melitten and Hypersensitivity

Melittin is the main effective component of bee venom and has extensive biological functions; however, serious side effects have restricted its clinical application. Preclinical and clinical studies showed that the main adverse events were allergic reaction and pain at the administration site. To decrease the toxicity, we prepared melittin nano-liposomes by encapsulating melittin with poloxamer 188 and explored the inhibitory activities on liver cancer together with biological safety. Here, we showed that melittin nano-liposomes significantly inhibited the survival of hepatocellular carcinoma (HCC) cells in vitro and prominently suppressed the growth of subcutaneous and orthotopic HCC transplantation tumors in vivo. It was important that it induced less inflammation and allergy in mice compared with melittin. Overall, melittin nano-liposomes would have a better application in HCC therapy due to its significant anti-tumor activity and better biological safety.

Topics: Animals; Bees; Capsules; Carcinoma, Hepatocellular; Heterografts; Humans; Hypersensitivity; Inflammation; Liposomes; Liver Neoplasms; Melitten; Mice; Nanoparticles; Poloxamer

Topics: Adult; Allergens; Animals; Bee Venoms; Bees; Biomarkers; Desensitization, Immunologic; Disease Progression; Female; Humans; Hyaluronoglucosaminidase; Hypersensitivity; Immunization; Immunoglobulin E; Insect Proteins; Male; Melitten; Middle Aged; Phospholipases A; Young Adult

Venoms consist of a complex mixture of toxic components that are used by a variety of animal species for defense and predation. Envenomation of mammalian species leads to an acute inflammatory response and can lead to the development of IgE-dependent venom allergy. However, the mechanisms by which the innate immune system detects envenomation and initiates inflammatory and allergic responses to venoms remain largely unknown. Here we show that bee venom is detected by the NOD-like receptor family, pyrin domain-containing 3 inflammasome and can trigger activation of caspase-1 and the subsequent processing and unconventional secretion of the leaderless proinflammatory cytokine IL-1β in macrophages. Whereas activation of the inflammasome by bee venom induces a caspase-1-dependent inflammatory response, characterized by recruitment of neutrophils to the site or envenomation, the inflammasome is dispensable for the allergic response to bee venom. Finally, we find that caspase-1-deficient mice are more susceptible to the noxious effects of bee and snake venoms, suggesting that a caspase-1-dependent immune response can protect against the damaging effects of envenomation.

Topics: Animals; Apoptosis Regulatory Proteins; Blotting, Western; Bone Marrow Cells; CARD Signaling Adaptor Proteins; Carrier Proteins; Caspase 1; Cell Line, Tumor; Cells, Cultured; Crotalid Venoms; Cytoskeletal Proteins; Enzyme Activation; Hypersensitivity; Immunoglobulin E; Inflammasomes; Inflammation; Interleukin-1beta; Macrophages; Mast Cells; Melitten; Mice; Mice, Inbred C57BL; Mice, Knockout; Neutrophils; NLR Family, Pyrin Domain-Containing 3 Protein; Receptors, Interleukin-1; Venoms

Treatment with aqueous and aluminum hydroxide (Al[OH](3))-adsorbed purified honeybee (Apis mellifera) venom (HBV) preparations can reduce the incidence of side effects associated with venom immunotherapy.. The aim of the present study was to assess these purified HBV immunotherapy preparations in situ.. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was used to visualize the distribution of HBV components. The preparations were administered on the back legs of naive Wistar rats. The rats were killed, and cryosectioned tissue sections were subjected to hematoxylin and eosin staining and MALDI-MSI analyses.. Low-density maps of tissue distribution of HBV peptides, such as secapin, mast cell degranulating peptide, and melittin (Api m 4) were detected in the tissue after administration of HBV immunotherapy preparations. In addition, release of biogenic amines, cytokines, and leukotrienes was observed, and the distribution of HBV allergens, such as Api m 1 and Api m 2, was shown. At the 24-hour time point, the major HBV allergen Api m 1 was still detected at the site of Al(OH)(3)-adsorbed HVB injection, whereas in the case of aqueous HBV preparation, all the allergens, as well as most of the biogenic amines, were cleared at the 24-hour time point.. The present study shows that the majority of low-molecular-weight HBV components are rapidly removed from the site of venom immunotherapy administration. Furthermore, Al(OH)(3)-adsorbed HBV preparation demonstrated a depot effect, prolonging the availability of bee venom allergens at the site of administration.

