Selection of an Immunogenic and Protective Epitope of the PsaA Protein ofStreptococcus pneumoniaeUsing a Phage Display Library

Abstract

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.

Preparation and immunogenicity of capsular polysaccharide–surface adhesin A (PsaA) conjugate of Streptococcus pneumoniae

“…The studies on the immunity of rPsaA as an antigen were reported (Sandra et al 2006;Srivastava et al 2000). A modest protection against lethal pneumococcal infection was also observed in this work.…”

Differential PsaA-, PspA-, PspC-, and PdB-Specific Immune Responses in a Mouse Model of Pneumococcal Carriage

“…PsaA is highly immunogenic in young children and in the elderly (18). Accordingly, PsaA is a hydrophobic and (palmitoylated) lipoprotein which elicits inherently higher plasma antibody responses than PspA and PdB (43). It is important to mention that PspA-and PspC-specific serum IgG responses during pneumococcal nasal colonization in humans are higher than those directed against PsaA and PdB (31).…”

A Cryptococcal Capsular Polysaccharide Mimotope Prolongs the Survival of Mice withCryptococcus neoformansInfection

“…Recently, there has been increased interest in the use of the random peptide phage display technology to identify peptide mimics of unknown carbohydrate Ags as potential vaccine candidates (24,25,27,(37)(38)(39)(40)(41)(42). In light of the fact that the cryptococcal capsular polysaccharide GXM is poorly immunogenic (43) and defined GXM oligosaccharides are not available, the rationale for the peptide-based approach to vaccine design for C. neoformans was 2-fold: 1) peptide surrogates of polysaccharide Ags selected by defined Abs may direct the response to the production of protective Abs and 2) a peptide-based vaccine should replace the T-independent response to capsular polysaccharide that does not stimulate T cell help (13), with a T-dependent response that induces an amnestic response (40).…”