ABSTRACT
Pneumococcal surface adhesin A (PsaA) is a surface-exposed common 37-kilodalton multi-functional lipoprotein detected on all known serotypes of Streptococcus pneumoniae. This lipoprotein belongs to the ABC-type transport protein complex that transports Mn(2+); it is also an adhesin that plays a major role in pneumococcal attachment to the host cell and virulence. PsaA is immunogenic and natural nasopharyngeal colonization of pneumococci elicits an increase in antibody towards PsaA. Hence, PsaA is being actively evaluated as a component of a vaccine in formulations composed of pneumococcal common proteins. PsaA has been expressed as an E. coli recombinant protein, purified, and evaluated in a phase one clinical trial. This article reviews PsaA, its structure and role in pneumococcal virulence, immunogenicity, and potential to reduce nasopharyngeal colonization (a major prerequisite for pneumococcal pathogenesis) as a component of a common pneumococcal protein vaccine.
ABSTRACT
Pneumococcal surface adhesin A (PsaA) is a surface-exposed common 37-kilodalton multi-functional lipoprotein detected on all known serotypes of Streptococcus pneumoniae. This lipoprotein belongs to the ABC-type transport protein complex that transports Mn2+; it is also an adhesin that plays a major role in pneumococcal attachment to the host cell and virulence. PsaA is immunogenic and natural nasopharyngeal colonization of pneumococci elicits an increase in antibody towards PsaA. Hence, PsaA is being actively evaluated as a component of a vaccine in formulations composed of pneumococcal common proteins. PsaA has been expressed as an E. coli recombinant protein, purified, and evaluated in a phase one clinical trial. This article reviews PsaA, its structure and role in pneumococcal virulence, immunogenicity, and potential to reduce nasopharyngeal colonization (a major prerequisite for pneumococcal pathogenesis) as a component of a common pneumococcal protein vaccine.
Subject(s)
Adhesins, Bacterial/immunology , Adhesins, Bacterial/physiology , Lipoproteins/immunology , Lipoproteins/physiology , Streptococcus pneumoniae/immunology , Streptococcus pneumoniae/pathogenicity , Virulence Factors/immunology , Virulence Factors/physiology , Adhesins, Bacterial/genetics , Animals , Bacterial Adhesion , Humans , Lipoproteins/genetics , Membrane Transport Proteins/genetics , Membrane Transport Proteins/immunology , Membrane Transport Proteins/physiology , Mice , Streptococcal Vaccines/immunology , Streptococcus pneumoniae/genetics , Virulence Factors/geneticsABSTRACT
Streptococcus pneumoniae (Pnc) binds to nasopharyngeal (NP) epithelial cells in the first steps of nasopharyngeal carriage and colonization through bacterial adhesins. The pneumococcal surface adhesin A (PsaA) has previously been reported to play a significant role in pneumococcal adherence and colonization. Identification of a receptor for PsaA on human epithelium will aid in understanding the pathogenesis of this bacterium. Using recombinant PsaA covalently bound to fluorescent spheres (fluospheres), we show PsaA binds to NP cells through interaction with the human cellular receptor, E-cadherin. SDS-PAGE silver stain analysis demonstrates binding of PsaA to E-cadherin. Recombinant human E-cadherin binds to and blocks PsaA-coated fluospheres and whole transparent bacteria from adhering to NP cells, but does not block a Pnc PsaA(-) mutant. Recombinant E-selectin and human alpha(5)beta(1) integrin did not bind to or block PsaA-coated fluosphere adherence to NP cells. Likewise, if NP cells were preincubated with anti-E-cadherin antibody, there was a significant decrease (46%, P=0.05) in PsaA-coated fluosphere adherence to the cells. Additionally, when using E-cadherin transfected cells, we observed PsaA-coated fluospheres bind more efficiently to cells which express E-cadherin. This work identifies E-cadherin as a receptor on human epithelial cells for the pneumococcal surface adhesin, PsaA.