Topics: Allergens; Aluminum Hydroxide; Animals; Antigens, Plant; Bee Venoms; Bees; Biogenic Amines; Cryoultramicrotomy; Desensitization, Immunologic; Humans; Hyaluronoglucosaminidase; Hypersensitivity; Insect Proteins; Lasers; Melitten; Peptides; Phospholipases A; Rats; Rats, Wistar; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Water

A recent report from our laboratory shows that subcutaneous (s.c.) injection of melittin could induce persistent spontaneous nociception (PSN) and primary thermal or mechanical hyperalgesia. However, the exact peripheral mechanisms underlying melittin-induced multiple pain-related behaviors remain unclear. In this study, behavioral tests combined with pharmacological manipulations were used to explore potential roles of local P2X and P2Y receptors in melittin-induced inflammatory pain and hyperalgesia. Post-treatment of the primary injury site with s.c. injection of A-317491 (a potent P2X(3)/P2X(2/3) receptor antagonist) and Reactive Blue 2 (a potent P2Y receptor antagonist) could significantly suppress the development of melittin-evoked PSN and hypersensitivity (thermal and mechanical). Our control experiments demonstrated that local administration of either antagonist into the contralateral hindpaw produced no significant effect on any kind of pain-associated behaviors. Taken together, these data indicate that activation of P2X and P2Y receptors might be essential to the maintenance of melittin-induced primary thermal and mechanical hyperalgesia as well as on-going pain.

Topics: Animals; Hypersensitivity; Male; Melitten; Nociceptors; Pain; Phenols; Polycyclic Compounds; Rats; Rats, Sprague-Dawley; Receptors, Purinergic P2; Receptors, Purinergic P2X

Melittin, a bee-venom peptide of 26 amino acids, induces IgE and IgG responses in man and animals. The antibody response was shown previously to be specific primarily for the C-terminal 6 residues and its T cell epitope in H-2d restricted mice was shown to be in residue 11-19 of melittin. To study the relationship of peptide structure and immunogenicity in mice, we have prepared a series of melittin analogs varied in length and composition at the C-terminus. Immunogenicity of the analogs for IgG and IgE responses was found to correlate with two factors: a peptide length of more than 24 residues and the presence of a hydrophilic C-terminal region preferably with two to four cationic groups. These factors result in the ability of peptide to bind to cell membranes. Analogs that possess these features are good immunogens whereas those lacking any of these features are weak immunogens.

Topics: Amino Acid Sequence; Animals; Antigen-Antibody Reactions; Epitopes; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Melitten; Mice; Mice, Inbred BALB C; Mice, Inbred DBA; Molecular Sequence Data; T-Lymphocytes

Previous work has demonstrated a close association between certain histocompatibility antigens and the gene that controls the IgE response to certain ragweed allergens. For example, there is a 90% association between IgE production to the short ragweed allergen, Amb a V, and an HLA class II allele. To assess whether these HLA linkages are specific for ragweed, we have investigated the association between HLA antigens and the capacity of individuals to mount a specific IgE response to melittin in patients with bee-venom allergy. Twenty-two subjects with bee-venom sensitivity, 22 healthy beekeepers without bee-venom allergy, and a normal population of 149 unselected individuals were studied. With serologic tissue typing and restriction fragment length polymorphism analysis, we have demonstrated a significant decrease in the HLA-DR4 and DQw3 alleles in subjects who are allergic to melittin compared to the control populations. There was also a negative association between the presence of HLA-DR4 and DQw3 alleles with the capacity of the individuals to mount an IgE response to phospholipase A2 (PLA2). The bee-venom sensitive subjects had a slightly lower titer of anti-PLA2 IgG when these subjects were compared to the bee-venom insensitive beekeepers. These results support the view that either HLA-DR or HLA-DQ has a protective role in controlling the IgE immune response. Lack of an IgE response to melittin or PLA2 is unlikely to be due to a failure to recognize allergen.

