scholarly journals Generation and Properties of a Streptococcus pneumoniae Mutant Which Does Not Require Choline or Analogs for Growth

1998 ◽  
Vol 180 (8) ◽  
pp. 2093-2101 ◽  
Author(s):  
Janet Yother ◽  
Klaus Leopold ◽  
Johanna White ◽  
Werner Fischer
Keyword(s):  
Streptococcus Pneumoniae ◽  
Stationary Phase ◽  
Cell Walls ◽  
Parent Strain ◽  
Protein A ◽  
Teichoic Acid ◽  
Surface Protein ◽  
Chain Structure ◽  
Wall Teichoic Acid ◽  
Mutant Cells

A mutant (JY2190) of Streptococcus pneumoniae Rx1 which had acquired the ability to grow in the absence of choline and analogs was isolated. Lipoteichoic acid (LTA) and wall teichoic acid (TA) isolated from the mutant were free of phosphocholine and other phosphorylated amino alcohols. Both polymers showed an unaltered chain structure and, in the case of LTA, an unchanged glycolipid anchor. The cell wall composition was also not altered except that, due to the lack of phosphocholine, the phosphate content of cell walls was half that of the parent strain. Isolated cell walls of the mutant were resistant to hydrolysis by pneumococcal autolysin (N-acetylmuramyl-l-alanine amidase) but were cleaved by the muramidases CPL and cellosyl. The lack of active autolysin in the mutant cells became apparent by impaired cell separation at the end of cell division and by resistance against stationary-phase and penicillin-induced lysis. As a result of the absence of choline in the LTA, pneumococcal surface protein A (PspA) was no longer retained on the cytoplasmic membrane. During growth in the presence of choline, which was incorporated as phosphocholine into LTA and TA, the mutant cells separated normally, did not release PspA, and became penicillin sensitive. However, even under these conditions, they did not lyse in the stationary phase, and they showed poor reactivity with antibody to phosphocholine and an increased release of C-polysaccharide from the cell. In contrast to ethanolamine-grown parent cells (A. Tomasz, Proc. Natl. Acad. Sci. USA 59:86–93, 1968), the choline-free mutant cells retained the capability to undergo genetic transformation but, compared to Rx1, with lower frequency and at an earlier stage of growth. The properties of the mutant could be transferred to the parent strain by DNA of the mutant.

scholarly journals 4-1BB (CD137) Differentially Regulates Murine In Vivo Protein- and Polysaccharide-Specific Immunoglobulin Isotype Responses to Streptococcus pneumoniae

2003 ◽  
Vol 71 (1) ◽  
pp. 196-204 ◽  
Author(s):  
Zheng-Qi Wu ◽  
Abdul Q. Khan ◽  
Yi Shen ◽  
Karen M. Wolcott ◽  
Wojciech Dawicki ◽  
...  
Keyword(s):  
T Cells ◽  
Streptococcus Pneumoniae ◽  
Protein A ◽  
Teichoic Acid ◽  
Surface Protein ◽  
Cell Receptor ◽  
Primary Response ◽  
Humoral Immune ◽  
Specific Immunoglobulin

ABSTRACT 4-1BB (CD137) is induced on activated CD4+ and CD8+ T cells and delivers a costimulatory signal upon binding the 4-1BB ligand (4-1BBL) expressed on antigen-presenting cells. Induction of 4-1BB is dependent on activation via the T-cell receptor (TCR) and possibly CD28. It was previously demonstrated that both an in vivo protein (pneumococcal surface protein A [PspA])- and polysaccharide (phosphorylcholine [PC] determinant of teichoic acid)-specific immunoglobulin (Ig) isotype response to Streptococcus pneumoniae was dependent on CD4+ TCRαβ+ T cells and B7-dependent costimulation through CD28. We thus postulated that 4-1BB costimulation would also play a role in regulating the in vivo anti-PspA and anti-PC response to S. pneumoniae. We demonstrate that mice genetically deficient in 4-1BBL elicit a markedly reduced IgM and IgG anti-PC but normal primary and secondary IgG anti-PspA responses to S. pneumoniae relative to those for wild-type mice. However, injection of an agonistic anti-4-1BB monoclonal antibody (MAb), while having no significant effect on the anti-PC response, strongly inhibits the primary anti-PspA response, the generation of PspA-specific memory, and germinal center formation but does not induce a lasting state of tolerance. In contrast, anti-4-1BB MAb has no effect on the anti-PspA response when injected only at the time of secondary immunization. Delay of the addition of anti-4-1BB leads to progressively less inhibition of the primary response up to day 8. This inhibition is independent of CD8+ T cells and is associated with the expansion of CD4+ T cells with an activated phenotype, which is partly dependent on B7-dependent costimulation. These data are the first to suggest a stimulatory role for endogenous 4-1BB-4-1BBL interactions during a humoral immune response to a pathogen and further underscore significant differences in costimulation requirements for an in vivo protein- versus polysaccharide-specific Ig isotype response to an extracellular bacterium.

