral snakes: the eastern coral snake and the western coral snake. Both species are reclusive, love marshy or wooded areas, or hiding under leaves, debris or in woodpiles. The eastern type is primarily found in the Southeast, from North Carolina to Florida, and west to parts of Texas.
The western coral snake is mainly found in southern Arizona and the southwestern most tip of New Mexico. Both types of coral snakes have bright colored bands of yellow, red and sometimes black. Because there are several other non-venomous snakes in North America with similar colors, including the scarlet kingsnake and the milk snake, err on the side of caution if a snake bites you.
The venom of a coral snake is a neurotoxin, which can cause paralysis, respiratory failure and cardiac failure. Because these snakes are so brightly colored and are generally small and thin, they tend to attract young children.
It is imperative that you teach your child to never handle a snake without adult supervision. Most snakes in North America, are not poisonous. However, the four snakes mentioned above do have venom that can cause severe adverse health conditions — and even death — if not treated properly and promptly.
Whenever you are bitten by a snake, it is recommended that you seek emergency medical attention immediately; even nonvenomous snake bites can cause problems. Bacteria and viruses in snake mouths can lead to nasty infections and permanent scarring.
After striking, a snake may stick around and try to strike again, or it may slither off. It is virtually impossible, particularly in the heat of the moment, to accurately identify a snake. There are literally thousands of species, many of which appear similar to an untrained eye.
That said, if you live in an area or are traveling to one that is known for venomous snakes, it is wise to learn the basic physical characteristics. However, remember that snakes are nearly universally feared and when you encounter one, your adrenalin levels will increase, making it even more challenging to remain calm while trying to identify it.
Always err on the side of caution after a snake bite. If possible, and without risking another bite, take a photo to show to the emergency responders. A snake can strike quickly, biting with extreme force, and then slither away just as fast.
Many snake bites occur when people are working in their yards or gardens, when children are playing by a creek or in a wooded area, and when taking part in recreational activities. All of this to say that a snake can strike virtually any time, anywhere.
When you are first struck, you may notice two puncture wounds and experience quite shocking pain. Some describe the pain as being stung by a thousand bees, all in the same spot.
Swelling and redness appear quite quickly with the most bites; however, when bitten by a coral snake, symptoms, including pain, may not appear for 12—18 hours.
If a venomous snake bites you, the bite will likely introduce venom into your system. Bites by dangerous snakes are greatly feared, and effective symptomatic diagnosis and treatment are poorly documented.
Due to complex pathogenesis and interspecific variability of venom components, the mechanisms and pathophysiological changes are still not completely elucidated [ 8 8. Gren ECK, Kitano ES, Andrade-Silva D, Iwai LK, Reis MS, Menezes MC, et al.
Comparative analysis of the high molecular mass subproteomes of eight Bothrops snake venoms. Comp Biochem Physiol Part D. Fatima L, Fatah C. Pathophysiological and Pharmacological Effects of Snake Venom Components: Molecular Targets.
J Clin Toxicol. Symptoms induced by snake venoms are multi-factorial and complex, which severely restricts therapeutic intervention after the snake bite [ 10 Mehta SR, Sashindran VK.
Clinical Features And Management Of Snake Bite. Med J Armed Forces India. Moreover, differing components of different venoms make it difficult to generate sufficient diagnostic markers for clinical practice in both Western medicine and Chinese medicine [ 11 Tan CH, Wong KY, Chong HP, Tan NH, Tan KY.
Proteomic insights into short neurotoxin-driven, highly neurotoxic venom of Philippine cobra Naja philippinensis and toxicity correlation of cobra envenomation in Asia. J Proteom.
Fox JW, Serrano SM. Timeline of key events in snake venom metalloproteinase research. J Proteomics. Quantitative proteomic analysis of multiple venoms revealed that the venom proteome composition includes non-enzymatic and enzymatic proteins, such as three finger toxins 3FTx and phospholipase A 2 PLA 2 [ 13 Patra A, Chanda A, Mukherjee AK.
Quantitative proteomic analysis of venom from Southern India common krait Bungarus caeruleus and identification of poorly immunogenic toxins by immune-profiling against commercial antivenom.
Expert Rev Proteomics. Maduwage K, O'Leary MA, Isbister GK. Diagnosis of snake envenomation using a simple phospholipase A2 assay.
Sci Rep. In the absence of sufficient diagnostic markers, the clinical identification of a venomous snake is mainly based on the patient's wound tooth distance, tooth marks, local symptoms, and consultations. Traditional Chinese medicine TCM classifies venomous snakes on the basis of toxicity symptoms into three types: wind toxin, fire toxin, and wind-fire toxin [ 16 Deng T, Ergil K.