Topics: Adolescent; Adult; Aged; Bee Venoms; Female; Histocompatibility Testing; HLA Antigens; HLA-D Antigens; Humans; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Male; Melitten; Middle Aged; Phenotype; Phospholipases A; Phospholipases A2; Polymorphism, Restriction Fragment Length

The immunological response to individual bee-venom allergens was studied in blood samples collected at frequent intervals from four bee-venom allergic patients who had suffered systemic allergic reactions to injections of bee venom during immunotherapy. All had high IgE antibody levels, at the upper end of the range found in bee-sting allergic patients, and all had antibodies to the minor allergens at the time of the reactions. These did not, however, provide a simple explanation for the reactions that occurred. We were able to observe two interesting phenomena--in one patient IgE antibodies to the individual venom antigens appeared to be 'switched off' sequentially. In another, IgE antibodies to hyaluronidase rose substantially after 4 years of therapy. We believe that these results provide evidence to support the view that the regulation of IgE antibodies is controlled by mechanisms that are both isotype- and antigen-specific.

Topics: Acid Phosphatase; Adolescent; Adult; Bee Venoms; Desensitization, Immunologic; Female; Humans; Hyaluronoglucosaminidase; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Male; Melitten; Phospholipases A; Radioallergosorbent Test

IgE antibodies to purified proteins and peptides from honeybee venom have been measured by the RAST. Trace amounts (less than 0.1%) of the major venom protein phospholipase A2 (PLA2) grossly distorted the measurement of IgE antibody to the other venom proteins, acid phosphatase (Acid P) and hyaluronidase (HYAL), and overemphasized their importance. Reduction of antigen coupled to the cellulose paper discs, which were used in the assay, diluted out the contaminating PLA2 without apparent loss in sensitivity. The reduction of disc-bound antigen increased the competition between IgE and IgG antibodies but did not affect measurement of IgE antibodies in sera taken from 35 untreated patients who had a history of general allergic reactions to bee stings. In 54% of sera from bee venom--allergic patients, the greatest IgE antibody response was to PLA2. In all, IgE antibodies to PLA2 were present in 91% of these sera. IgE antibodies to Acid P, HYAL, or melittin were present in 60%, 51%, and 31% of sera, respectively, and accounted for the highest level of binding in 17%, 17%, and 6% of these. Only 6% of sera were positive for whole venom but negative for the isolated antigens. A low level of IgE antibody was found to peptide 401 in 6% of sera. No IgE antibodies were found to apamin. While confirming the central role played by PLA2 in bee sting allergy, these results show that other venom components are also important in some patients.

Topics: Acid Phosphatase; Antibodies; Antibodies, Anti-Idiotypic; Antibody Specificity; Apamin; Bee Venoms; Bees; Humans; Hyaluronoglucosaminidase; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Insect Bites and Stings; Melitten; Phospholipases A; Phospholipases A2; Radioallergosorbent Test

Topics: Animals; Ant Venoms; Apamin; Bee Venoms; Female; Humans; Hymenoptera; Hypersensitivity; Insect Bites and Stings; Melitten; Venoms; Wasps

The IgE and IgG antibody responses to four allergen fractions of honey bee venom were studied in 13 sera from patients who experienced systemic reactions from bee stings and eight from those who experienced large locals. Both groups were similar in allergen response. IgE and IgG levels and ratios. A significant number of patients were found to have experienced both types of reactions from bee stings. It is proposed that IgE antibody positive systemic and large local reactions are both IgE mediated allergic reactions. The determining factors for reaction type include venom dose and sting site.

Topics: Adolescent; Adult; Bee Venoms; Child; Female; Humans; Hyaluronoglucosaminidase; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Male; Melitten; Middle Aged; Phospholipases; Radioallergosorbent Test

Topics: Allergens; Animals; Bees; Histamine Release; Humans; Hyaluronoglucosaminidase; Hypersensitivity; Immunity, Maternally-Acquired; Leukocytes; Melitten; Molecular Weight; Phospholipases; Precipitin Tests; Venoms