Group B streptococcal septicemia and delayed-onset congenital right-sided diaphragmatic hernia.

A case is reported of fulminant early-onset group B streptococcal septicemia and delayed-onset congenital right-sided diaphragmatic hernia in a neonate. The latter condition should be considered when early-onset group B streptococcal disease is followed by increasing respiratory distress, right-sided pleural effusion and partial or complete opacification of the right side of the thorax.     

Targeting the BspC-vimentin interaction to develop anti-virulence therapies during Group B streptococcal meningitis

Bacterial infections are a major cause of morbidity and mortality worldwide and the rise of antibiotic resistance necessitates development of alternative treatments. Pathogen adhesins that bind to host cells initiate disease pathogenesis and represent potential therapeutic targets. We have shown previously that the BspC adhesin in Group B Streptococcus (GBS), the leading cause of bacterial neonatal meningitis, interacts with host vimentin to promote attachment to brain endothelium and disease development. Here we determined that the BspC variable (V-) domain contains the vimentin binding site and promotes GBS adherence to brain endothelium. Site directed mutagenesis identified a binding pocket necessary for GBS host cell interaction and development of meningitis. Using a virtual structure-based drug screen we identified compounds that targeted the V-domain binding pocket, which blocked GBS adherence and entry into the brain in vivo. These data indicate the utility of targeting the pathogen-host interface to develop anti-virulence therapeutics.     

Serum opacity factor promotes group A streptococcal epithelial cell invasion and virulence

Serum opacity factor (SOF) is a bifunctional cell surface protein expressed by 40-50% of group A streptococcal (GAS) strains comprised of a C-terminal domain that binds fibronectin and an N-terminal domain that mediates opacification of mammalian sera. The sof gene was recently discovered to be cotranscribed in a two-gene operon with a gene encoding another fibronectin-binding protein, sfbX. We compared the ability of a SOF(+) wild-type serotype M49 GAS strain and isogenic mutants lacking SOF or SfbX to invade cultured HEp-2 human pharyngeal epithelial cells. Elimination of SOF led to a significant decrease in HEp-2 intracellular invasion while loss of SfbX had minimal effect. The hypoinvasive phenotype of the SOF(-) mutant could be restored upon complementation with the sof gene on a plasmid vector, and heterologous expression of sof49 in M1 GAS or Lactococcus lactis conferred marked increases in HEp-2 cell invasion. Studies using a mutant sof49 gene lacking the fibronectin-binding domain indicated that the N-terminal opacification domain of SOF contributes to HEp-2 invasion independent of the C-terminal fibronectin binding domain, findings corroborated by observations that a purified SOF N-terminal peptide could promote latex bead adherence to HEp-2 cells and inhibit GAS invasion of HEp-2 cells in a dose-dependent manner. Finally, the first in vivo studies to employ a single gene allelic replacement mutant of SOF demonstrate that this protein contributes to GAS virulence in a murine model of necrotizing skin infection.     

Novel protein vaccine candidates against Group B streptococcal infection identified using alkaline phosphatase fusions

Using an alkaline phosphatase-based genetic screening method, we identified a number of proteins that are potentially located on the outer surface of Group B streptococcus (Streptococcus agalactiae). In an enzyme-linked immunosorbent assay, antisera raised against two of the proteins, the streptococcal yutD homologue and a subunit of an ABC transporter, recognised clinically important serotypes of Group B streptococcus. In a neonatal rat model, purified IgG from the sera conferred significant levels of protection against a lethal challenge infection. The proteins identified show potential as protein subunit candidates for vaccines against Group B streptococcal disease in neonates.     

Group B streptococcal capsular sialic acids interact with siglecs (immunoglobulin-like lectins) on human leukocytes

Group B Streptococcus (GBS) is classified into nine serotypes that vary in capsular polysaccharide (CPS) architecture but share in common the presence of a terminal sialic acid (Sia) residue. This position and linkage of GBS Sia closely resembles that of cell surface glycans found abundantly on human cells. CD33-related Siglecs (CD33rSiglecs) are a family of Sia-binding lectins expressed on host leukocytes that engage host Sia-capped glycans and send signals that dampen inflammatory gene activation. We hypothesized that GBS evolved to display CPS Sia as a form of molecular mimicry limiting the activation of an effective innate immune response. In this study, we applied a panel of immunologic and cell-based assays to demonstrate that GBS of several serotypes interacts in a Sia- and serotype-specific manner with certain human CD33rSiglecs, including hSiglec-9 and hSiglec-5 expressed on neutrophils and monocytes. Modification of GBS CPS Sia by O acetylation has recently been recognized, and we further show that the degree of O acetylation can markedly affect the interaction between GBS and hSiglec-5, -7, and -9. Thus, production of Sia-capped bacterial polysaccharide capsules that mimic human cell surface glycans in order to engage CD33rSiglecs may be an example of a previously unrecognized bacterial mechanism of leukocyte manipulation.     

