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Honeybee venom (apitoxin), composed of bioactive components such as melittin, apamin, and phospholipase A2, has been used in traditional medicine for centuries. Modern research explores its potential therapeutic applications for various diseases. Clinical studies and preclinical research have demonstrated its efficacy in treating conditions like rheumatoid arthritis, osteoarthritis, neurological disorders, and cancer. Despite its therapeutic potential, bee venom carries risks, including allergic reactions. Advances in synthetic venom production offer promising alternatives for broader and safer clinical applications. While more large-scale clinical trials are needed to confirm its efficacy and safety, honeybee venom remains a compelling candidate for novel treatments across a wide range of diseases. [ABSTRACT FROM AUTHOR]

OBJECTIVE Salmonella Typhimurium is a significant zoonotic concern for human food poisoning and a substantial economic burden in the swine industry. We previously reported that nasally delivered chitosan-coated poly(lactide-co-glycolide) (PLGA) encapsulating honeybee venom (CP-HBV) could enhance CD4+ T helper 1 (Th1)-related immune responses in healthy pigs. Building upon these findings, the current study aimed to investigate the protective immune enhancement by nasally delivered CP-HBV in pigs challenged with S Typhimurium. ANIMALS 36 healthy, 4-week-old, female, Landrace X Yorkshire X Duroc pigs. METHODS 36 pigs were allocated into 3 groups: CP-HBV (n = 16), control (n = 16), and healthy baseline control (n = 4). CP-HBV and control groups were challenged with S Typhimurium 7 days post-treatment. Pigs from the healthy control group were sacrificed at 0 days postinfection (DPI), and 4 pigs from each of the control and CP-HBV groups were sacrificed at 1, 2, 4, and 7 DPI. Salmonella shedding, immune cell frequencies, cytokines, and transcriptional factor expression levels were measured. RESULTS The CP-HBV group exhibited lower bacterial shedding and an enhanced Th1-related immune response characterized by an upregulation of CD4+ T cells and CD4+ Interferon-γ+ T cells, accompanied by increased expression of Th1-related cytokines and reduced expression of regulatory T cells and immunosuppressive cytokines compared to the control group. CLINICAL RELEVANCE CP-HBV is a promising strategy for controlling Salmonella infections in pigs and improving public health. [ABSTRACT FROM AUTHOR]

Honeybee venom is one of the natural substances produced by bees (Apis mellifera). Their venom gland produces venom which plays a defensive role. In this study a concentration of macro and trace elements (Ag, Al, As, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Li, Mg, Mn, Mo, Na, Ni, Sb, Se, Sr, Pb and Zn) in foragers′ and honeybees′ venom was analysed by axial inductively coupled plasma optical emission spectrometry (ICP OES) with good validation parameters to differentiate the element accumulation ability in honeybee venom. Cumulative ability for some elements (As, Al, Ba, Cr, Li, Mo, Pb, and Zn) in bee venom was clearly demonstrated. Oppositely, levels of macro elements (Ca, K, Mg and Na) in venom were several times lower compared to the levels detected in foragers. Moreover, PCA analysis of bee samples showed that Cr was associated with locality Košice, and Cd with locality Krompachy; both have rich industrial history. Since some of analysed elements are potentially toxic for humans, a risk assessment for bee-stung scenario was also calculated. A new way of exposure to potentially toxic elements via honeybee stung was showed in this study. Non-carcinogenic risk assessment for humans to selected toxic elements (As, Cd, Cr, Ni, and Pb) demonstrated acceptable risk and moreover the same we may conclude for potential carcinogenic risk for beekeepers exposed to As, Cd, Ni, and Pb via venom over their whole life.

Honeybee venom (HBV) is a product of natural origin with an extremely complex composition. It exhibits several beneficial pharmacological properties, for example anti-inflammatory, antimicrobial, antidiabetic, anti-arthritic, and anti-tumour activities. On the other hand, HBV contains highly allergenic components, including melittin, which makes up more than 50% of its dry weight. Despite many studies conducted, HBV's components and mechanism of action are not yet fully known and understood. Therefore, to enhance the safety of its therapeutical applications, it is essential to establish HBV's composition and biological properties. Many analytical techniques and methods have been reported as potentially useful for the determination of new HBV constituents. Studies focus mainly on the analyses of the HBV proteome and metabolome. Also, properties of its components when applied to the skin, as well as its allergenicity, are being studied. Currently, the most commonly used methods for HBV analysis include high-performance liquid chromatography and mass spectrometry. Choosing the correct and most applicable method for HBV analysis is of great importance in order to obtain in-depth results crucial for the understanding of its mechanism of biological action. The diversity of chemical compounds in HBV also entails an appropriate sample preparation at the pre-analytical stage. In order to summarise the latest knowledge on techniques for the analysis of complex natural products, on the example of HBV, this review paper discusses the modern analytical tools of HBV characterisation, focusing on metabolomic and proteomic studies, topical applications, investigation of venom allergies, and the prospects for future research.

Apitherapy (using bee products) has gained broad recognition in cancer therapeutics globally. Honeybee venom has a broad range of biological potential, and its utilization is rapidly emerging in apitherapy. Bee products have significant potential to strengthen the immune system and improve human health. Thus, this review is targeted toward recapitulating the chemo-preventive potential of melittin (MEL), which constitutes a substantial portion of honeybee venom. Honeybee venom (apitoxin) is produced in the venom gland of the honeybee abdomen, and adult bees utilize it as a primary colony defense mechanism. Apitoxin comprises numerous biologically active compounds, including peptides, enzymes, amines, amino acids, phospholipids, minerals, carbohydrates, and volatile components. We are mainly focused on exploring the potential of melittin (a peptide component) of bee venom that has shown promising potential in the treatment of several human cancers, including breast, stomach, lung, prostate, ovary, kidney, colon, gastric, esophageal, cervical cancers, melanoma, osteosarcoma, and hepatocellular carcinoma. This review has summarized all potential studies related to the anticancerous efficacy of melittin (apitoxin), its formulations, conjugates, and nano-formulations against several human carcinomas, which would further pave the way for future researchers in developing potent drugs for cancer management. [ABSTRACT FROM AUTHOR]

