Expression of human IL-1 beta in Salmonella typhimurium. A model system for the delivery of recombinant therapeutic proteins in vivo.
The feasibility of using Salmonella typhimurium aroA mutant (SL3261) to deliver protein therapeutic agents was investigated in a murine model system. We have constructed an Escherichia coli expression plasmid designed to express the human protein IL-1 beta. This plasmid expresses IL-1 beta to high levels (greater than 30% total cell protein) in E. coli. In Salmonella the IL-1 beta is expressed constitutively to about 10% total cell protein, as verified by Western blotting analysis using polyclonal rabbit anti-IL-1 beta antibody. The protein is produced in a soluble and biologically active form. BALB/c mice administered orally or i.v. with S. typhimurium aroA mutants carrying the plasmid produced highly significant antibody responses against human IL-1 beta as determined by a solid-phase RIA. Furthermore, mice injected with the construct were significantly protected against lethal gamma-irradiation (850 rad). This study therefore demonstrates that the vaccine strain of Salmonella mutants can also be used effectively to deliver therapeutic proteins in vivo.
Carrier
M J
MJ
Wellcome Research Laboratories, Beckenham, Kent, England.
Chatfield
S N
SN
Dougan
G
G
Nowicka
U T
UT
O'Callaghan
D
D
Beesley
J E
JE
Milano
S
S
Cillari
E
E
Liew
F Y
FY
D000818:Animals; D001428:Bacterial Vaccines; D016503:Drug Delivery Systems; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D014677:Pharmaceutical Vehicles; D010957:Plasmids; D011994:Recombinant Proteins / Q000008:administration & dosage*; D012486:Salmonella typhimurium / Q000235:genetics / Q000378:metabolism*; D014916:Whole-Body Irradiation
D001428:Bacterial Vaccines; D007375:Interleukin-1; D014677:Pharmaceutical Vehicles; D011994:Recombinant Proteins