lipid-a and Anthrax

Subunit vaccines are theoretically safe and easy to manufacture but require effective adjuvants and delivery systems to yield protective immunity, particularly at critical mucosal sites such as the lung. We investigated nanolipoprotein particles (NLPs) containing the Toll-like receptor 4 agonist monophosphoryl lipid A (MPLA) as a platform for intranasal vaccination against

Topics: Adjuvants, Immunologic; Administration, Intranasal; Animals; Anthrax; Anthrax Vaccines; Antibodies, Bacterial; Bacillus anthracis; Female; Lipid A; Mice; Mice, Inbred BALB C; Nanoparticles; Spores, Bacterial; Vaccines, Subunit

An intranasal vaccine targeting the Bacillus anthracis toxin and vegetative bacterium was tested for the ability to protect immunized rabbits against aerosol B. anthracis spore exposure. Rabbits were vaccinated intranasally with PA-based vaccines formulated as dry powders with or without chitosan (ChiSys, Archimedes Development Limited), a compound that exhibits muco-adhesive properties, or as a liquid. Formulations also contained MPL adjuvant and PA. Some vaccines contained PA conjugated to a 10-mer peptide of the poly-d-glutamic acid capsule of B. anthracis. Rabbits were immunized on days 0 and 28 and aerosol challenged with an average 250LD50 Ames spores on day 85. Serum antibody was measured before and after challenge. Significant anti-PA serum IgG levels were obtained, particularly with use of ChiSys based formulations. PA-Conj induced significant anti-capsule responses, although a formulation containing free capsule peptide did not. All immunized rabbits survived the challenge, but differences in morbidity, as evidenced by anorexia, between vaccine groups were observed. Only rabbits immunized with PA+PA-Conj appeared normal throughout the post-challenge observation period (14 days), while all that received PA with the free capsule peptide appeared ill at times as evidenced by a failure to eat normally. One negative control rabbit received a lower inhaled spore dose (183LD50) and survived the challenge, although it was anorexic post-challenge. It also had a high level of anti-LF antibodies in its convalescent serum (5400 U/ml), indicating an extensive infection. In contrast, 75% of the immunized rabbits had no LF-specific antibody in their post-challenge sera, and the rest had low levels (< or = 138 U/ml), indicating that infections resulting in toxin production were avoided or greatly reduced. Thus, intranasal immunization with a chitosan-based powder vaccine combining PA and capsule epitopes provided superior protection against B. anthracis infection compared to a single antigen (PA) vaccine, as evidenced by a reduction in morbidity and prevention of death.

Topics: Adjuvants, Immunologic; Administration, Intranasal; Aerosols; Animals; Anorexia; Anthrax; Anthrax Vaccines; Antibodies, Bacterial; Antigens, Bacterial; Bacillus anthracis; Bacterial Capsules; Bacterial Toxins; Chitosan; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Immunoglobulin G; Inhalation Exposure; Lipid A; Neutralization Tests; Polyglutamic Acid; Rabbits; Recombinant Fusion Proteins

The efficacy of recombinant Bacillus anthracis Protective Antigen (rPA) produced in Bacillus subtilis and formulated in Alhydrogel or MPL-TDM-CWS (Ribi adjuvant) has been tested and compared to the licensed UK human vaccine in guinea pigs challenged by the aerosol route with the Ames strain of B. anthracis. rPA combined with the Ribi adjuvant was found to be the only formulation to provide 100% protection from challenge. Analysis of immunological parameters in the individual animals revealed significant differences between the rPA/Ribi vaccine group and rPA/Alhydrogel and human vaccine groups for antigen specific lymphocyte proliferation, PA neutralisation and antigen specific IgG2 levels, but indicated no significant differences in PA-specific IgG1 levels. rPA formulated in Alhydrogel induced a mainly IgG1 response whilst the rPA/Ribi vaccine produced a predominantly IgG2 response.

Topics: Adjuvants, Immunologic; Aerosols; Aluminum Hydroxide; Animals; Anthrax; Antibodies, Bacterial; Bacillus anthracis; Cell Wall Skeleton; Cord Factors; Evaluation Studies as Topic; Female; Guinea Pigs; Immunity, Cellular; Immunoglobulin G; Lipid A; Lung; Lymphocyte Activation; Macrophage Activation; Male; Spores, Bacterial