Expression and immune response to foreign epitopes in bacteria. Perspectives for live vaccine development.
We previously developed a general procedure which allows the genetic coupling of a chosen foreign linear epitope in different regions of a carrier protein. By using as carriers, two bacterial envelope proteins, the LamB and MalE proteins of E. coli K12, we were able to express the same epitope in different sites of the two proteins and in different compartments of the bacteria. This allowed us to analyze the influence of the localization in E. coli cells of a foreign B-cell epitope on the induction of specific antibody responses, and the role of the molecular environment on the immunological properties of foreign B- or T-cell epitopes, using either purified hybrid proteins or live recombinant bacteria. Several LamB and MalE hybrid proteins were expressed in the aroA attenuated strain of S. typhimurium, SL3261. Immunizations of mice with live recombinant bacteria by the intravenous route showed that it was possible to induce humoral responses against inserted foreign sequences. In order to improve the in vivo stability of the plasmids carrying the different contructions, and to increase the amounts of recombinant LamB and MalE hybrid proteins expressed in vivo, the LamB and malE genes were placed under the control of the anaerobically inducible pnirBpromoter control. The genetic factors susceptible of influencing the immune response to recombinant Salmonella in mice were also studied.
Charbit
A
A
Unité de Programmation Moléculaire et Toxicologie Génétique, Institut Pasteur, Paris, France.
Newton
S M
SM
Klebba
P E
PE
Clément
J M
JM
Fayolle
C
C
Lo-Man
R
R
Leclerc
C
C
Hofnung
M
M
D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D018528:ATP-Binding Cassette Transporters; D019883:Alkyl and Aryl Transferases / Q000096:biosynthesis / Q000276:immunology; D000818:Animals; D000917:Antibody Formation; D001402:B-Lymphocytes / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D002352:Carrier Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D003001:Cloning, Molecular; D015195:Drug Design; D000939:Epitopes / Q000096:biosynthesis / Q000276:immunology*; D004926:Escherichia coli / Q000276:immunology*; D029968:Escherichia coli Proteins; D058619:Maltose-Binding Proteins; D051379:Mice; D008961:Models, Structural; D009004:Monosaccharide Transport Proteins; D033902:Periplasmic Binding Proteins; D018272:Porins; D011401:Promoter Regions, Genetic; D017433:Protein Structure, Secondary; D011991:Receptors, Virus / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology; D012486:Salmonella typhimurium / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic
D018528:ATP-Binding Cassette Transporters; D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D002352:Carrier Proteins; D000939:Epitopes; D029968:Escherichia coli Proteins; C461619:MalE protein, E coli; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D033902:Periplasmic Binding Proteins; D018272:Porins; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic; C016532:maltoporins; C461599:maltose transport system, E coli; D019883:Alkyl and Aryl Transferases; D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase