Integration of immunoinformatics and cheminformatics to design and evaluate a multitope vaccine against Klebsiella pneumoniae and Pseudomonas aeruginosa coinfection.

Introduction: Klebsiella pneumoniae (K. pneumoniae) and Pseudomonas aeruginosa (P. aeruginosa) are the most common Gram-negative bacteria associated with pneumonia and coinfecting the same patient. Despite their high virulence, there is no effective vaccine against them. Methods: In the current study, the screening of several proteins from both pathogens highlighted FepA and OmpK35 for K. pneumonia in addition to HasR and OprF from P. aeruginosa as promising candidates for epitope mapping. Those four proteins were linked to form a multitope vaccine, that was formulated with a suitable adjuvant, and PADRE peptides to finalize the multitope vaccine construct. The final vaccine's physicochemical features, antigenicity, toxicity, allergenicity, and solubility were evaluated for use in humans. Results: The output of the computational analysis revealed that the designed multitope construct has passed these assessments with satisfactory scores where, as the last stage, we performed a molecular docking study between the potential vaccine construct and K. pneumonia associated immune receptors, TLR4 and TLR2, showing affinitive to both targets with preferentiality for the TLR4 receptor protein. Validation of the docking studies has proceeded through molecular dynamics simulation, which estimated a strong binding and supported the nomination of the designed vaccine as a putative solution for K. pneumoniae and P. aeruginosa coinfection. Here, we describe the approach for the design and assessment of our potential vaccine.

The development of ghost vaccines trials.

INTRODUCTION: Bacterial ghosts are intact bacterial cell envelopes that are emptied of their content by gentle biological or chemical poring methods. Ghost techniques increase the safety of the killed vaccines, while maintaining their antigenicity due to mild preparation procedures. Moreover, ghost-platforms may express and/or carry several antigens or plasmid-DNA encoding for protein epitopes. AREAS COVERED: In this review, the development in ghost-vaccine production over the last 30 years is classified and discussed. The different applications of ghost-vaccines, how they trigger the immune system, their advantages and limitations are displayed. The phage-mediated lysis, molecular manipulation of the lysis-genes, and the biotechnological production of ghosts are described. The trials are classified according to the pattern of lysis and to the type of bacteria. Further subdivision includes chronological ordered application of the ghost as alternative-killed vaccine, recombinant antigen platform, plasmid DNA carrier, adjuvants, and dendritic cell inducer. Particular trials for specific pathogens or from distinct research schools are gathered. EXPERT OPINION: Ghosts are highly qualified to act as immune-presenting platforms that express and/or carry several recombinant and DNA vaccines, as well as, being efficient alternative-killed vaccines. The coming years will show more molecular advances to develop ghost-production and to express more antigens.

The detection of antigenic determinants of Acinetobacter baumannii.

BACKGROUND: Acinetobacter baumannii continues to pose a threat to burdened patients in ICUs all around the world. Lately, infection control techniques are not sufficient to curb A. baumannii's progression and chemotherapeutics are losing their potency against it. Thus, immunization became a key player in providing an ideal solution to the dilemma. None of the vaccines under investigation have reached the market and the search for a tailored vaccine remains a challenge. The notion of unravelling the bacterial antigens to design a novel epitope-based vaccine proved its merits. METHODS: In this work, the propitious polysaccharide and protein antigenic determinants of A. baumannii were mapped by mimicking the infection. The immune response was evaluated by western blot, ELISA, and cellular proliferation assay techniques. RESULTS: The screening showed that OMPs induced the most eminent sustained IgG response. In addition, OMP gave the highest cellular proliferation and a fold increase in ELISA that reached up to 10-fold by week 6. Whilst, the LPS gave a rapid IgM response, that reached 5-fold and the response was visible from week 1 in the western blot. The OMPs had a more pronounced effect in eliciting a cellular immune response. CONCLUSION: The results elaborated the valuable role of using pure OMPs and detoxified LPS together; as a major cornerstone in designing an ideal vaccine against A. baumannii.

Development of immunization trials against Pasteurella multocida.

Pasteurellosis is one of the most important respiratory diseases facing economically valuable farm animals such as poultry, rabbit, cattle, goats and pigs. It causes severe economic loss due to its symptoms that range from primary local infection to fatal septicemia. Pasteurella multocida is the responsible pathogen for this contagious disease. Chemotherapeutic treatment of Pasteurella is expensive, lengthy, and ineffective due to the increasing antibiotics resistance of the bacterium, as well as its toxicity to human consumers. Though, biosecurity measures played a role in diminishing the spread of the pathogen, the immunization methods were always the most potent preventive measures. Since the early 1950s, several trials for constructing and formulating effective vaccines were followed. This up-to-date review classifies and documents such trials. A section is devoted to discussing each group benefits and defects.

The use of propolis as vaccine's adjuvant.

The success of many vaccines relies on their association with selected adjuvants in order to increase their immunogenicity and ensure long-term protection. All available adjuvants have adverse effects due to their toxicity and reactogenicity. Pre-clinical in vivo investigations can identify new natural products for further applications. Several studies have confirmed the different medicinal benefits of propolis. However the studies that addressed its use as a potent, safe, vaccine adjuvant were limited to specific countries and languages, primarily Chinese. Those studies introduced the use of different extracts and formulations of propolis as adjuvants for bacterial, viral, and parasitic vaccines. This comprehensive up-to-date review categorizes, documents, and discusses those trials in a clear chronological manner.

Development of immunization trials against Klebsiella pneumoniae.

Klebsiella pneumoniae is the most common cause of nosocomial respiratory tract and premature intensive care infections, and the second most frequent cause of Gram-negative bacteraemia and urinary tract infections. Drug resistant isolates remain an important hospital-acquired bacterial pathogen, add significantly to hospital stays, and are especially problematic in high impact medical areas such as intensive care units. Many investigations worldwide proved the increasing resistance of such pathogen, resulting in an average rate of 1.63 outbreak every year. A variety of preventive measures were applied to reduce such incidences. Immunotherapy and passive immunization researches as well found their way to the treatment of Klebsiella. During the last 40 years, many trials for constructing effective vaccines were followed. This up-to-date review classifies such trials and documents them in a progressive way. A following comment discusses each group benefits and defects.

Development of a new trend conjugate vaccine for the prevention of Klebsiella pneumoniae.

Klebsiella pneumoniae is a major cause of nosocomial pneumonia, septicemia and urinary tract infections, especially in newborns, blood cancer patients, and other immunocompromised candidates. The control of K. pneumoniae is a complicated issue due to its tight pathogenesis. Immuno-prophylactic preparations, especially those directed toward the bacterium O-antigen, showed to be the most successful way to prevent the infection incidence. However, all previously proposed preparations were either of limited spectrum or non-maternal, and hence not targeting the main Klebsiella patients. Moreover, all preparations were directed only to prevent the respiratory diseases due to that pathogen. This article addresses the development of a method originally used to purify the non-capsular bacterial-endotoxins, as a new and easy method for vaccine production against K. pneumoniae. The application of this method was preceded by a biotechnological control of capsular polysaccharide production in K. pneumoniae. The new produced natural conjugate between the bacterial O-antigen and its outer membrane proteins was evaluated by physicochemical and immunological methods to investigate its purity, integrity, safety and immunogenicity. It showed to be pure, stable, safe for use, and able to elicit a protective immunoglobulin titer against different Klebsiella infections. This immune-response proved to be transferable to the offspring of the vaccinated experimental rabbits via placenta.