Pseudomonas aeruginosa outer-membrane protein F epitopes are highly immunogenic in mice when expressed on a plant virus.
A synthetic peptide (peptide 10) representing a surface-exposed, linear B cell epitope from outer-membrane (OM) protein F of Pseudomonas aeruginosa was shown previously to afford protection in mice from P. aeruginosa infection. This peptide was expressed in tandem with the protein F peptide 18 on each of the two coat proteins of cowpea mosaic virus (CPMV). The chimaeric virus particles (CVPs) expressing the peptides on the S (small) coat protein (CPMV-PAE4) and L (large) coat protein (CPMV-PAE5) were used to immunize mice. Following subcutaneous immunization in Freund's and QuilA adjuvants, CPMV-PAE4 induced antibodies predominantly against peptide 18, whereas CPMV-PAE5 produced antibodies exclusively against peptide 10, indicating that the site of peptide expression on CPMV influences its immune recognition. The anti-peptide antibodies elicited by CPMV-PAE5 were predominantly of the IgG2a isotype, indicating a highly polarized TH1-type response. The peptide-specific IgG2a strongly recognized the whole F protein, but more importantly, recognized protein F in all seven Fisher-Devlin immunotypes of P. aeruginosa. Furthermore, the peptide-specific IgG2a in CVP/QS-21 adjuvant-immunized mice was shown to bind complement and to augment phagocytosis of P. aeruginosa by human neutrophils in vitro. The ability of CPMV-PAE5 to induce P. aeruginosa-specific opsonic IgG2a gives it potential for further development as a protective vaccine against P. aeruginosa.
Brennan
F R
FR
Axis Genetics plc, Babraham, Cambridge CB2 4AZ, UK.
Jones
T D
TD
Axis Genetics plc, Babraham, Cambridge CB2 4AZ, UK.
Gilleland
L B
LB
Department of Microbiology and Immunology, Louisiana State University Medical Center, School of Medicine in Shreveport, Shreveport, LA 71130-3932, USA.
Bellaby
T
T
Axis Genetics plc, Babraham, Cambridge CB2 4AZ, UK.
Xu
F
F
Axis Genetics plc, Babraham, Cambridge CB2 4AZ, UK.
North
P C
PC
Axis Genetics plc, Babraham, Cambridge CB2 4AZ, UK.
Thompson
A
A
Axis Genetics plc, Babraham, Cambridge CB2 4AZ, UK.
Staczek
J
J
Department of Microbiology and Immunology, Louisiana State University Medical Center, School of Medicine in Shreveport, Shreveport, LA 71130-3932, USA.
Lin
T
T
Department of Molecular Biology, The Scripps Research Institute, 10666 North Torrey Pines Road, La Jolla, CA 92037, USA.
Johnson
J E
JE
Department of Molecular Biology, The Scripps Research Institute, 10666 North Torrey Pines Road, La Jolla, CA 92037, USA.
Hamilton
W D O
WDO
Axis Genetics plc, Babraham, Cambridge CB2 4AZ, UK.
Gilleland
H E
HE
Department of Microbiology and Immunology, Louisiana State University Medical Center, School of Medicine in Shreveport, Shreveport, LA 71130-3932, USA.
D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis*; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics; D017798:Comovirus / Q000235:genetics* / Q000276:immunology; D003165:Complement System Proteins / Q000276:immunology; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007114:Immunization; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009504:Neutrophils / Q000276:immunology; D009895:Opsonin Proteins / Q000097:blood / Q000276:immunology; D010587:Phagocytosis; D018272:Porins / Q000235:genetics / Q000276:immunology*; D011552:Pseudomonas Infections / Q000145:classification / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control; D011550:Pseudomonas aeruginosa / Q000276:immunology*; D018417:Th1 Cells / Q000276:immunology; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology
D000907:Antibodies, Bacterial; D000939:Epitopes; D007074:Immunoglobulin G; C109237:OmpF protein; D009895:Opsonin Proteins; D018272:Porins; D014614:Vaccines, Synthetic; D003165:Complement System Proteins