Genetic immunisation against hepatitis B using whole bacteriophage λ particles

Phage Library Screening for the Rapid Identification and In Vivo Testing of Candidate Genes for a DNA Vaccine against Mycoplasma mycoides subsp. mycoides Small Colony Biotype

“…IPTG (0.2 mM) was then added, and incubation was continued for a further 3 to 4 h. Cells were pelleted and resuspended in a small volume (100 to 400 l, depending on the cell density) of SDS-PAGE sample buffer (0.1 M Tris-HCl, pH 6.8, 40% [vol/vol] glycerol, 4% [wt/vol] SDS, 0.25% bromophenol blue, 2% ␤-mercaptoethanol). The samples were boiled for 5 min, separated by SDS-PAGE, and immunoblotted using standard procedures as previously described (20). Blots were probed using rabbit (R55) or mouse primary antiserum and a relevant secondary antiserum (DAKO A/S, Glostrup, Denmark).…”

“…mycoides SC biotype clones were grown on strain XL1-Blue (Stratagene). Large-scale liquid lysates of phage were prepared, purified using previously published protocols (essentially polyethylene glycol precipitation followed by ultracentrifugation) (20), and stored at 4°C until use.…”

“…After challenge, mice vaccinated with clone -A8 also exhibited a reduced level of mycoplasmemia compared to controls, suggesting that the MSC_0397 lipoprotein has a protective effect in the mouse model when delivered as a bacteriophage DNA vaccine. Bacteriophage-mediated immunoscreening using an appropriate vector system offers a rapid and simple technique for the identification and immediate testing of putative candidate vaccines from a variety of pathogens.Whole bacteriophage (phage) particles have recently been described as highly efficient DNA vaccine delivery vehicles (6,7,17,20), inducing significant antibody responses in both laboratory mice (7) and larger animals such as rabbits (20) and sheep (33). Bacteriophages are viruses of bacteria and are metabolically inert, requiring the host bacterium for growth and propagation.…”

“…Vaccination with phage particles also induces a highly immunogenic signal (against phage coat proteins), which provides an easily assayable marker to confirm the vaccination status of animals. Previous research has indicated that high phage antibody titers do not interfere with the immune response against the expressed DNA vaccine antigen, and if anything, are more likely to efficiently target the phage to APCs (via phage-antibody complexes [20]). …”

“…Whole bacteriophage (phage) particles have recently been described as highly efficient DNA vaccine delivery vehicles (6,7,17,20), inducing significant antibody responses in both laboratory mice (7) and larger animals such as rabbits (20) and sheep (33). Bacteriophages are viruses of bacteria and are metabolically inert, requiring the host bacterium for growth and propagation.…”