Immunity induced by Staphylococcus aureus surface protein A was protective against lethal challenge of Staphylococcus aureus in BALB/c mice

ABSTRACT Staphylococcus aureus is an important human pathogen that is the principal cause of a variety of diseases, ranging from localized skin infections to life-threatening systemic infections. The success of the organism as a pathogen and its ability to cause such a wide range of infections are due to its extensive virulence factors. In this study, we identified the role of the only GGDEF domain protein (GdpS [GGDEF domain protein from Staphylococcus]) in the virulence of S. aureus NCTC8325. Inactivation of gdpS results in an alteration in the production of a range of virulence factors, such as serine and cysteine proteases, fibrinogen-binding proteins, and, specifically, protein A (Spa), a major surface protein of S. aureus. The transcript level of spa decreases eightfold in the gdpS mutant compared with the parental NCTC8325 strain. Furthermore, the transcript level of sarS, which encodes a direct positive regulator of spa, also decreases in the gdpS mutant compared with the wild type, while the transcript levels of agr, sarA, sarT, and rot display no apparent changes in the gdpS mutant, suggesting that GdpS affects the expression of spa through interaction with SarS by unknown mechanisms. Furthermore, the complementation assays show that the influences of GdpS on spa and sarS depend on its N-terminal domain, which is predicted to be the sensor of a two-component system, rather than its C-terminal GGDEF domain with conserved GGDEF, suggesting that GdpS functions in S. aureus by an unknown mechanism independent of 3′,5′-cyclic diguanylic acid signaling.

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Both Innate Immunity and Type 1 Humoral Immunity to Streptococcus pneumoniae Are Mediated by MyD88 but Differ in Their Relative Levels of Dependence on Toll-Like Receptor 2

Infection and Immunity ◽

10.1128/iai.73.1.298-307.2005 ◽

2005 ◽

Vol 73 (1) ◽

pp. 298-307 ◽

Cited By ~ 74

Author(s):

Abdul Q. Khan ◽

Quanyi Chen ◽

Zheng-Qi Wu ◽

James C. Paton ◽

Clifford M. Snapper

Keyword(s):

Streptococcus Pneumoniae ◽

Humoral Immunity ◽

Protein A ◽

Surface Protein ◽

Lethal Challenge ◽

Humoral Immune ◽

Extracellular Bacterium

ABSTRACT Little is known regarding the role of Toll-like receptors (TLRs) in regulating protein- and polysaccharide-specific immunoglobulin (Ig) isotype production in response to an in vivo challenge with an extracellular bacterium. In this report we demonstrate that MyD88−/−, but not TLR2−/−, mice are markedly defective in their induction of multiple splenic proinflammatory cytokine- and chemokine-specific mRNAs after intraperitoneal (i.p.) challenge with heat-killed Streptococcus pneumoniae capsular type 14 (S. pneumoniae type 14). This is correlated with analogous responses in splenic cytokine protein release in vitro following addition of S. pneumoniae type 14. Consistent with these data, naïve MyD88−/−, but not TLR2−/−, mice are more sensitive to killing following i.p. challenge with live S. pneumoniae type 14, relative to responses in wild-type mice. However, prior immunization of MyD88−/− mice with heat-killed S. pneumoniae type 14 protects against an otherwise-lethal challenge with live S. pneumoniae type 14. Surprisingly, both MyD88−/− and TLR2−/− mice exhibit striking and equivalent defects in elicitation of type 1 IgG isotypes (IgG3, IgG2b, and IgG2a), but not the type 2 IgG isotype, IgG1, specific for several protein and polysaccharide antigens, in response to i.p. challenge with heat-killed S. pneumoniae type 14. Of note, the type 1 IgG isotype titers specific for pneumococcal surface protein A are reduced in MyD88−/− mice but not TLR2−/− mice. These data suggest that distinct TLRs may differentially regulate innate versus adaptive humoral immunity to intact S. pneumoniae and are the first to implicate a role for TLR2 in shaping an in vivo type 1 IgG humoral immune response to a gram-positive extracellular bacterium.

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Evaluation of a Vaccine Formulation against Streptococcus pneumoniae Based on Choline-Binding Proteins

Clinical and Vaccine Immunology ◽

10.1128/cvi.00692-14 ◽

2014 ◽

Vol 22 (2) ◽

pp. 213-220 ◽

Cited By ~ 5

Author(s):

Eliane N. Miyaji ◽

Cintia F. M. Vadesilho ◽

Maria Leonor S. Oliveira ◽

André Zelanis ◽

David E. Briles ◽

...