Honey bees are small insects with many useful products. Venom is one of these products with therapeutic benefits and high commercial value. Many factors affect the bee venom and its main components, namely Melittin, Apamine and Phospholipase A2. This study investigates the effect of seasonal and geographical location on the venom composition in 8 cities, during three seasons of spring, summer and autumn, in a completely randomized design with 4 replications(hives). Analysis of venom samples from the target areas in different seasons by HPLC showed that the mentioned components, other than apamine (correlation=0.339), had no correlation with temperature. The mean amount of Protein, Melittin and Phospholipase A2 of the treatments did not differ significantly during the 3 seasons (p>0.05). The mean percentage of melittin (63.15 %) and phospholipase A2 (8.19 %) in spring; apamine (4.03 %) and protein (48.89 μg/100mg) were maximum in summer. The highest amount of Protein and Apamin in the venom was related to karaj (50.28% and 4.33% respectively), the highest amount of Melittin in khoramabad city and the highest Phospholipase A2 was in birjand city. The effects of season have on venom components through temperature can be in addition to the direct effect on honey bees and venom production glands, through the effect on vegetation of area or the presence of seasonal changes in the venom components in addition to the effect of geographical location, a reflection of the difference in vegetation. [ABSTRACT FROM AUTHOR]

Antibiotic resistance has dramatically spread in recent decades and has become a serious problem in modern medicine. Honeybee venom or bee venom (BV) has anticancer, anti-inflammation, and antimicrobial properties. This study aimed to evaluate the influence of BV on certain pathogenic bacteria. The effects of different concentrations of BV against Klebsiella pneumonia, Pseudomonas aeruginosa, and Staphylococcus aureus were evaluated by disk diffusion method (inhibition zone, IZ), minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Additionally, the antibacterial activity of BV was compared with that of ampicillin, penicillin, and tetracycline. The results indicated that different concentrations of BV had an inhibitory effect on the pathogen species. The MIC and MBC of BV were 18 and 24 μg/mL for K. pneumonia, respectively, 38 and 42 μg/mL for P. aeruginosa, respectively, and 21 and 31 μg/mL for S. aureus, respectively. Among the bacterial species tested, BV was the most effective against K. pneumonia. Meanwhile, P. aeruginosa was the most resistant against BV. The comparison revealed that 100 μg/mL BV had a greater effect on IZ against P. aeruginosa and K. pneumonia compared with evaluated common antibiotics (p < 0.05). This study showed that BV demonstrated antibacterial effects. Precise toxicology examinations are required for the development of new antibiotics. [ABSTRACT FROM AUTHOR]

Background: The composition of venom extracts, cross‐reactive carbohydrate determinants (CCD) and the component‐resolved diagnostics (CRD) are important fields of investigation. IgE‐reactivity to CCD complicates the interpretation of IgE to Hymenoptera venoms, especially in patients with multiple‐positivity. We analyzed the clinical importance of CRD and CCD‐inhibition for selection of allergens for venom immunotherapy (VIT). Methods: In 71 patients, we measured specific IgE (sIgE) to honeybee venom (HBV), wasp venom (WV), hornet venom (HV), CCD, and recombinant allergens: phospholipase A2 (rApi m 1), hyaluronidase (rApi m 2), icarapin (rApi m 10), antigen 5 (rVes v 5), and phospholipase A1 (Immunoblot). In 29/71 HBV/WV/HV/CCD‐positive patients CCD‐inhibition was performed. According to CRD and CCD‐inhibition, we identified true sensitization and defined groups of multiple‐positive patients who needed CCD‐inhibition before starting VIT. Results: sIgE‐rApi m 1, sIgE‐rApi m 2, and sIgE‐rApi m 10 were detected in 65.7%, 68.4%, and 58%, respectively. In HBV allergic patients, CRD sensitivity was 86.8%. In WV allergic patients, sensitivity of sIgE‐rVes v 5 was 94%. True multiple‐sensitization was found in 44.8% of HBV/WV/HV/CCD‐positive patients after CCD‐inhibition. Patients with multiple venom‐ and CCD‐positivity had more frequent severe allergic reactions (p < 0.001). CCD‐inhibition was helpful in HBV/WV/HV/CCD‐positive patients who were negative to all tested recombinant honeybee allergens. Persistence of HBV‐positivity after CCD‐inhibition requires CRD to other honeybee recombinant allergens. Conclusion: CRD, using a profile of five most important recombinant allergens and CCD, has a high sensitivity for the diagnosis of venom allergy, especially in patients positive to several venom extracts. CRD and CCD‐inhibition are helpful to reveal the clinically relevant, true sensitization and improve the selection of venoms for long‐lasting VIT. [ABSTRACT FROM AUTHOR]

Melittin, the most potent pharmacological ingredient of honey bee venom, induces haemolysis, lymphocyte lysis, long-term pain, localised inflammation, and hyperalgesia. In this study, efforts were made to subdue the melittin’s ill effects using a chaperone peptide called ‘mini-αA-crystallin’ (MAC) derived from eye lens αA-crystallin. Haemolytic test on human red blood cells, percentage viability, and DNA diffusion assay on Human peripheral blood lymphocytes (HPBLs) were performed with melittin in the presence or absence of MAC. Propidium iodide and Annexin V-FITC dual staining were performed to analyse quantitative levels of necrotic and apoptotic induction by melittin in the presence or absence of MAC on HPBLs using a flow cytometer. A computational study to find out the interactions between MAC and melittin was undertaken by modelling the structure of MAC using a PEP-FOLD server. The result showed that MAC inhibited melittin-induced lysis in nucleated (lymphocytes) and enucleated (RBC) cells. Flow cytometric analysis revealed a substantial increase in the necrotic and late apoptotic cells after treating HPBLs with melittin (4 µg/ml) for 24 h. Treatment with MAC at a 2:1 molar ratio prevented HPBLs from developing melittin-induced necrosis and late apoptosis. In the docking study, hydrogen, van der Waals, π-π stacking, and salt bridges were observed between the MAC and melittin complex, confirming a strong interaction between them. The MAC-melittin complex was stable during molecular dynamics simulation. These findings may be beneficial in developing a medication for treating severe cases of honeybee stings. [ABSTRACT FROM AUTHOR]

