Protection of immunocompromised mice against lethal infection with Pseudomonas aeruginosa by active or passive immunization with recombinant P. aeruginosa outer membrane protein F and outer membrane protein I fusion proteins.
Recombinant outer membrane proteins (Oprs) of Pseudomonas aeruginosa were expressed in Escherichia coli as glutathione S-transferase (GST)-linked fusion proteins. GST-linked Oprs F and I (GST-OprF190-350 [GST linked to OprF spanning amino acids 190 to 350] and GST-OprI21-83, respectively) and recombinant hybrid Oprs (GST-OprF190-342-OprI21-83 and GST-OprI21-83-OprF190-350) were isolated and tested for their efficacy as vaccines in immunodeficient mice. GST-OprF-OprI protected the mice against a 975-fold 50% lethal dose of P. aeruginosa. Expression of GST-unfused OprF-OprI failed in E. coli, although this hybrid protein has been expressed without a fusion part in Saccharomyces cerevisiae and used for immunizing rabbits. The immune rabbit sera protected severe combined deficient (SCID) mice against a 1,000-fold 50% lethal dose of P. aeruginosa. Evidence is provided to show that the most C-terminal part of OprF (i.e., amino acids 332 to 350) carries an important protective epitope. Opr-based hybrid proteins may have implications for a clinical vaccine against P. aeruginosa.
von Specht
B U
BU
Chirurgische Universitätsklinik, Chirurgische Forschung, Freiburg, Germany.
Knapp
B
B
Muth
G
G
Bröker
M
M
Hungerer
K D
KD
Diehl
K D
KD
Massarrat
K
K
Seemann
A
A
Domdey
H
H
D000818:Animals; D000907:Antibodies, Bacterial / Q000627:therapeutic use*; D000911:Antibodies, Monoclonal / Q000627:therapeutic use; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D001483:Base Sequence; D018604:Epitope Mapping; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D007165:Immunosuppression Therapy; D007167:Immunotherapy; D016233:Immunotherapy, Active; D016219:Immunotherapy, Adoptive; D008074:Lipoproteins / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D051379:Mice; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D008969:Molecular Sequence Data; D018272:Porins / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D011552:Pseudomonas Infections / Q000401:mortality / Q000517:prevention & control*; D011993:Recombinant Fusion Proteins / Q000627:therapeutic use; D012441:Saccharomyces cerevisiae / Q000235:genetics; D016019:Survival Analysis
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D008074:Lipoproteins; D018272:Porins; D011993:Recombinant Fusion Proteins; C060507:outer membrane lipoprotein I, Bacteria