Keyword(s):

Streptococcus Pneumoniae ◽

Binding Proteins ◽

Choline Chloride ◽

Protein A ◽

Surface Protein ◽

Mass Spectrometry Analysis ◽

Effective Vaccine ◽

Lethal Challenge ◽

Content Type ◽

Spectrometry Analysis

ABSTRACTStreptococcus pneumoniaehas proteins that are attached to its surface by binding to phosphorylcholine of teichoic and lipoteichoic acids. These proteins are known as choline-binding proteins (CBPs). CBPs are an interesting alternative for the development of a cost-effective vaccine, and PspA (pneumococcal surface protein A) is believed to be the most important protective component among the different CBPs. We sought to use CBPs eluted from pneumococci as an experimental vaccine. Since PspA shows variability between isolates, we constructed strains producing different PspAs. We used the nonencapsulated Rx1 strain, which produces PspA from clade 2 (PspA2), to generate apspA-knockout strain (Rx1 ΔpspA) and strains expressing PspA from clade 1 (Rx1pspA1) and clade 4 (Rx1pspA4). We grew Rx1, Rx1 ΔpspA, Rx1pspA1, and Rx1pspA4in Todd-Hewitt medium containing 0.5% yeast extract and washed cells in 2% choline chloride (CC). SDS-PAGE analysis of the proteins recovered by a CC wash showed few bands, and the CBPs PspA and PspC (pneumococcal surface protein C) were identified by mass spectrometry analysis. Subcutaneous immunization of mice with these full-length native proteins without adjuvant led to significantly higher rates of survival than immunization with diluent after an intranasal lethal challenge with two pneumococcal strains and also after a colonization challenge with one strain. Importantly, immunization with recombinant PspA4 (rPspA4) without adjuvant did not elicit significant protection.

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Mapping of Epitopes Recognized by Antibodies Induced by Immunization of Mice with PspA and PspC

Clinical and Vaccine Immunology ◽

10.1128/cvi.00239-14 ◽

2014 ◽

Vol 21 (7) ◽

pp. 940-948 ◽

Cited By ~ 13

Author(s):

Cintia F. M. Vadesilho ◽

Daniela M. Ferreira ◽

Stephen B. Gordon ◽

David E. Briles ◽

Adriana T. Moreno ◽

...

Keyword(s):

Amino Acids ◽

Protein A ◽

Immunoglobulin A ◽

Surface Protein ◽

Factor H ◽

Peptide Arrays ◽

Pneumococcal Infections ◽

Lethal Challenge ◽

Conformational Epitopes ◽

Linear Epitopes

ABSTRACTPneumococcal surface protein A (PspA) and pneumococcal surface protein C (PspC) are important candidates for an alternative vaccine against pneumococcal infections. Since these antigens show variability, the use of variants that do not afford broad protection may lead to the selection of vaccine escape bacteria. Epitopes capable of inducing antibodies with broad cross-reactivities should thus be the preferred antigens. In this work, experiments using peptide arrays show that most linear epitopes recognized by antibodies induced in mice against different PspAs were located at the initial 44 amino acids of the mature protein and that antibodies against these linear epitopes did not confer protection against a lethal challenge. Conversely, linear epitopes recognized by antibodies to PspC included the consensus sequences involved in the interaction with human factor H and secretory immunoglobulin A (sIgA). Since linear epitopes of PspA were not protective, larger overlapping fragments containing 100 amino acids of PspA of strain Rx1 were constructed (fragments 1 to 7, numbered from the N terminus) to permit the mapping of antibodies with conformational epitopes not represented in the peptide arrays. Antibodies from mice immunized with fragments 1, 2, 4, and 5 were capable of binding onto the surface of pneumococci and mediating protection against a lethal challenge. The fact that immunization of mice with 100-amino-acid fragments located at the more conserved N-terminal region of PspA (fragments 1 and 2) induced protection against a pneumococcal challenge indicates that the induction of antibodies against conformational epitopes present at this region may be important in strategies for inducing broad protection against pneumococci.

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Expression, immunogenicity and variation of iron-regulated surface protein A from bovine isolates of Staphylococcus aureus

FEMS Microbiology Letters ◽

10.1093/femsle/fnx082 ◽

2017 ◽

Vol 364 (9) ◽

Cited By ~ 4

Author(s):

Neha Misra ◽

Tyler F. Wines ◽

Colton L. Knopp ◽

Mark A. McGuire ◽

Juliette K. Tinker

Keyword(s):

Staphylococcus Aureus ◽

Protein A ◽

Surface Protein

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MRSA screening and spa gene detection in isolates from healthcare workers at ophthalmology hospital in Egypt

Bulletin of the National Research Centre ◽

10.1186/s42269-019-0253-0 ◽

2019 ◽

Vol 43 (1) ◽

Author(s):

Maha G. Haggag ◽

Amal E. Aboelnour ◽

Mai Al-Kaffas

Keyword(s):