Honeybee venom (HBV) is a product of natural origin with an extremely complex composition. It exhibits several beneficial pharmacological properties, for example anti-inflammatory, antimicrobial, antidiabetic, anti-arthritic, and anti-tumor activities. On the other hand, HBV contains highly allergenic components, including melittin, which makes up more than 50% of its dry weight. Despite many studies conducted, HBV's components and mechanism of action are not yet fully known and understood. Therefore, to enhance the safety of its therapeutical applications, it is essential to establish HBV's composition and biological properties. Many analytical techniques and methods have been reported as potentially useful for the determination of new HBV constituents. Studies focus mainly on the analyses of the HBV proteome and metabolome. Also, the properties of its components when applied to the skin, as well as its allergenicity, are being studied. Currently, the most commonly used methods for HBV analysis include high-performance liquid chromatography and mass spectrometry. Choosing the correct and most applicable method for HBV analysis is of great importance in order to obtain in-depth results crucial for understanding of its mechanism of biological action. The diversity of chemical compounds in HBV also entails an appropriate sample preparation at the pre-analytical stage. In order to summarise the latest knowledge on techniques for the analysis of complex natural products, on the example of HBV, this review paper discusses the modern analytical tools of HBV characterization, focusing on metabolomic and proteomic studies, topical applications, investigation of venom allergies, and the prospects for future research. [ABSTRACT FROM AUTHOR]

Abstract Background Honeybees are one of the most important pollinators in the world, and their products are nowadays included in most anticancer, antiallergic, antimicrobial drugs and are included in cosmetic treatments. In the present study, honeybee venom and Egyptian ethanolic propolis extract (EP) were focused to test their effect on health and some genes for honeybee workers (defensin2, abaecin, hymenoptaecin, vitellogenin, and juvenile hormone esterase). Results Honeybee venom and Egyptian propolis extract (EP) were used as supplements in the nutrition with different concentrations in Varroa mites-infected colonies to measure the colonies' activities after treatment. The immune-related genes and antimicrobial peptides (AMPs) were evaluated by using qRT-PCR. Treated colonies with HBV and EP showed up-regulation of immune and immune-related genes' expressions and increased the life span, activities and their density of bee workers. The data illustrated that the highest gene expression fold of juvenile hormone esterase was detected in the treated colonies with Egyptian ethanolic propolis extract (EP), while the highest vitellogenin expression fold in treated colonies was with honeybee venom. The up-regulation of antimicrobial peptides occurred in colonies with both treatments. Conclusion The findings suggest that honeybee venom and Egyptian ethanolic propolis extract (EP) could be used as potential supplements, even at the lowest concentration to develop the immunity of worker bees to increase their efficiency and prevent loss of honeybee colonies due to several diseases closely associated with Varroa mites' infections that cause sudden death.

Honeybee venom (HBV) is an important product of beehives, and its benefits for health have been rediscovered by modern medicine. Since HBV has the potential to treat some diseases, its quality and production conditions require a detailed investigation. The objective of this study is to understand how season, harvesting time (day or night), harvesting site of beehives (inside or entrance) and geographic location affects quality through the analysis of apamin, melittin and phospholipase A2 (PLA2) content. Each set of six colonies were used to understand the differences in these components when HBV is harvested in the daytime either from the entrance or inside of the hives and nighttime from the inside of the hives. The experiment also investigated seasonal differences as the samples were harvested each month from May to August 2019 in our apiary. Furthermore, the effect of geographic location on quality was examined through the comparison of the data obtained from twenty-seven samples collected by beekeepers using the same device, located in the Turkish cities of Manisa, Muğla, Balıkesir, Düzce and Mersin. The results demonstrated that statistically significant differences in the amounts of analyzed components were not dependent on harvesting time, collection site on the beehives or season. On the other hand, region samples significantly differed in the amounts of all three components, ranging from 1.28% to 3.81% for apamin, 19.51–64.03% for melittin and 7.22%–28.18% for PLA2. However, beekeepers’ improper practices during harvesting and storing might be the most critical parameters that determine the quality of HBV.

Abstract Background The composition of venom extracts, cross‐reactive carbohydrate determinants (CCD) and the component‐resolved diagnostics (CRD) are important fields of investigation. IgE‐reactivity to CCD complicates the interpretation of IgE to Hymenoptera venoms, especially in patients with multiple‐positivity. We analyzed the clinical importance of CRD and CCD‐inhibition for selection of allergens for venom immunotherapy (VIT). Methods In 71 patients, we measured specific IgE (sIgE) to honeybee venom (HBV), wasp venom (WV), hornet venom (HV), CCD, and recombinant allergens: phospholipase A2 (rApi m 1), hyaluronidase (rApi m 2), icarapin (rApi m 10), antigen 5 (rVes v 5), and phospholipase A1 (Immunoblot). In 29/71 HBV/WV/HV/CCD‐positive patients CCD‐inhibition was performed. According to CRD and CCD‐inhibition, we identified true sensitization and defined groups of multiple‐positive patients who needed CCD‐inhibition before starting VIT. Results sIgE‐rApi m 1, sIgE‐rApi m 2, and sIgE‐rApi m 10 were detected in 65.7%, 68.4%, and 58%, respectively. In HBV allergic patients, CRD sensitivity was 86.8%. In WV allergic patients, sensitivity of sIgE‐rVes v 5 was 94%. True multiple‐sensitization was found in 44.8% of HBV/WV/HV/CCD‐positive patients after CCD‐inhibition. Patients with multiple venom‐ and CCD‐positivity had more frequent severe allergic reactions (p

Honeybee venom is one of the natural substances produced by bees ( Apis mellifera ). Their venom gland produces venom which plays a defensive role. In this study a concentration of macro and trace elements (Ag, Al, As, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Li, Mg, Mn, Mo, Na, Ni, Sb, Se, Sr, Pb and Zn) in foragers' and honeybees' venom was analysed by axial inductively coupled plasma optical emission spectrometry (ICP OES) with good validation parameters to differentiate the element accumulation ability in honeybee venom. Cumulative ability for some elements (As, Al, Ba, Cr, Li, Mo, Pb, and Zn) in bee venom was clearly demonstrated. Oppositely, levels of macro elements (Ca, K, Mg and Na) in venom were several times lower compared to the levels detected in foragers. Moreover, PCA analysis of bee samples showed that Cr was associated with locality Košice, and Cd with locality Krompachy; both have rich industrial history. Since some of analysed elements are potentially toxic for humans, a risk assessment for bee-stung scenario was also calculated. A new way of exposure to potentially toxic elements via honeybee stung was showed in this study. Non-carcinogenic risk assessment for humans to selected toxic elements (As, Cd, Cr, Ni, and Pb) demonstrated acceptable risk and moreover the same we may conclude for potential carcinogenic risk for beekeepers exposed to As, Cd, Ni, and Pb via venom over their whole life., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors. Published by Elsevier Ltd.)

