Experimental acute respiratory Burkholderia pseudomallei infection in BALB/c mice.

Burkholderia pseudomallei is the causative agent of melioidosis, which is considered a potential deliberate release agent. The objective of this study was to establish and characterise a relevant, acute respiratory Burkholderia pseudomallei infection in BALB/c mice. Mice were infected with 100 B. pseudomallei strain BRI bacteria by the aerosol route (approximately 20 median lethal doses). Bacterial counts within lung, liver, spleen, brain, kidney and blood over 5 days were determined and histopathological and immunocytochemical profiles were assessed. Bacterial numbers in the lungs reached approximately 10(8) cfu/ml at day 5 post-infection. Bacterial numbers in other tissues were lower, reaching between 10(3) and 10(5) cfu/ml at day 4. Blood counts remained relatively constant at approximately 1.0 x 10(2) cfu/ml. Foci of acute inflammation and necrosis were seen within lungs, liver and spleen. These results suggest that the BALB/c mouse is highly susceptible to B. pseudomallei by the aerosol route and represents a relevant model system of acute human melioidosis.

Experimental aerogenic Burkholderia mallei (glanders) infection in the BALB/c mouse.

The object of this study was to develop and characterize experimental Burkholderia mallei aerosol infection in BALB/c mice. Sixty-five mice were infected with 5000 [approx. 2.5 median lethal doses (MLD)] B. mallei strain ATCC 23344(T) bacteria by the aerosol route. Bacterial counts within lung, liver, spleen, brain, kidney and blood over 14 days were determined and histopathological and immunocytochemical profiles were assessed. Mortality due to B. mallei infection occurred between days 4 and 10 post-infection. Bacterial numbers were consistently higher in the lungs than in other tissues, reaching a maximum of approximately 1.0 x 10(6) c.f.u. ml(-1) at 5 days post-infection. Bacterial counts in liver and spleen tissue remained approximately equal, reaching a maximum of approximately 1.0 x 10(4) c.f.u. ml(-1) at day 4 post-infection. By day 14 post-infection, bacterial counts were in the range 1.0 x 10(3)-1.0 x 10(4) c.f.u. ml(-1) for all tissues. Infection of the lungs by B. mallei resulted in foci of acute inflammation and necrosis. As infection progressed, the inflammatory process became subacute or chronic; this was associated with the development of extensive consolidation. Lesions in liver and spleen tissue were typical of those that might be expected in bacteraemia or bacterial toxaemia. These results suggest that the BALB/c mouse is susceptible to B. mallei when delivered by the aerosol route and that this represents a model system of acute human glanders that is suitable for research into the pathogenesis of and vaccines against this disease.

Mucosal or parenteral administration of microsphere-associated Bacillus anthracis protective antigen protects against anthrax infection in mice.

Existing licensed anthrax vaccines are administered parenterally and require multiple doses to induce protective immunity. This requires trained personnel and is not the optimum route for stimulating a mucosal immune response. Microencapsulation of vaccine antigens offers a number of advantages over traditional vaccine formulations, including stability without refrigeration and the potential for utilizing less invasive routes of administration. Recombinant protective antigen (rPA), the dominant antigen for protection against anthrax infection, was encapsulated in poly-L-lactide 100-kDa microspheres. Alternatively, rPA was loosely attached to the surfaces of microspheres by lyophilization. All of the microspheric formulations were administered to A/J mice with a two-dose schedule by either the intramuscular route, the intranasal route, or a combination of these two routes, and immunogenicity and protective efficacy were assessed. An intramuscular priming immunization followed by either an intramuscular or intranasal boost gave optimum anti-rPA immunoglobulin G titers. Despite differences in rPA-specific antibody titers, all immunized mice survived an injected challenge consisting of 10(3) median lethal doses of Bacillus anthracis STI spores. Immunization with microencapsulated and microsphere-associated formulations of rPA also protected against aerosol challenge with 30 median lethal doses of STI spores. These results show that rPA can be encapsulated and surface bound to polymeric microspheres without impairing its immunogenicity and also that mucosal or parenteral administration of microspheric formulations of rPA efficiently protects mice against both injected and aerosol challenges with B. anthracis spores. Microspheric formulations of rPA could represent the next generation of anthrax vaccines, which could require fewer doses because they are more potent, are less reactogenic than currently available human anthrax vaccines, and could be self-administered without injection.