Staphylococcus Aureus ◽

Healthcare Workers ◽

Protein A ◽

Healthcare Providers ◽

Surface Protein ◽

Gene Detection ◽

Eye Infections ◽

Mrsa Strains ◽

Spa Gene ◽

Research Institute

Abstract Background Staphylococcus aureus has a major role in different types of eye infections as conjunctivitis, keratitis, and endophthalmitis. Methicillin-resistant Staphylococcus aureus (MRSA) was almost restricted to hospitals, but its prevalence has been increased in people outside hospitals. The cell wall of Staphylococcus aureus has protein A which can bind to the Fc portion of IgG. This ptnA is encoded by surface protein A of Staphylococcus aureus (spa) gene that contains a highly polymorphic sequence which is composed of repeats of 24-bp. Sequence typing of the spa gene repeat region is used to study the epidemiology of MRSA. The purpose of this study was screening of MRSA strains among healthcare workers (HCWs) in the Hospital of the Research Institute of Ophthalmology (RIO), Giza, Egypt, and detecting spa gene in their DNAs by PCR. Results In the present study, 81 samples from healthcare providers in the hospital of the Research Institute of Ophthalmology, Egypt, were screened for MRSA. Out of these 81 samples, 41 isolates (50.6%) were identified as coagulase-positive Staphylococcus aureus. Twelve staphylococcal isolates were resistant to both oxacillin and cefoxitin, and those were identified as MRSA with a percentage of 14.8% (12/81). Conventional PCR could detect spa gene in 10 out of 12 DNA MRSA with a percentage of 83.3% (10/12). Conclusion In the present study, the prevalence of MRSA in HCWs was 14.8%. Since amplification of spa gene by PCR is a necessary preliminary step for spa typing of MRSA and since using different primers for spa gene amplification might affect PCR results, then proper selection of the primers and thermal cycling reaction conditions are recommended for PCR performance and spa typing.

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Iron-Regulated Surface Determinant Protein A Mediates Adhesion of Staphylococcus aureus to Human Corneocyte Envelope Proteins

Infection and Immunity ◽

10.1128/iai.01304-08 ◽

2009 ◽

Vol 77 (6) ◽

pp. 2408-2416 ◽

Cited By ~ 56

Author(s):

Simon R. Clarke ◽

Guillaume Andre ◽

Evelyn J. Walsh ◽

Yves F. Dufrêne ◽

Timothy J. Foster ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Single Molecule ◽

Protein A ◽

Surface Protein ◽

High Specificity ◽

Human Infection ◽

Cell Binding ◽

Single Molecule Force Spectroscopy ◽

Whole Cell ◽

Cornified Envelope

ABSTRACT The ability of Staphylococcus aureus to colonize the human nares is a crucial prerequisite for disease. IsdA is a major S. aureus surface protein that is expressed during human infection and required for nasal colonization and survival on human skin. In this work, we show that IsdA binds to involucrin, loricrin, and cytokeratin K10, proteins that are present in the cornified envelope of human desquamated epithelial cells. To measure the forces and dynamics of the interaction between IsdA and loricrin (the most abundant protein of the cornified envelope), single-molecule force spectroscopy was used, demonstrating high-specificity binding. IsdA acts as a cellular adhesin to the human ligands, promoting whole-cell binding to immobilized proteins, even in the absence of other S. aureus components (as shown by heterologous expression in Lactococcus lactis). Inhibition experiments revealed the binding of the human ligands to the same IsdA region. This region was mapped to the NEAT domain of IsdA. The NEAT domain also was found to be required for S. aureus whole-cell binding to the ligands as well as to human nasal cells. Thus, IsdA is an important adhesin to human ligands, which predominate in its primary ecological niche.

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A protein component of the cell surface of Staphylococcus aureus

Biochemical Journal ◽

10.1042/bj1080257 ◽

1968 ◽

Vol 108 (2) ◽

pp. 257-262 ◽

Cited By ~ 8

Author(s):

A. M. James ◽

J. E. Brewer

Keyword(s):

Staphylococcus Aureus ◽

Protein A ◽

Surface Protein ◽

Protein Component ◽

The Other ◽

Animal Origin ◽

Aureus Strain ◽

Similar Treatment ◽

Ph Values ◽

Protein Components

1. Treatment with trypsin of cells of Staphylococcus aureus strain Cowan I, known to carry a protein component that includes the Jensen protein A, results in an increase in the negative mobility of the cells at pH values greater than 6. 2. Similar treatment of cells of strain Wood 46, which has no surface protein component, causes no change in the electrophoretic mobility. 3. Electrokinetically heterogeneous populations are observed in two strains of S. aureus, one of human and the other of animal origin. Evidence is presented ascribing this to the presence of varying amounts of protein components on the surface of different cells.

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