Apitherapy (using bee products) has gained broad recognition in cancer therapeutics globally. Honeybee venom has a broad range of biological potential, and its utilization is rapidly emerging in apitherapy. Bee products have significant potential to strengthen the immune system and improve human health. Thus, this review is targeted toward recapitulating the chemo-preventive potential of melittin (MEL), which constitutes a substantial portion of honeybee venom. Honeybee venom (apitoxin) is produced in the venom gland of the honeybee abdomen, and adult bees utilize it as a primary colony defense mechanism. Apitoxin comprises numerous biologically active compounds, including peptides, enzymes, amines, amino acids, phospholipids, minerals, carbohydrates, and volatile components. We are mainly focused on exploring the potential of melittin (a peptide component) of bee venom that has shown promising potential in the treatment of several human cancers, including breast, stomach, lung, prostate, ovary, kidney, colon, gastric, esophageal, cervical cancers, melanoma, osteosarcoma, and hepatocellular carcinoma. This review has summarized all potential studies related to the anticancerous efficacy of melittin (apitoxin), its formulations, conjugates, and nano-formulations against several human carcinomas, which would further pave the way for future researchers in developing potent drugs for cancer management.

Background: Honeybees are one of the most important pollinators in the world, and their products are nowadays included in most anticancer, antiallergic, antimicrobial drugs and are included in cosmetic treatments. In the present study, honeybee venom and Egyptian ethanolic propolis extract (EP) were focused to test their effect on health and some genes for honeybee workers (defensin2, abaecin, hymenoptaecin, vitellogenin, and juvenile hormone esterase). Results: Honeybee venom and Egyptian propolis extract (EP) were used as supplements in the nutrition with different concentrations in Varroa mites-infected colonies to measure the colonies' activities after treatment. The immune-related genes and antimicrobial peptides (AMPs) were evaluated by using qRT-PCR. Treated colonies with HBV and EP showed up-regulation of immune and immune-related genes' expressions and increased the life span, activities and their density of bee workers. The data illustrated that the highest gene expression fold of juvenile hormone esterase was detected in the treated colonies with Egyptian ethanolic propolis extract (EP), while the highest vitellogenin expression fold in treated colonies was with honeybee venom. The up-regulation of antimicrobial peptides occurred in colonies with both treatments. Conclusion: The findings suggest that honeybee venom and Egyptian ethanolic propolis extract (EP) could be used as potential supplements, even at the lowest concentration to develop the immunity of worker bees to increase their efficiency and prevent loss of honeybee colonies due to several diseases closely associated with Varroa mites' infections that cause sudden death. [ABSTRACT FROM AUTHOR]

5-fluorouracil (5-FU) and doxorubicin (DOX) are potent anti-tumour agents commonly used for colon and breast cancer therapy, respectively. However, their clinical application is limited by their side effects and the development of drug resistance. Honeybee venom is a complex mixture of substances that has been reported to be effective against different cancer cells. Its active compound is melittin, a positively charged amphipathic peptide that interacts with the phospholipids of the cell membrane, forming pores that enable the internalization of small molecules with cytotoxic activities,. and consequently, causing cell death. Some central nervous system (CNS) drugs have recently demonstrated great anti-cancer potential, both in vitro, in vivo and in clinical trials, being promising candidates for drug repurposing in oncology. The present work evaluated the anti-cancer efficacy of honeybee venom in combination with chemotherapeutic or CNS drugs in HT-29 colon and MCF-7 breast cancer cell lines. The chemical characterization of a Portuguese sample of honeybee venom was done by LC-DAD-ESI/MSn analysis. For single treatments, cells were incubated with increasing concentrations of bee venom. For combination treatments, increasing concentrations of bee venom were first combined with the half-maximal inhibitory concentration (IC50) of 5-FU and DOX, in HT-29 and MCF-7 cells, respectively. Cells were also treated with increasing concentrations of bee venom in combination with the IC50 value of four CNS drugs (fluphenazine, fluoxetine, sertraline and thioridazine). Cytotoxicity was evaluated by MTT and SRB assays. The combination index (CI) value was calculated using CompuSyn software, based on the Chou–Talalay method. Synergy scores of different reference models (HSA, Loewe, ZIP and Bliss) were also calculated using SynergyFinder. The results demonstrate that honeybee venom is active against HT-29 colon and MCF-7 breast cancer cells, having better anti-tumour activity in MCF-7 cells. It was found that bee venom combined with 5-FU and fluphenazine in HT-29 cells resulted in less cytotoxic effects compared to the co-treatment of fluoxetine, sertraline and thioridazine plus bee venom, which resulted in less than 15% of viable cells for the whole range of concentrations. The combination of MCF-7 cells with repurposed drugs plus honeybee venom resulted in better anti-cancer efficacies than with DOX, notably for lower concentrations. A combination of fluoxetine and thioridazine plus honeybee venom resulted in less than 40% of viable cells for all ranges of concentrations. These results support that the combination of honeybee venom with repurposed drugs and chemotherapeutic agents can help improve their anti-cancer activity, especially for lower concentrations, in both cell lines. Overall, the present study corroborates the enormous bioactive potential of honeybee venom for colon and breast cancer treatments, both alone and in combination with chemotherapy or repurposed drugs. [ABSTRACT FROM AUTHOR]

Background: In contrast to their clearly defined roles in allergic diseases, the physiologic functions of Immunoglobulin E antibodies (IgEs) and mast cells (MCs) remain enigmatic. Recent research supports the toxin hypothesis, showing that MCs and IgE‐related type 2 immune responses can enhance host defense against certain noxious substances, including honeybee venom (BV). However, the mechanisms by which MCs can interfere with BV toxicity are unknown. In this study, we assessed the role of IgE and certain MC products in MC‐mediated BV detoxification. Methods: We applied in vitro and in vivo fluorescence microscopyimaging, and flow cytometry, fibroblast‐based toxicity assays and mass spectrometry to investigate IgE‐mediated detoxification of BV cytotoxicity by mouse and human MCs in vitro. Pharmacologic strategies to interfere with MC‐derived heparin and proteases helped to define the importance of specific detoxification mechanisms. Results: Venom‐specific IgE increased the degranulation and cytokine responses of MCs to BV in vitro. Passive serum sensitization enhanced MC degranulation in vivo. IgE‐activated mouse or human MCs exhibited enhanced potential for detoxifying BV by both proteolytic degradation and heparin‐related interference with toxicity. Mediators released by IgE‐activated human MCs efficiently degraded multiple BV toxins. Conclusions: Our results both reveal that IgE sensitization enhances the MC's ability to detoxify BV and also assign efficient toxin‐neutralizing activity to MC‐derived heparin and proteases. Our study thus highlights the potential importance of IgE, MCs, and particular MC products in defense against BV. [ABSTRACT FROM AUTHOR]

