Immune response in mice following immunization with DNA encoding fragment C of tetanus toxin.
Tetanus toxin is a potent neurotoxin synthesized by Clostridium tetani. Immunization with fragment C protein, the nontoxic C-terminal domain of tetanus toxin, will protect mice against lethal challenge with tetanus toxin. A synthetic gene encoding fragment C (tetC) had previously been shown to express high levels of fragment C in Saccharomyces cerevisiae. A plasmid, pcDNA3/tetC, which encodes the synthetic tetC gene expressed under the control of the human cytomegalovirus major intermediate-early promoter/enhancer region, was constructed. Expression of fragment C was observed in eukaryotic cells growing in vitro following transfection with pcDNA3/tetC. The immune response induced by intramuscular immunization with pure pcDNA3/tetC DNA was evaluated in a murine model. Anti-fragment C serum immunoglobulin and proliferative responses in splenocytes were observed in BALB/c mice following two immunizations with pcDNA3/tetC. The major immunoglobulin G subclass that recognized fragment C was immunoglobulin G2a, and the stimulated splenocytes secreted high levels of gamma interferon. Immunity to tetanus is dependent on the presence of neutralizing serum antibodies against tetanus toxin. Sufficient anti-fragment C serum immunoglobulins were induced by DNA-mediated immunization to protect mice against lethal challenge with tetanus toxin.
Anderson
R
R
Department of Biochemistry, Imperial College of Science, Technology and Medicine, London, United Kingdom.
Gao
X M
XM
Papakonstantinopoulou
A
A
Roberts
M
M
Dougan
G
G
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001483:Base Sequence; D016207:Cytokines / Q000378:metabolism; D004269:DNA, Bacterial / Q000276:immunology / Q000627:therapeutic use*; D005260:Female; D005822:Genetic Vectors; D007111:Immunity, Cellular; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013742:Tetanus / Q000517:prevention & control*; D013744:Tetanus Toxin / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D014611:Vaccination
D000907:Antibodies, Bacterial; D016207:Cytokines; D004269:DNA, Bacterial; D010446:Peptide Fragments; D013744:Tetanus Toxin; C040635:tetanus toxin fragment C