ABSTRACT
AIM: Category A classified Bacillus anthracis is highly fatal pathogen that causes anthrax and creates challenges for global security and public health. In this study, development of a safe and ideal next-generation subunit anthrax vaccine has been evaluated in mouse model. METHOD AND RESULTS: Protective antigen (PA) and BA3338, a surface layer homology (SLH) domain possessing protein were cloned, expressed in heterologous system and purified by IMAC. Recombinant PA and BA3338 with alum were administered in mouse alone or in combination. The humoral and cell-mediated immune response was measured by ELISA and vaccinated animals were challenged with B. anthracis spores via intraperitoneal route. The circulating IgG antibody titre of anti-PA and anti-BA3338 was found significantly high in the first and second booster sera. A significant enhanced level of IL-4, IFN-γ and IL-12 was observed in antigens stimulated supernatant of splenocytes of PA + BA3338 vaccinated animals. A combination of PA and BA3338 provided 80% protection against 20 LD50 lethal dose of B. anthracis spores. CONCLUSION: Both antigens induced admirable humoral and cellular immune response as well as protective efficacy against B. anthracis spores. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has been evaluated for the first time using BA3338 as a vaccine candidate alone or in combination with well-known anthrax vaccine candidate PA. The findings of this study demonstrated that BA3338 could be a co-vaccine candidate for development of dual subunit vaccine against anthrax.
Subject(s)
Anthrax Vaccines/administration & dosage , Anthrax/prevention & control , Antigens, Bacterial/immunology , Bacillus anthracis/immunology , Bacterial Toxins/immunology , Membrane Glycoproteins/immunology , Adjuvants, Immunologic/administration & dosage , Alum Compounds/administration & dosage , Animals , Anthrax/immunology , Anthrax Vaccines/immunology , Antibodies, Bacterial/blood , Cytokines/metabolism , Disease Models, Animal , Immunization/methods , Mice , Vaccines, Subunit/administration & dosage , Vaccines, Subunit/immunologyABSTRACT
Edema factor (EF) is one of the major secretory proteins of anthrax bacteria along with protective antigen (PA) and lethal factor (LF). Edema factor is a calmodulin-and calcium-dependent adenylate cyclase that increases intracellular levels of cAMP. Intracellular trafficking of EF occurs through PA by binding to ATR/CMG2 receptors, which are also involved in other physiological functions of cells. cAMP is a secondary messenger which activates multiple signaling cascades involved in the cytokinetics of actin molecules and cell junction formation. The present study evaluated the effect of EF on growth and angiogenesis patterns in chicken embryos in the in ovo model. Angiogenesis in the chorioallantoic membrane (CAM) of an embryonated chicken egg was decreased and embryo growth was delayed by EF despite the absence of trafficking moiety PA, which is required for transferring the EF molecule inside the cell. Angiogenesis inhibition and embryo growth retardation indicate the use of an alternative receptor by EF to modulate these cellular functions. Additionally, docking was performed between EF as a ligand and hepatocyte growth factor receptor (cMET) and vascular endothelial growth factor (VEGF) receptors, which are mainly involved in growth and angiogenesis. The analysis revealed a very strong binding of EF to cMET receptor (in terms of the number of hydrogen bonds and energy) compared to its ligand hepatocyte growth factor (HGF), which indicates the use of cMET receptor by EF and induction of angiogenesis and embryo growth retardation possibly by competitive inhibition of HGF ligand or receptor-mediated endocytosis.
Subject(s)
Antigens, Bacterial , Bacillus anthracis , Bacterial Toxins , Receptors, Peptide , Adenylyl Cyclases/metabolism , Animals , Antigens, Bacterial/chemistry , Antigens, Bacterial/metabolism , Bacillus anthracis/metabolism , Bacterial Toxins/metabolism , Chick Embryo , Receptors, Peptide/chemistry , Vascular Endothelial Growth Factor A/metabolismABSTRACT
Wound infections are among public health problems worldwide. However, progress has been made in improving surgical techniques and antibiotic treatments. Misuse/overuse of antibiotics to prevent and treat bacterial infections eventually leads to increased bacterial resistance with rising incidences of multi-drug resistant (MDR) bacterial strains. The wider dissemination of antibiotics may ultimately result in ineffectiveness to antibiotic therapy, thereby complicating/graving the outcome of a patient. In the present study, a 60-year-old male patient having wound infection with MDR bacterium that ultimately required surgical amputation of the toe was investigated. For the confirmation of MDR bacterium, two culture media viz., MacConkeyAgar and Mueller Hinton Agar media were used. The sensitivity of the isolated strain for various antibiotics was tested using the disc diffusion method. The wound sample was found positive for Gram-positive bacterium that was identified as Clostridium Perfringens. The bacterium was screened for 40 antibiotics, and among all the antibiotics, it was found sensitive for only Piperacillin/Tazobactam antibiotic combination. C. perfringens bacterium caused the gas gangrene in the infected wound part of the patient. Amputation of the gangrene -affected foot part was performed by surgery, and with good medical care, the person recovered fast. To the best of our knowledge, this is the first-ever report of MDR C. perfringens single isolate harboring resistance against at least 40 antibiotics tested. More research is needed to develop really new and effective medicines that do not cross-react with antibiotics now in use and have robust activity against MDR organisms.
Subject(s)
Clostridium Infections , Clostridium perfringens , Wound Infection , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Clostridium Infections/drug therapy , Drug Resistance, Multiple, Bacterial , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Wound Infection/drug therapy , Wound Infection/microbiologyABSTRACT
OBJECTIVE: Cardiovascular disease (CVD) is the most common non-communicable disease and the leading cause of death worldwide. To reduce the global burden of CVD and related morbidity and mortality, early prediction of CVD risk is essential. Various tools are available to access the risk of cardiovascular disorders. In the present study, we evaluated four risk score calculators associated to CVD for superiority and most reliable CVD prognosis parameters. PATIENTS AND METHODS: In the present prospective study, we investigated the probability of CVD in 150 individuals, including both men and women, using four different cardiovascular risk assessment estimators (Framingham Risk Score [FRS] Calculator, Q-RISK calculator, Reynolds score calculator, and atherosclerotic cardiovascular disease (ASCVD) risk calculator) and evaluated how closely they were related to 16 selected parameters. The four risk estimators shared several common parameters, such as age, smoking status, and blood pressure; however, each of them also used some unique parameters. We used statistical analysis to reduce the number of parameters necessary to predict CVD. RESULTS: Statistical analysis revealed a significant correlation between the main factors responsible for CVD risk. The analysis revealed that out of the four risk calculators tested, the FRS calculator was superior to the others because it showed more significant corroboration with statistical tools and could better predict the most important prognostic factors in CVD. CONCLUSIONS: In all four risk estimators, the parameters that affected risk most significantly and conferred the most reliable CVD prognosis were age, weight, total cholesterol, and hemoglobin levels. With that FRS calculator was superior to the others.