Hymenopteran insect stings are a risk factor that cannot be ignored for the people allergic to hymenopteran venoms.In China,the current diagnostic tools cannot provide accurate information to identify sensitized insects,thus affecting clinical diagnosis and treatment.Honeybee is a common hymenopteran insect.Due to its wide distribution,large number,and complex venom composition,researchers have carried out recombination schemes for the main allergens of honeybee venom,laying a theoretical foundation for the detection of allergens.The development of diagnostic technologies for allergen components can accurately detect bee venom allergens,providing a new set of clinical diagnosis and treatment schemes for the population allergic to bee venom.

Objective: Salmonella Typhimurium is a significant zoonotic concern for human food poisoning and a substantial economic burden in the swine industry. We previously reported that nasally delivered chitosan-coated poly(lactide-co-glycolide) (PLGA) encapsulating honeybee venom (CP-HBV) could enhance CD4+ T helper 1 (Th1)-related immune responses in healthy pigs. Building upon these findings, the current study aimed to investigate the protective immune enhancement by nasally delivered CP-HBV in pigs challenged with S Typhimurium., Animals: 36 healthy, 4-week-old, female, Landrace X Yorkshire X Duroc pigs., Methods: 36 pigs were allocated into 3 groups: CP-HBV (n = 16), control (n = 16), and healthy baseline control (n = 4). CP-HBV and control groups were challenged with S Typhimurium 7 days post-treatment. Pigs from the healthy control group were sacrificed at 0 days postinfection (DPI), and 4 pigs from each of the control and CP-HBV groups were sacrificed at 1, 2, 4, and 7 DPI. Salmonella shedding, immune cell frequencies, cytokines, and transcriptional factor expression levels were measured., Results: The CP-HBV group exhibited lower bacterial shedding and an enhanced Th1-related immune response characterized by an upregulation of CD4+ T cells and CD4+ Interferon-γ+ T cells, accompanied by increased expression of Th1-related cytokines and reduced expression of regulatory T cells and immunosuppressive cytokines compared to the control group., Clinical Relevance: CP-HBV is a promising strategy for controlling Salmonella infections in pigs and improving public health.

Background: Breast cancer is the most common and life-threatening cancer in females characterized by the abnormal proliferation of tumor cells in lobules and ducts. For years, many anti-breast cancer drugs have been tested with some of them showing severe health problems and drug resistance. Recently, different biological and pharmacological actions of bee venom have been indicated to play antibacterial, anti-viral and anti-inflammatory role against different cancers specially breast cancer. Methods: This review study is based on PubMed, Google Scholar and PubMed search. Search terms used were Melittin, Breast cancer and Honey Bee Venom. Results: Many studies have shown that a positively charged C-terminal sequence of mellitin facilitates plasma membrane contact and antitumor action. Precise targeting and selective activity of melittin has been found in recent studies as it suppresses the activation of growth factor receptors in HER2-enriched and triplenegative breast cancer that are generally difficult to treat. Significantly, it leaves healthy cells intact. The most striking feature of melittin is the pore formation property. Monomers of melittin bind to the plasma membrane of cancer cells in a collective manner and start forming pores ultimately bringing cell lysis. Conclusion: Since melittin has a very selective action against the HER-2 related tumors, a combinational therapy of melittin and HER-2 targeted agents could be a very potent strategy in breast cancer. This review reflects the importance of honey bee venom and melittin as a potential therapy for aggressive breast cancer. [ABSTRACT FROM AUTHOR]

Background: Breast cancer is the most common and life-threatening cancer in females characterized by the abnormal proliferation of tumor cells in lobules and ducts. For years, many anti-breast cancer drugs have been tested with some of them showing severe health problems and drug resistance. Recently, different biological and pharmacological actions of bee venom have been indicated to play anti-bacterial, anti-viral and anti-inflammatory role against different cancers especially breast cancer. Methods: This review study is based on PubMed, Google Scholar and PubMed search. Search terms used were Melittin, Breast cancer and Honey Bee Venom. Results: Many studies have shown that a positively charged C-terminal sequence of mellitin facilitates plasma membrane contact and antitumor action. Precise targeting and selective activity of melittin has been found in recent studies as it suppresses the activation of growth factor receptors in HER2-enriched and triple-negative breast cancer that are generally difficult to treat. Significantly, it leaves healthy cells intact. The most striking feature of melittin is the pore formation property. Monomers of melittin bind to the plasma membrane of cancer cells in a collective manner and start forming pores, ultimately bringing cell lysis. Conclusion: Since melittin has a very selective action against the HER-2 related tumors, a combinational therapy of melittin and HER-2 targeted agents could be a very potent strategy in breast cancer. This review reflects the importance of honey bee venom and melittin as a potential therapy for aggressive breast cancer.

The treatment of diseases with honeybee venom (apitoxin) is a branch of apitherapy, which has yet to find substantial scientific support. In this present study, we used apitoxin to investigate its effects on diabetes mellitus (DM) in rats. The use of apitoxin injection on single-dose streptozotocin (STZ)-treated rat model was selected, and the study took 28 days. Fasting blood glucose (FBG) was measured weekly. Biochemical and hematological parameters, oxidative stress markers, and insulin levels were measured from the blood samples provided on the last day of the study, and the DNA damage marker (8-OHdG) was measured from urine samples. We found that apitoxin increased malondialdehyde (MDA) and carbonylated protein (PCO), and decreased glutathione (GSH) levels significantly. Additionally, apitoxin increased nitric oxide (NOx) levels significantly in diabetic rats. The findings suggest that apitoxin increases oxidative stress to some extent. Apitoxin had no significant effect on plasma insulin levels. Apitoxin apparently caused small decreases in FBG levels. Apitoxin appears to cause an increase in aspartate aminotransferase (AST), and an increase in the number of platelets (PLT) and granulocytes (GRAN) in diabetic rats. Consequently, the possible antidiabetic effect of apitoxin on DM could only be supported at the preliminary level and presented with speculation in this state of the study. [ABSTRACT FROM AUTHOR]