The physiology of teichoic acid deficient staphylococci

1973 ◽  
Vol 19 (11) ◽  
pp. 1393-1399 ◽  
Author(s):  
Li-Tse Ou ◽  
A. N. Chatterjee ◽  
F. E. Young ◽  
R. E. Marquis
Keyword(s):  
Cell Walls ◽  
Parent Strain ◽  
Teichoic Acid ◽  
Low Ph ◽  
Aureus Strain ◽  
Binding Affinities ◽  
Inhibitory Effects ◽  
Growth Inhibitory ◽  
Acid Titration ◽  
Phosphate Groups

Cell walls isolated from a teichoic acid deficient mutant (52A5) of Staphylococcus aureus strain H were found to have lower capacities to bind cations than did walls of the parent strain. Both types of walls had higher binding affinities for Mg2+ and Ca2+ than for K+ and Na+. The reduced number of phosphate groups in 52A5 walls was reflected in a higher apparent pKa of 4.3 for displacement of Mg2+ (or Ca2+) during acid titration with HCl. The comparable pKa value for displacement of bound Mg2+ from parent-strain walls was 3.7. The reduced capacity of 52A5 walls to bind cations was not reflected in any significant increase in sensitivity to the growth inhibitory actions of ethylenediaminetetraacetate, low pH, or high NaCl concentrations. However, the 52A5 strain was somewhat more sensitive to the inhibitory effects of high pH. Also, mutant walls were found to be structurally more compact than walls of the parent strain, presumably because of less extensive electrostatic repulsion within the wall matrix.

scholarly journals DNA vaccines expressing pneumococcal surface protein A (PspA) elicit protection levels comparable to recombinant protein

2006 ◽  
Vol 55 (4) ◽  
pp. 375-378 ◽  
Author(s):  
Daniela M. Ferreira ◽  
Eliane N. Miyaji ◽  
Maria Leonor S. Oliveira ◽  
Michelle Darrieux ◽  
Ana Paula M. Arêas ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Recombinant Protein ◽  
Dna Vaccines ◽  
Protein A ◽  
Surface Protein ◽  
Humoral Response ◽  
Cost Effective ◽  
Promising Candidate ◽  
Pneumococcal Surface Protein A ◽  
Antibody Levels

Pneumococcal surface protein A (PspA) is a promising candidate for the development of cost-effective vaccines against Streptococcus pneumoniae. In the present study, BALB/c mice were immunized with DNA vaccine vectors expressing the N-terminal region of PspA. Animals immunized with a vector expressing secreted PspA developed higher levels of antibody than mice immunized with the vector expressing the antigen in the cytosol. However, both immunogens elicited similar levels of protection against intraperitoneal challenge. Furthermore, immunization with exactly the same fragment in the form of a recombinant protein, with aluminium hydroxide as an adjuvant, elicited even higher antibody levels, but this increased humoral response did not correlate with enhanced protection. These results show that DNA vaccines expressing PspA are able to elicit protection levels comparable to recombinant protein, even though total anti-PspA IgG response is considerably lower.

scholarly journals Interconnection of competence, stress and CiaR regulons in Streptococcus pneumoniae: competence triggers stationary phase autolysis of ciaR mutant cells

2004 ◽  
Vol 51 (4) ◽  
pp. 1071-1086 ◽  
Author(s):  
Adilia Dagkessamanskaia ◽  
Miriam Moscoso ◽  
Vincent Hénard ◽  
Sébastien Guiral ◽  
Karin Overweg ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Stationary Phase ◽  
Mutant Cells

scholarly journals Determination of phenotypes and pneumococcal surface protein A family types of Streptococcus pneumoniae from Malaysian healthy children

2013 ◽  
Vol 46 (3) ◽  
pp. 180-186 ◽  
Author(s):  
Masura Mohd Yatim ◽  
Siti Norbaya Masri ◽  
Mohd Nasir Mohd Desa ◽  
Niazlin Mohd Taib ◽  
Syafinaz Amin Nordin ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Protein A ◽  
Surface Protein ◽  
Healthy Children ◽  
Pneumococcal Surface Protein A ◽  
Family Types

scholarly journals Immunization of Mice with Combinations of Pneumococcal Virulence Proteins Elicits Enhanced Protection against Challenge with Streptococcus pneumoniae

2000 ◽  
Vol 68 (5) ◽  
pp. 3028-3033 ◽  
Author(s):  
A. David Ogunniyi ◽  
Rebekah L. Folland ◽  
David E. Briles ◽  
Susan K. Hollingshead ◽  
James C. Paton
Keyword(s):  
Streptococcus Pneumoniae ◽  
Metal Binding ◽  
Protein A ◽  
Surface Protein ◽  
Active Immunization ◽  
Survival Times ◽  
Virulence Proteins ◽  
Virulent Type ◽  
Vaccine Potential ◽  
High Doses

ABSTRACT The vaccine potential of a combination of three pneumococcal virulence proteins was evaluated in an active-immunization–intraperitoneal-challenge model in BALB/c mice, using very high challenge doses of Streptococcus pneumoniae. The proteins evaluated were a genetic toxoid derivative of pneumolysin (PdB), pneumococcal surface protein A (PspA), and a 37-kDa metal-binding lipoprotein referred to as PsaA. Mice immunized with individual proteins or combinations thereof were challenged with high doses of virulent type 2 or type 4 pneumococci. The median survival times for mice immunized with combinations of proteins, particularly PdB and PspA, were significantly longer than those for mice immunized with any of the antigens alone. A similar effect was seen in a passive protection model. Thus, combinations of pneumococcal proteins may provide the best non-serotype-dependent protection against S. pneumoniae.

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