ABSTRACT
An extensive morphological analysis of the Neisseria meningitidis cell envelope, including serogroup B capsule and outer membrane, based on atomic force microscopy (AFM) together with mechanical characterization by force spectroscopic measurements, has been carried out. Three meningococcal strains were used: the encapsulated serogroup B strain B1940, and the isogenic mutants B1940 siaD(+C) (lacking capsule), and B1940 cps (lacking both capsule and lipooligosaccharide outer core). AFM experiments with the encapsulated strain B1940 provided unprecedented images of the meningococcal capsule, which seems to be characterized by protrusions ("bumps") with the lateral dimensions of about 30 nm. Measurement of the Young's modulus provided quantitative assessment of the property of the capsule to confer resistance to mechanical stress. Moreover, Raman spectroscopy gave a fingerprint by which it was possible to identify the specific molecular species of the three strains analyzed, and to highlight major differences between them.
Subject(s)
Bacterial Capsules/ultrastructure , Bacterial Outer Membrane/ultrastructure , Neisseria meningitidis, Serogroup B/ultrastructure , Bacterial Capsules/chemistry , Bacterial Capsules/physiology , Bacterial Outer Membrane/chemistry , Bacterial Outer Membrane/physiology , Elastic Modulus , Microscopy, Atomic Force , Neisseria meningitidis, Serogroup B/chemistry , Neisseria meningitidis, Serogroup B/genetics , Polysaccharides, Bacterial/chemistry , Spectrum Analysis, Raman , Stress, Mechanical , Surface PropertiesABSTRACT
Meningitis and sepsis caused by serogroup B meningococcus are two severe diseases that still cause significant mortality. To date there is no universal vaccine that prevents these diseases. In this work, five antigens discovered by reverse vaccinology were expressed in a form suitable for large-scale manufacturing and formulated with adjuvants suitable for human use. The vaccine adjuvanted by aluminum hydroxide induced bactericidal antibodies in mice against 78% of a panel of 85 meningococcal strains representative of the global population diversity. The strain coverage could be increased to 90% and above by the addition of CpG oligonucleotides or by using MF59 as adjuvant. The vaccine has the potential to conquer one of the most devastating diseases of childhood.