Hypercholesterolemia can be defined as the presence of high plasma cholesterol levels, with normal plasma triglycerides, as a consequence of the rise of cholesterol and apolipoprotein B (apoB)-rich lipoproteins, called low-density lipoprotein (LDL) and characterized by very high levels on the blood cholesterol which is a risk factor of cardiovascular health. Medicines which formed from chemical compounds cause serious side effects that cause an imbalance in the body's functions. Therefore, using of natural substances may be an innovative method for the treatment of diseases, as they do not have any side effects on human health. Bee venom can be used as a drug due to its beneficial effects on many disorders, such as neurological, cardiovascular, hematological, musculoskeletal, and dermatological disorders. In the present work, we found that treatment of hypercholesterolemic rats with bee venom (BV) revealed significant increases in Hb, Hct, and RBCs count and showed a significant increase in PaO2, % O2 saturation, O2 carrying capacity. These results suggest that BV can enhance respiratory functions of blood and can be used in respiratory diseases to enhance the respiration and hematological parameters. Also, BV could be used as a prophylactic agent (food protector) to protect us from the harmful effects of cholesterol. [ABSTRACT FROM AUTHOR]

A new method for determination of selected heavy metals (Cd, Pb, Cu, Zn, and Ni) in honey bee venom was developed. Heavy metals are metabolized and incorporated into bee products, including honey and honey bee venom (apitoxin). Their composition reflects contamination of "bee environment", providing information about heavy metal contamination in the neighborhood of human dwellings. Moreover, assessment of bee products contamination is relevant for medicine, as they are a tool for promising therapeutic and chemoprophylactic strategies against COVID-19 (SARS-CoV-2). Owing to the complicated matrix, the developed method consists of wet mineralization with sulfuric acid, nitric acid, under increased temperature, and pressure and subsequent repeated boiling with concentrated nitric acid. Determination of the selected metals was carried out by anodic or cathodic stripping voltammetry on two types of electrodes: pen-type hanging mercury drop electrode (HMDE) and PLA filament with carbon conductive admixture (PLA-C) for 3D printer. Contents of lead and cadmium in all analyzed bee venom samples were on the level of mg kg−1, of nickel and copper about ten times higher, and of zinc on the level of g kg−1. The results achieved using HMDE were recorded with average relative standard deviation (RSD) 5.4% (from 3.2% to 8.6%) and using PLA-C 11.8% (from 6.5% to 18.0%). The results achieved using both electrodes proved to be equivalent with statistical probability higher than 95%. [ABSTRACT FROM AUTHOR]

5-fluorouracil (5-FU) and doxorubicin (DOX) are potent anti-tumour agents commonly used for colon and breast cancer therapy, respectively. However, their clinical application is limited by their side effects and the development of drug resistance. Honeybee venom is a complex mixture of substances that has been reported to be effective against different cancer cells. Its active compound is melittin, a positively charged amphipathic peptide that interacts with the phospholipids of the cell membrane, forming pores that enable the internalization of small molecules with cytotoxic activities,. and consequently, causing cell death. Some central nervous system (CNS) drugs have recently demonstrated great anti-cancer potential, both in vitro, in vivo and in clinical trials, being promising candidates for drug repurposing in oncology. The present work evaluated the anti-cancer efficacy of honeybee venom in combination with chemotherapeutic or CNS drugs in HT-29 colon and MCF-7 breast cancer cell lines. The chemical characterization of a Portuguese sample of honeybee venom was done by LC-DAD-ESI/MSn analysis. For single treatments, cells were incubated with increasing concentrations of bee venom. For combination treatments, increasing concentrations of bee venom were first combined with the half-maximal inhibitory concentration (IC50) of 5-FU and DOX, in HT-29 and MCF-7 cells, respectively. Cells were also treated with increasing concentrations of bee venom in combination with the IC50 value of four CNS drugs (fluphenazine, fluoxetine, sertraline and thioridazine). Cytotoxicity was evaluated by MTT and SRB assays. The combination index (CI) value was calculated using CompuSyn software, based on the Chou–Talalay method. Synergy scores of different reference models (HSA, Loewe, ZIP and Bliss) were also calculated using SynergyFinder. The results demonstrate that honeybee venom is active against HT-29 colon and MCF-7 breast cancer cells, having better anti-tumour activity in MCF-7 cells. It was found that bee venom combined with 5-FU and fluphenazine in HT-29 cells resulted in less cytotoxic effects compared to the co-treatment of fluoxetine, sertraline and thioridazine plus bee venom, which resulted in less than 15% of viable cells for the whole range of concentrations. The combination of MCF-7 cells with repurposed drugs plus honeybee venom resulted in better anti-cancer efficacies than with DOX, notably for lower concentrations. A combination of fluoxetine and thioridazine plus honeybee venom resulted in less than 40% of viable cells for all ranges of concentrations. These results support that the combination of honeybee venom with repurposed drugs and chemotherapeutic agents can help improve their anti-cancer activity, especially for lower concentrations, in both cell lines. Overall, the present study corroborates the enormous bioactive potential of honeybee venom for colon and breast cancer treatments, both alone and in combination with chemotherapy or repurposed drugs.

Propolis is a resinous honeybee product, rich in polyphenolic compounds and with high economic value. Although extensive studies regarding the chemical composition of different propolis extracts have been carried out, the propolis proteome remained unknown. The present study aimed at characterizing the proteome of two geographically-distinct propolis originating from Belgium and Iran. Propolis extracts were analyzed by sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE) followed by Matrix Assisted Laser Desorption Ionization – Time of Flight Mass Spectrometry (MALDI-TOF MS) and Liquid chromatography-electrospray ionization MS (LC-ESI-MS) analyses. Our comprehensive proteomic analysis led to the identification of honeybee venom and royal jelly proteins, and numerous plant-defense proteins in propolis, extending our understanding of the antimicrobial properties of this complex natural substance. The proteome of both propolis was mainly made up of proteins belonging to the poplar tree. Pathogenesis-related (PR) proteins, poplar PR-2 (β-1,3-glucanases) and PR-8 (acidic endochitinases) were found to be the major components of both propolis extracts, while the major bee venom allergen (Api m 1) and royal jelly proteins (MJRP 1, 2, 3 and 5) were identified in the propolis from Belgium. Overall, our proteome analyses revealed that propolis extracts from Belgium and Iran shared a core protein composition originating from the poplar proteome. The majority of identified proteins are involved in the plant defense against pathogens, most belonging to well-known pathogenesis-related families. Proteomics could thus represent a viable approach for origin authentication and allergen identification in poplar-type propolis. • First proteomic analysis on propolis using two geographically distinct samples. • ESI- and MALDI-MS led to the identification of 74 non-redundant protein hits. • The proteome of both propolis samples was mainly composed of poplar proteins. • Two samples shared a core protein composition (poplar PR-1, 2, 3, 4, 5, 8 and 14). • Authentication of propolis and allergenic risk assessments are major applications. [ABSTRACT FROM AUTHOR]

