Optimization of the Process for Preparing Bivalent Polysaccharide Conjugates to Develop Multivalent Conjugate Vaccines against Streptococcus pneumoniae or Neisseria meningitidis and Comparison with the Corresponding Licensed Vaccines in Animal Models.

OBJECTIVE: This study aimed to describe, optimize and evaluate a method for preparing multivalent conjugate vaccines by simultaneous conjugation of two different bacterial capsular polysaccharides (CPs) with tetanus toxoid (TT) as bivalent conjugates. METHODS: Different molecular weights (MWs) of polysaccharides, activating agents and capsular polysaccharide/protein (CP/Pro) ratio that may influence conjugation and immunogenicity were investigated and optimized to prepare the bivalent conjugate bulk. Using the described method and optimized parameters, a 20-valent pneumococcal conjugate vaccine and a bivalent meningococcal vaccine were developed and their effectiveness was compared to that of corresponding licensed vaccines in rabbit or mouse models. RESULTS: The immunogenicity test revealed that polysaccharides with lower MWs were better for Pn1-TT-Pn3 and MenA-TT-MenC, while higher MWs were superior for Pn4-TT-Pn14, Pn6A-TT-Pn6B, Pn7F-TT-Pn23F and Pn8-TT-Pn11A. For activating polysaccharides, 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) was superior to cyanogen bromide (CNBr), but for Pn1, Pn3 and MenC, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDAC) was the most suitable option. For Pn6A-TT-Pn6B and Pn8-TT-Pn11A, rabbits immunized with bivalent conjugates with lower CP/Pro ratios showed significantly stronger CP-specific antibody responses, while for Pn4-TT-Pn14, higher CP/Pro ratio was better. Instead of interfering with the respective immunological activity, our bivalent conjugates usually induced higher IgG titers than their monovalent counterparts. CONCLUSION: The result indicated that the described conjugation technique was feasible and efficacious to prepare glycoconjugate vaccines, laying a solid foundation for developing extended-valent multivalent or combined conjugate vaccines without potentially decreased immune function.

Isolation and characterization of heavy-metal-mobilizing bacteria from contaminated soils and their potential in promoting Pb, Cu, and Cd accumulation by Coprinus comatus.

The enhanced effect of heavy-metal-mobilizing bacteria on the uptake of Pb, Cu, and Cd by Coprinus comatus from Pb-, Cu-, and Cd-multicontaminated soil was assessed in this study. Thirteen strains, tolerating 800 mg·L(-1) Pb, 200 mg·L(-1) Cu, and 200 mg·L(-1) Cd simultaneously were selected for heavy-metal-solubilizing experiments in soil. The mobilization of heavy metals depended on the characteristics of bacteria and heavy metals. Correlation analysis demonstrated that for Pb solubilization, the acid-producing ability was the most significant factor, while for Cu and Cd, siderophores played a leading role in this process. Four strains, based on their excellent ability to solubilize heavy metal in soil, were applied in pot experiments. The results showed that all strains can promote the growth of C. comatus and meanwhile help mushroom accumulate more heavy metals (Pb, Cd, and Cu). The maximum uptake for total Pb and Cu by C. comatus was observed in inoculations with Bacillus sp. strain JSG1 (2.02- and 2.13-fold, respectively, compared with uninoculated soil), while for Cd, it was recorded in Bacillus sp. strain PB2 treated soil (2.03-fold). Therefore, this work suggests that the mushroom-bacteria interaction can be developed into a novel bioremediation strategy.