The present invention relates to, in part, compositions comprising improved flagellin derived constructs and methods of using for vaccination, including adjuvants comprising flagellin-based agents.

The present invention relates to the fields of medical microbiology and vaccines. In particular the invention relates to a process wherein the spontaneous release of bacterial outer membrane vesicles (OMV) of Gram-negative bacteria is stimulated by application of a dissolved oxygen tension (DOT) that is higher than a physiological DOT. The thus produced OMVs are for use in vaccines. The invention further relates to OMV obtainable by said process, and to a pharmaceutical composition comprising such OMV. The present invention further relates to the use of OMV of the present invention as a medicament in particular for use in a method for eliciting an immune response.

Disclosed are methods, systems, components, and compositions for cell-free synthesis of glycosylated proteins. The glycosylated proteins may be utilized in vaccines, including anti-bacterial vaccines. The glycosylated proteins may include a bacterial polysaccharide conjugated to a carrier, which may be utilized to generate an immune response in an immunized host against the polysaccharide conjugated to the carrier. The glycosylated proteins may be synthesized in cell-free glycoprotein synthesis (CFGpS) systems using prokaryote cell lysates that are enriched in components for glycoprotein synthesis such as oligosaccharyltransferases (OSTs) and lipid-linked oligosaccharides (LLOs) including OSTs and LLOs associated with synthesis of bacterial O antigens.

Targeted disruption of a specific gene and its subsequent restoration in obligate intracellular bacteria remains extremely challenging due to their absolute requirement for residence inside a host cell to replicate. Here, targeted allelic exchange mutations were created to inactivate two genes and then to restore one of the two genes of a rickettsial pathogen, Ehrlichia chaffeensis. These methods were then also successfully utilized in Ehrlichia canis and Anaplasma phagocyophilum. The resultant mutated pathogens are useful in immunogenic compositions for reducing the incidence of or severity of infection with ricksettsial pathogens.

The present invention relates to a method of preparing a live attenuated vaccine by irradiation and a live attenuated vaccine composition prepared by the same, and more particularly, a method of preparing a live attenuated vaccine by irradiation including irradiating a pathogenic microorganism with a dose of 0.5 to 2 kGy of radiation per single radiation six to fifteen times; and a live attenuated vaccine composition including a pathogenic microorganism attenuated to not be revertant to a wild type by generation of at least one mutation of nucleotide insertion and nucleotide deletion by irradiation.

The present invention relates to a method of preparing a live attenuated vaccine by irradiation and a live attenuated vaccine composition prepared by the same, and more particularly, a method of preparing a live attenuated vaccine by irradiation including irradiating a pathogenic microorganism with a dose of 0.5 to 2 kGy of radiation per single radiation six to fifteen times; and a live attenuated vaccine composition including a pathogenic microorganism attenuated to not be revertant to a wild type by generation of at least one mutation of nucleotide insertion and nucleotide deletion by irradiation.

A system including a vial and methods of its use to provide a lyophilized cake removable from the vial without cracking or breaking.

In one aspect, the invention relates to an immunogenic composition that includes a mutant Clostridium difficile toxin A and/or a mutant Clostridium difficile toxin B. Each mutant toxin includes a glucosyltransferase domain having at least one mutation and a cysteine protease domain having at least one mutation, relative to the corresponding wild-type C. difficile toxin. The mutant toxins may further include at least one amino acid that is chemically crosslinked. In another aspect, the invention relates to antibodies or binding fragments thereof that binds to said immunogenic compositions. In further aspects, the invention relates to isolated nucleotide sequences that encode any of the foregoing, and methods of use of any of the foregoing compositions.

Outer membrane vesicles from bacteria of the Burkholderia pseudomallei complex can be used as adjuvants in compositions and methods to potentiate the immune response to immunogens.

Outer membrane vesicles from bacteria of the Burkholderia pseudomallei complex can be used as adjuvants in compositions and methods to potentiate the immune response to immunogens.