In this review, we outline and reflect on the important differences between allergen-specific immunotherapy for inhalant allergies (i.e., aeroallergens) and venom-specific immunotherapy (VIT), with a special focus on Venomil® Bee and Wasp. Venomil® is provided as a freeze-dried extract and a diluent to prepare a solution for injection for the treatment of patients with IgE-mediated allergies to bee and/or wasp venom and for evaluating the degree of sensitivity in a skin test. While the materials that make up the product have not changed, the suppliers of raw materials have changed over the years. Here, we consolidate relevant historical safety and efficacy studies that used products from shared manufacture supply profiles, i.e., products from Bayer or Hollister–Stier. We also consider the characterization and standardization of venom marker allergens, providing insights into manufacturing controls that have produced stable and consistent quality profiles over many years. Quality differences between products and their impacts on treatment outcomes have been a current topic of discussion and further research. Finally, we review the considerations surrounding the choice of depot adjuvant most suitable to augmenting VIT.

Stings of hymenoptera can induce IgE-mediated hypersensitivity reactions in venom-allergic patients, ranging from local up to severe systemic reactions and even fatal anaphylaxis. Allergic patients' quality of life can be mainly improved by altering their immune response to tolerate the venoms by injecting increasing venom doses over years. This venom-specific immunotherapy is highly effective and well tolerated. However, component-resolved information about the venoms has increased in the last years. This knowledge is not only able to improve diagnostics as basis for an accurate therapy, but was additionally used to create tools which enable the analysis of therapeutic venom extracts on a molecular level. Therefore, during the last decade the detailed knowledge of the allergen composition of hymenoptera venoms has substantially improved diagnosis and therapy of venom allergy. This review focuses on state of the art diagnostic and therapeutic options as well as on novel directions trying to improve therapy.

Massive, Africanized honeybee attacks have increased in Brazil over the years. Humans and animals present local and systemic effects after envenomation, and there is no specific treatment for this potentially lethal event. This study evaluated the ability of a new Apilic antivenom, which is composed of F(ab’)2 fraction of specific immunoglobulins in heterologous and hyperimmune equine serum, to neutralize A. mellifera venom and melittin, in vitro and in vivo, in mice. Animal experiments were performed in according with local ethics committee license (UFRJ protocol no. DFBCICB072-04/16). Venom dose-dependent lethality was diminished with 0.25–0.5 μL of intravenous Apilic antivenom/μg honeybee venom. In vivo injection of 0.1–1 μg/g bee venom induced myotoxicity, hemoconcentration, paw edema, and increase of vascular permeability which were antagonized by Apilic antivenom. Cytotoxicity, assessed in renal LLC-PK1 cells and challenged with 10 μg/mL honeybee venom or melittin, was neutralized by preincubation with Apilic antivenom, as well the hemolytic activity. Apilic antivenom inhibited phospholipase and hyaluronidase enzymatic activities. In flow cytometry experiments, Apilic antivenom neutralized reduction of cell viability due to necrosis by honeybee venom or melittin. These results showed that this antivenom is effective inhibitor of honeybee venom actions. Thus, this next generation of Apilic antivenom emerges as a new promising immunobiological product for the treatment of massive, Africanized honeybee attacks.

The aim of this study was to explore the serum peptide profiles from honeybee stung and non-stung individuals. Two groups of serum samples obtained from 27 beekeepers were included in our study. The first group of samples was collected within 3 h after a bee sting (stung beekeepers), and the samples were collected from the same person a second time after at least six weeks after the last bee sting (non-stung beekeepers). Peptide profile spectra were determined using MALDI-TOF mass spectrometry combined with Omix, ZipTips and magnetic beads based on weak-cation exchange (MB-WCX) enrichment strategies in the mass range of 1–10 kDa. The samples were classified, and discriminative models were established by using the quick classifier, genetic algorithm and supervised neural network algorithms. All of the statistical algorithms used in this study allow distinguishing analyzed groups with high statistical significance, which confirms the influence of honeybee sting on the serum peptidome profile. The results of this study may broaden the understanding of the human organism’s response to honeybee venom. Due to the fact that our pilot study was carried out on relatively small datasets, it is necessary to conduct further proteomic research of the response to honeybee sting on a larger group of samples.

Honeybee ( Apis mellifera ) venom (HBV) represents an ideal model to study the role of particular venom components in allergic reactions in sensitized individuals as well as in the eusociality of Hymenoptera species. The aim of this study was to further characterize the HBV components C1q-like protein (C1q) and PDGF/VEGF-like factor 1 (PVF1). C1q and PVF1 were produced as recombinant proteins in insect cells. Their allergenic properties were examined by determining the level of specific IgE antibodies in the sera of HBV-allergic patients (n = 26) as well as by their capacity to activate patients' basophils (n = 11). Moreover, the transcript heterogeneity of PVF1 was analyzed. It could be demonstrated that at least three PVF1 variants are present in the venom gland, which all result from alternative splicing of one transcript. Additionally, recombinant C1q and PVF1 from Spodoptera frugiperda insect cells exhibited specific IgE reactivity with approximately 38.5% of sera of HBV-allergic patients. Interestingly, both proteins were unable to activate basophils of the patients, questioning their role in the context of clinically relevant sensitization. Recombinant C1q and PVF1 can build the basis for a deeper understanding of the molecular mechanisms of Hymenoptera venoms. Moreover, the conflicting results between IgE sensitization and lack of basophil activation, might in the future contribute to the identification of factors that determine the allergenic potential of proteins. [ABSTRACT FROM AUTHOR]

