Selection of an immunogenic and protective epitope of the PsaA protein of Streptococcus pneumoniae using a phage display library.
Streptococcus pneumoniae is an important pathogen that causes disease in young and elderly individuals. The currently available polysaccharide vaccines have limited efficacy in those age groups most susceptible to pneumococcal infections. This study focuses on mapping the epitopes of a surface protein of S. pneumoniae by biopanning a 15 mer phage display library using 5 different monoclonal antibodies (MAbs) against the Pneumoccal surface adhesin A (PsaA). PsaA is a component of the bacterial cell wall that is highly species specific and is involved in bacterial adherence and virulence. Biopanning of the phage display library reveals three distinct epitopes on the PsaA protein. The sequence homology of these epitopes ranges from two to six amino acids when compared to the native PsaA protein type 2. Two of these epitopes have been evaluated for their immunogeneicity in mice. The peptide selected by the MAbs 8G12, 6F6, and 1B7 is referred to as the consensus peptide and is immunogenic in mice. Optimal anti-PsaA response is observed in mice immunized with 50microg of the consensus peptide complexed to proteosomes in 1:1 ratio. The anti-PsaA response is significantly lower than the response to the PsaA native protein. The peptide selected by monoclonal antibody 4E9 in its lipidated form is significantly protective in mice challenged with S. pneumoniae serotype 2 when compared to mice immunized with the native protein. These results show that the selected epitopes of PsaA protein are immunogenic and protective in mice. These epitopes need to be evaluated further as alternatives to currently available vaccines.
Srivastava
N
N
Department of Medicine, Medical College of Ohio, Toledo 43614, USA.
Zeiler
J L
JL
Smithson
S L
SL
Carlone
G M
GM
Ades
E W
EW
Sampson
J S
JS
Johnson
S E
SE
Kieber-Emmons
T
T
Westerink
M A
MA
D018829:Adhesins, Bacterial; D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D001426:Bacterial Proteins / Q000008:administration & dosage / Q000032:analysis* / Q000276:immunology; D002352:Carrier Proteins / Q000008:administration & dosage / Q000032:analysis* / Q000276:immunology; D016056:Immunodominant Epitopes / Q000008:administration & dosage / Q000032:analysis* / Q000276:immunology; D008074:Lipoproteins / Q000008:administration & dosage / Q000032:analysis* / Q000276:immunology; D026901:Membrane Transport Proteins; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D019151:Peptide Library; D011008:Pneumococcal Infections / Q000276:immunology / Q000517:prevention & control*; D017108:Streptococcus Phages / Q000235:genetics / Q000276:immunology; D013296:Streptococcus pneumoniae / Q000276:immunology* / Q000821:virology
D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D002352:Carrier Proteins; D016056:Immunodominant Epitopes; D008074:Lipoproteins; D026901:Membrane Transport Proteins; D019151:Peptide Library; C085000:PsaA protein, Streptococcus