Subject(s)
Adhesins, Bacterial/metabolism , Cadherins/metabolism , Lipoproteins/metabolism , Streptococcus pneumoniae/metabolism , Adhesins, Bacterial/biosynthesis , Bacterial Adhesion/physiology , Cadherins/biosynthesis , Cadherins/genetics , Calcium/metabolism , Cell Line, Tumor , Edetic Acid/pharmacology , Epithelial Cells/metabolism , Epithelial Cells/microbiology , Epithelial Cells/physiology , Humans , Lipoproteins/antagonists & inhibitors , Lipoproteins/biosynthesis , Nasopharynx/metabolism , Nasopharynx/microbiology , Nasopharynx/physiology , Pneumococcal Infections/metabolism , Pneumococcal Infections/microbiology , Streptococcus pneumoniae/drug effects , Streptococcus pneumoniae/pathogenicity , TransfectionABSTRACT
Borrelia burgdorferi undergoes differential gene expression during transmission from its tick vector to a vertebrate host. The addition of blood to a spirochete culture at 35 degrees C for 48 h had a dramatic effect on gene expression of this organism. Utilizing B. burgdorferi whole genome DNA arrays, we compared the transcriptomes of the spirochetes following a 2-day temperature shift with blood and without blood. Using combined data from three independent RNA isolations we demonstrated that the addition of blood led to a differential expression of 154 genes. Of these, 75 genes were upregulated, with 49 (65%) of them encoded on plasmids. Blood supplementation of cultures also resulted in the downregulation of 79 genes, where 56 (70%) were plasmid encoded. We verified our results by reverse transcriptase PCR of several genes in both flat and feeding ticks. In the 2-day experiment we observed the effect that exposure to increased temperature and blood combined had on B. burgdorferi gene expression at this crucial time when the spirochetes begin to move from the vector to a new vertebrate host. These changes, among others, coincide with the upregulation of the chemotaxis and sensing regulons, of the lp38-encoded ABC transporter, of proteases capable of remodeling the outer surface of the spirochetes, and of the recombination genes of cp32 as a transient or initial part of the stress response of the phage. These are all functions that could cause or facilitate the changes that spirochetes undergo following a blood meal in the tick.
Subject(s)
Bacterial Proteins/metabolism , Blood , Borrelia burgdorferi/growth & development , Gene Expression Regulation, Bacterial , Oligonucleotide Array Sequence Analysis/methods , Temperature , Animals , Bacterial Proteins/genetics , Borrelia burgdorferi/genetics , Borrelia burgdorferi/metabolism , Culture Media , Feeding Behavior , Gene Expression Profiling , Humans , Ixodes/microbiology , Ixodes/physiology , Mice , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Identification and characterization of genes that contribute to infection with Borrelia burgdorferi and, of those, genes that are targets of host responses is important for understanding the pathogenesis of Lyme disease. The complement-independent bactericidal monoclonal antibody (MAb) CB2 recognizes a carboxy-terminal, hydrophilic epitope of the outer surface protein B (OspB). CB2 kills B. burgdorferi by an unknown bactericidal mechanism. Upon binding of CB2 to OspB, differentially expressed gene products may be responsible for, or associated with, the death of the organism. A time course of the response of B. burgdorferi to CB2 was completed to analyze the differential gene expression in the bacteria over a period of visual morphological changes. Bacteria were treated with a sublethal concentration in which spirochetes were visibly distressed by the antibody but not lysed. Preliminary whole-genome DNA arrays at various time points within 1 h of incubation of B. burgdorferi with the antibody showed that most significant changes occurred at 25 min. Circular plasmid 32 (cp32)-encoded genes were active in this period of time, including the blyA homologs, phage holin system genes. DNA array data show that three blyA homologs were upregulated significantly, >/==" BORDER="0">2 standard deviations from the mean of the log ratios, and a P value of