Bee stings represent a public health subject, but the mechanisms involved in bee venom toxicity are not yet fully understood. To evaluate the reactions of adrenocortical cells, through which organisms respond to stress, two honeybee venom components: melittin (Mlt) and phospholipase A2 (PLA2) were tested as potential chemical stressors. Modifications were investigated with transmission electron microscopy and microanalysis. A single dose of Mlt (31 mg/kg) or PLA2 (9.3 mg/kg) was injected in rats of groups ML and PL; daily doses of Mlt (350 μg/kg) or PLA2 (105 μg/kg) were injected 30 days in rats of groups M30 and P30. Adrenocortical cells in ML group showed ultrastructural degenerative alterations of nuclei, endoplasmic reticulum, and mitochondria that exhibited lipid inclusions and mitochondrial cristae (MC) re-organized into mono- or multimembrane large vesicles, and whorls of membranes. Many MC were degenerated. In the M30 group, similar ultrastructural changes, but of lower amplitude were noted; lipid cytosolic droplets were heterogenous. MC diameters in Mlt groups (melittin treated groups) were significantly higher than in control (C) group. In PL group, mitochondria contained large lipid inclusions, vesicular MC of different sizes and multiple membranes, and debris, or whorl structures. In P30 group MC were tubular with increased diameters. In both PLA2 groups (PLA2 treated groups) MC were significantly larger than in C group. We concluded that Mlt and PLA2 were powerful stressors, toxic at the tested doses, cellular reactions concerning in all groups mainly mitochondria, but also other cellular compartments. Apart from degenerative regression of MC, the rearrangement of tubular MC occurred into one or multiple large multimembrane vesicular MC. Reactions to the high doses were more pronounced, with the highest amplitude in ML group, and the lowest in P30 group. [Display omitted] • Honeybee venom melittin and phospholipase A2 injected in rats in single high doses and 30 days in daily low doses. • Melittin high dose: polymorphous mitochondria, lipid inclusions, degenerated or extralarge multimembranous cristae. • Melittin 30: similar changes; cristae with significantly lower diameter. • Phospholipase A2 high dose: matrix lipid inclusions, degenerated/vesicular cristae with membranous whorl structures. • Phospholipase A2 30: less severe alterations, larger tubular cristae with tendency to vesiculate. [ABSTRACT FROM AUTHOR]

A reverse-phase high-performance liquid chromatographic (RP -HPLC ) method was developed and validated for the analysis of honeybee venom samples and drug products containing honeybee venom. The validation parameters were linearity, sensitivity, precision, and recovery. Melittin is the main component of honeybee venom was extracted with pure water, and then evaluated by RP -HPLC with a photodiode array (PDA ) detector. Separation of the samples was achieved on a Europa Protein C18 column with linear gradient elution of acetonitrile and 0.4% phosphoric acid at 25°C. There was a flow rate of 1 mL/min. Detection was set at 220 nm. Limits of detection (LOD ) and quantification (LO Q) for melittin were 1.1 and 3.2 μg/mL, respectively. The amount of melittin in honeybee venom samples ranged from 21.9 to 66.4 %.

Our previous study demonstrated that the secapin peptide from the venom of the Asiatic honeybee ( Apis cerana , AcSecapin-1) exhibits anti-fibrinolytic, anti-elastolytic, and anti-microbial activities. In the present study, we investigated the anti-microbial activity and cytotoxicity of a synthetic AcSecapin-1 peptide. Seven synthetic AcSecapin-1 peptides (AcSecapin-S1 to AcSecapin-S7) were synthesized based on the peptide sequence of AcSecapin-1. AcSecapin-S1, which consists of the 25-amino acid sequence identical to that of the mature AcSecapin-1 peptide, exhibited the highest anti-microbial activity against bacteria and fungi. This was followed by AcSecapin-S6, which was missing 10 N-terminal amino acids and 6 C-terminal amino acids from AcSecapin-S1. Furthermore, AcSecapin-S1 was not cytotoxic and did not activate macrophages. Taken together, our data demonstrated that a synthetic AcSecapin-1 peptide could exhibit anti-microbial activity without producing a cytotoxic or inflammatory response, suggesting that this synthetic AcSecapin-1 peptide can be used as an anti-microbial agent for biomedical applications. [ABSTRACT FROM AUTHOR]

Background: Diabetes is an important disease. This disease is a metabolic disorder characterized by hyperglycemia resulting from perturbation in insulin secretion, insulin action or both. Honey bee venom contains a wide range of polypeptide agents. The principle components of bee venom are mellitin and phospholipase A2. These components increase insulin secretion from the β-cells of pancreas. This study was conducted to show the hypoglycemic effect of honey bee venom on alloxan induced diabetic male rats. Methods: Eighteen adult male rats weighting 200±20 g were placed into 3 randomly groups: control, alloxan mono- hydrate-induced diabetic rat and treated group that received honey bee venom daily before their nutrition for four months. Forty eight hours after the last injection, blood was collected from their heart, serum was dissented and blood glucose, insulin, triglyceride and total cholesterol were determined. Results: Glucose serum, triglyceride and total cholesterol level in treated group in comparison with diabetic group was significantly decreased (P< 0.01). On the other hand, using bee venom causes increase in insulin serum in com- parison with diabetic group (P< 0.05). Conclusion: Honeybee venom (apitoxin) can be used as therapeutic option to lower blood glucose and lipids in dia- betic rats.

Nasal delivery is a convenient and acceptable route for drug administration, and has been shown to elicit a much more potent local and systemic response compared with other drug delivery routes. We previously demonstrated that rectal administration of poly(lactide- co -glycolide)-encapsulated honeybee venom (P-HBV) could enhance systemic Th 1-specific immune responses. We therefore synthesized chitosan-coated P-HBV (CP-HBV) and then evaluated the immune-boosting efficacy of nasally administered CP-HBV on systemic and local intestinal immunity compared with non-chitosan-coated P-HBV. The nasally delivered CP-HBV effectively enhanced Th 1-specific responses, eliciting a significant increase in the CD3 + CD4 + CD8 − Th cell population, lymphocyte proliferation capacity, and expression of Th 1 cytokines (IFN-γ, IL-12, and IL-2) in peripheral blood mononuclear cells. Furthermore, these immune-boosting effects persisted up to 21 days post CP-HBV administration. Nasal administration of CP-HBV also led to an increase of not only the CD4 + Th 1 and IFN-γ secreting CD4 + Th 1 cell population but also Th 1-specific cytokines and transcription factors, including IL-12, IFN-γ, STAT4, and T-bet, in isolated mononuclear cells from the spleen and ileum. [ABSTRACT FROM AUTHOR]