Practical diagnosis in traditional Chinese medicine. Edinburgh ; New York: Churchill Livingstone, The fire toxin is similar to hemorrhagic venom. Clinical manifestations of snakebites by fire toxin snakes include local pain, immediately swelling, water or blood blisters around the bite on the affected limb, skin ecchymosis, ulceration, necrosis, and general hemorrhage.
The wind toxin can be regarded as part of neurotoxic venoms, generating mild pain, minor blood loss, numbness, somnolence, difficulty in opening the mouth, confusion, respiratory distress, blurred vision, etc. The wind-fire toxin is similar to mixed venom, which presents a combination of neurotoxic and hemorrhagic symptoms.
The wind-fire toxin provokes respiratory distress, drooping eyelids, blurred vision, muscle weakness, and paralysis of muscles in the extremities. In our TCM clinical work, fire toxin and wind-fire toxin are particularly prevalent, so that the predominant venomous snake species in China is deserving prior attention, including Agkistrodon acutus and Trimeresurus stejnegeri snakebite fire toxin , and viper and cobra snakebite wind-fire toxin.
In TCM, the same treatment strategy is implemented for the same snakebite syndrome, even in different patients that were bitten by different snakes. At present, the identification of the snakebite syndrome in TCM relies on expert judgment and knowledge, such as diagnosis by exclusion, and speculative and empirical judgments.
The emergency treatment of snakebites is based primarily on clinical practice guidelines, some of which lack scientific study. Furthermore, our understanding of the relationship between Compound Chinese Medicine and the effects of multiple components is still in its infancy [ 17 Sun K, Fan J, Han J.
Acta Pharm Sin B. Rapid and accurate identification of the snakebite etiology is also an important prerequisite for timely and effective treatment in TCM. It is urgent to correlate the pharmacological properties to clinical manifestations for a better understanding of wind-fire toxin and fire toxin.
Investigation and analysis of the protein composition of serum from snakebite patients for diagnostic biomarkers will contribute to the clinical practice. This study aims at solving diagnostic markers and investigating pathways of snake venom-induced toxicity from serum proteins of snakebite patients with four predominant venomous snake species in China.
The serum protein profiles reflected the functional categorization. The differential abundance proteins DAPs supplied discovery biomarkers to distinguish snakebite by species.
The network of interacting proteins and drug compounds contribute to understanding the complex pathogenesis of venom-induced toxicity. Serum proteome analysis could contribute to TCM diagnostic markers of snakebite syndromes, providing clinicians with a clear distinction between the wind-fire toxin and fire toxin, as well as early clinical identification of snake species.
The experimental procedures were approved by the Ethics Committee of the Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine n. Informed consent forms were obtained from all recruited participants in the surgical department of the Traditional Chinese Medicine of the Affiliated Hospital.
All identified snakebite patients were reviewed for eligibility to inclusion and exclusion criteria. Patients were excluded for the following reasons:. Clinicians identified the pattern of fire toxin and wind-fire toxin according to the standards of TCM, including clinical diagnostic experience and patient medical records.
From the serum samples available, we selected four cases bitten by vipers B and three cases bitten by cobras C as the wind-fire toxin group; four cases bitten by Agkistrodon acutus D and three cases bitten by Trimeresurus stejnegeri E as the fire toxin group; and fifteen healthy volunteers as the control group.
The serum samples from 5 healthy controls were pooled together, so that the individuals in the control group were divided into three experimental duplicates, named A1, A2, and A3. The wind-fire toxin group was coded B1, B2, B3, B4, C1, C2, and C3; while the fire toxin group was coded D1, D2, D3, D4, E1, E2, and E3.
All of the serum samples were processed as described previously [ 18 Tu C, Rudnick PA, Martinez MY, Cheek KL, Stein SE, Slebos RJC, et al. Depletion of abundant plasma proteins and limitations of plasma proteomics. J Proteome Res. Briefly, high abundance proteins were first depleted by multiple affinity removal according to the Agilent protein purification protocol [ 19 Agilent Technologies.
Agilent Human 14 Multiple Affinity Removal System Columns for the Fractionation of High-Abundant Proteins from Human Proteomic Samples. Agilent Technologies, Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. The gel was stained with Coomassie blue, then re-suspended with dithiothreitol and digested with the sequence grade modified Trypsin Promega [ 21 Candiano G, Bruschi M, Musante L, Santucci L, Ghiggeri GM, Carnemolla B, et al.
Blue silver: a very sensitive colloidal Coomassie G staining for proteome analysis. The digested proteins were passed through a C18 cartridge to be desalted. Finally, the dried peptides were resuspended in 40µL of 0.
Then 2 µL peptide injection volume was loaded via a direct inject column configuration onto an Easy-nano LC instrument Thermo Fisher Scientific coupled online to a Q-Exactive Plus Thermo Fisher Scientific.
