ABSTRACT
Botulism is one of the most serious food intoxications, manifesting as prolonged paralytic conditions. This disease is usually the result of the consumption of poor quality canned or smoked foods, so the inhabitants of many countries of the world are exposed to the risk of this kind of poisoning every year. In view of the severity of poisonings caused by botulinum neurotoxins, monoclonal antibodies (mAbs) show great promise because of their targeting action, lack of allergic reactions and serum sickness. The use of a cocktail of mAbs increases the "functional specificity" of their mixture, allowing them to bind to the active domains of different toxin chains and block their action. In this work, we obtained 14 murine mAbs to the catalytic and receptor-binding domain of botulinum toxin type A. The Sp2/0-Ag14 murine myeloma cell line and splenocytes from immunized mice of the BALB/c line were used as fusion partners. We have shown that the selected cocktail of three antibodies neutralizes native toxin more effectively than antibodies separately-complete neutralization is achieved at a toxin dose of 3LD50 and partial neutralization at 5LD50. We presume that this cocktail may be promising as a prototype for the creation of a therapeutic drug capable of neutralizing the toxin in the blood of patients.
Subject(s)
Antibodies, Monoclonal , Botulinum Toxins, Type A , Mice, Inbred BALB C , Animals , Antibodies, Monoclonal/therapeutic use , Antibodies, Monoclonal/immunology , Botulinum Toxins, Type A/immunology , Botulinum Toxins, Type A/toxicity , Botulism/drug therapy , Botulism/immunology , Mice , Cell Line, Tumor , Female , Antibodies, Neutralizing/immunologyABSTRACT
Live anthrax vaccine containing spores from attenuated strains STI-1 of Bacillus anthracis is used in Russia and former CIS (Commonwealth of Independent States) to prevent anthrax. In this paper we studied the duration of circulation of antibodies specific to spore antigens, the protective antigen (PA), the lethal factor (LF) and their domains (D) in donors' blood at different times after their immunization with live anthrax vaccine. The relationship between the toxin neutralization activity level and the level of antibodies to PA, LF and their domains was tested. The effect of age, gender and number of vaccinations on the level of adaptive post-vaccination immune response has been studied. It was shown that antibodies against PA-D1 circulate in the blood of donors for 1 year or more after immunization with live anthrax vaccine. Antibodies against all domains of LF and PA-D4 were detected in 11 months after vaccination. Antibodies against the spores were detected in 8 months after vaccination. A moderate positive correlation was found between the titers of antibodies to PA, LF, or their domains, and the TNA of the samples of blood serum from the donors.
Subject(s)
Adaptive Immunity , Anthrax Vaccines/immunology , Anthrax/immunology , Anthrax/prevention & control , Anthrax Vaccines/administration & dosage , Antibodies, Bacterial/blood , Antibodies, Bacterial/immunology , Antigens, Bacterial/immunology , Bacterial Toxins/immunology , Blood Donors , Humans , Neutralization Tests , Russia , Spores, Bacterial/immunology , VaccinationABSTRACT
BACKGROUND: Diabetes mellitus is frequently associated with microcirculation pathology and hemorheological disorders. METHODS: 24 patients with diabetic foot and 22 healthy subjects were recruited. RBC aggregation, disaggregation and morphology of aggregates were determined in autologous plasma and serum. RESULTS: The RBC aggregation in patients with diabetic foot increased in autologous plasma and serum. Increased red blood cell aggregate strength in these patients was observed only in autologous plasma. Microscopic images of RBC aggregates of patients with diabetic foot show the formation of pathologic globular structures of aggregates in autologous plasma and serum. CONCLUSION: The RBC aggregation in autologous plasma and autologous serum in patients with diabetic foot is significantly higher than in healthy subjects. Increase in strength of RBC aggregates in diabetic foot patients was observed only in autologous plasma. The microscopic images of RBC aggregates in patients with diabetic foot indicate the formation of globular (pathologic) structures of aggregates in autologous plasma and serum. The differences in the morphology of RBC aggregates in autologous plasma and serum between healthy subjects and diabetic foot patients, obtained by microscopic image analysis with high magnification light microscope, can be used as an additional diagnostic tool in medical practice.