The resuspended peptides were first separated on Thermo Acclaim PepMap RSLC nanoViper C18 column 50 µm×15 cm using solvent A 0. Bhosale SD, Moulder R, Kouvonen P, Lahesmaa R, Goodlett DR. Mass spectrometry-based serum proteomics for biomarker discovery and validation.
Methods Mol Biol. Cox J, Neuhauser N, Michalski A, Scheltema RA, Olsen JV, Mann M. Andromeda: a peptide search engine integrated into the MaxQuant environment. Cox J, Mann M.
MaxQuant enables high peptide identification rates, individualized p. Nat Biotechnol. The UniProt human isoform protein database was downloaded in August , and contained , entries at this time.
For the searches, the first search and mass tolerance of 20 ppm and two max missed cleavages were allowed. Fixed modification was set to carbamidomethyl C , while variable modifications were oxidation M and acetylation protein N-terminal.
A false discovery rate FDR of 0. Matching and alignment time window in match between run options were set to 0. Furthermore, only unique proteins identified with more than unique razor peptides were considered [ 24 Finally, a label-free quantification LFQ approach was carried out for normalization of protein intensities across runs with a minimum of one ratio count [ 23 The LFQ measuring intensities were assigned to the relative abundance for each protein and used in the subsequent statistical analysis.
At least two non-null values of data in each group were required for calculation of fold change and p -values. In addition, we took into account a comparison of all missing values in one group with expression values in another group.
Proteins that were expressed in one group and not expressed in another group were considered to have a significant difference between the two groups. For the functional annotation of identified proteins, the Gene Ontology GO and UniProt databases were used.
GO enrichment analyses of differentially expressed proteins were performed with Fisher's exact test. Metsalu T, Vilo J. ClustVis: a web tool for visualizing clustering of multivariate data using Principal Component Analysis and heatmap.
Nucleic Acids Res. Protein-protein interaction PPI networks were conducted using the STITCH 5 database [ 26 Szklarczyk D, Santos A, von Mering C, Jensen LJ, Bork P, Kuhn M.
STITCH 5: augmenting protein-chemical interaction networks with tissue and affinity data. Szklarczyk D, Franceschini A, Wyder S, Forslund K, Heller D, Huerta-Cepas J, et al. STRING v protein-protein interaction networks, integrated over the tree of life.
Label-free quantitative proteomics of serum samples was performed on samples from 14 subjects with four types of snakebites and 15 healthy controls. Of the 14 patients, 7 were diagnosed by TCM to have wind-fire toxin including 4 snakebites by vipers B and 3 snakebites by cobras C , and 7 were diagnosed by TCM to have fire toxin including 4 snakebites by Agkistrodon acutus D and 3 snakebites by Trimeresurus stejnegeri E.
The fifteen healthy controls were pooled into three samples as described in the methods. Peptide identification reported using MaxQuant analysis.
Protein quantification with LFQ intensity profiles obtained after normalization by the MaxLFQ algorithm. The representative proteins for each group were comprised of , , and proteins in healthy control, wind-fire toxin, and fire toxin groups, respectively.
When we filtered the results to satisfy the condition of getting at least two non-null intensities in replicates, we identified unique proteins between patients with snakebites and healthy control. We initially examined whether the serum proteins could discriminate between the two snakebite groups and the healthy control group.
We identified proteins in both patient groups, constituting Based on the serum protein expression profiles, the three groups were differentiable by principal component analysis PCA Figure 1B. We observed that the healthy control group was clearly separated from the bulk of snakebite patients at the first principal component PC1 , accounting for
The Dftoxification for Snake venom detoxification methods Control and Prevention Natural remedies for diabetes indicates that venomous snakes bite somewhere between deyoxification, and 8, people in the United States each year, Detoification about five die. In the U. Rattlesnake: The largest venomous snake in the U. There are 29 species of the rattlesnake, found far and wide across the country. From grassy meadows in the Northeast to swamps in the Southeast — even in the mountains, as they can thrive at up to 11, feet in elevation. However, they are most prevalent in the Southwestern part of the U. This Snake venom detoxification methods aimed to evaluate the clinical therapeutic efficacy deoxification anti-snake venom serum blockade in treating local tissue necrosis caused methoss Chinese Nutrient absorption in the stomach Naja merhods bites. The experimental group received regular as well as anti-snake detoxificstion serum blocking treatment, whereas regular treatment plus chymotrypsin blocking therapy was given to the control group. The necrotic volumes around snake wounds in these groups were detected on the first, third and seventh days. On the third day of treatment, some local tissues in the wounds were randomly selected for pathological biopsy, and the necrosis volume of the local tissue was observed. Furthermore, the amount of time required for wound healing was recorded.
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