Group B streptococcal pilus proteins contribute to adherence to and invasion of brain microvascular endothelial cells

Surface filamentous structures known as pili have been discovered recently in the gram-positive streptococcal pathogens that cause invasive disease in humans, including group B Streptococcus (GBS). We show that two GBS proteins involved in pilus formation, encoded by pilA and pilB, also facilitate the interaction of this important agent of central nervous system infection with endothelial cells of the human blood-brain barrier.     

Truncated variants of apolipoprotein B cause hypobetalipoproteinaemia.

Familial hypobetalipoproteinaemia is a rare autosomal dominant disorder in which levels of apo-B-containing plasma lipoproteins are approximately half-normal in heterozygotes and virtually absent in homozygotes. Here we describe mutations of the apo-B gene that cause two different truncated variants of apo-B in unrelated individuals with hypobetalipoproteinaemia. One variant, apo-B(His1795----Met-Trp-Leu-Val-Thr-Term) is predicted to be 1799 amino acids long and arises from deletion of a single nucleotide (G) from leucine codon 1794. This protein was found at low levels in very low density and low density lipoprotein fractions in the blood. The second, shorter variant, apo-B(Arg1306----Term), is caused by mutation of a CpG dinucleotide in arginine codon 1306 converting it to a stop codon and predicting a protein of 1305 residues. The product of this allele could not be detected in the circulation. The differences in size and behaviour of these two variants compared to apo-B100 or apo-B48 point to domains that may be important for the assembly, secretion or stability of apo-B-containing lipoproteins.     

Pathogenesis of neonatal group B streptococcal infections

Infections of the neonate due to the group B Streptococcus have been recognized since the 1930s, but it was during the 1970s that their incidence grew alarmingly throughout the world. A research effort stimulated by this problem has yielded significant new information about many facets of the pathogenesis of these infections. Immunologic investigations have pinpointed a lack of transplacentally acquired antibody as a significant risk factor. In the laboratory, assays of antibody which have a functional endpoint have demonstrated that the type-specific carbohydrate antigens play a major role in stimulating the development of protective antibody. These assays have been shown to correlate with certain tests of primary antigen-antibody interaction which do not have a functional endpoint, but are simpler to use in larger scale epidemiologic studies. These tools may be useful in filling the gaps in our current knowledge of the pathogenesis of this infection.     

Molecular epidemiology of group B streptococcal infections

Streptococcus agalactiae (GBS) is a causative agent of sepsis and meningitis in newborns and diseases in pregnant women and nonpregnant adults. Various approaches, including both nongenetic and genetic techniques, are currently used for the study of epidemiology of GBS infections. In the present paper the different methods of molecular epidemiology of GBS infections are reviewed, and several novel approaches are introduced. The advantages and disadvantages of molecular methods are discussed and compared with traditional serotyping technique. The possible use of the molecular approaches for identification of different genetic lineages in GBS as well as for identification and control of the epidemiologically actual clones is discussed.     

Group A beta-hemolytic streptococcal glossal necrotizing myositis--case report and review

We report the first case of glossal necrotizing myositis by group A beta-hemolytic Streptococcus in an 8-year-old girl on chronic nonsteroidal anti-inflammatory drugs, immunomodulators, and steroids for juvenile rheumatoid arthritis. Treatment included partial glossectomy and parenteral antibiotics. After a critical course, full recovery ensued. The subject of necrotizing myositis is reviewed.     

Ultrastructural changes during propagation of a Group D streptococcal L-form

Summary L-form colonies derived from a Group D streptococcus showed alterations in fine structure during propagation. For the first 6 months after preparation the L-forms were capable of reversion and fibrillar and lamellate inclusions were found in some cells. Fibrillar bundles, composed of fibrils with an external diameter of 12.5 nm were found attached to the cytoplasmic membrane of the large 20 m surface cells. Small, 1 m cells within the agar contained lamellate inclusions with a periodicity of 14 nm, also attached to the cytoplasmic membrane. In empty cells both inclusions appeared as hollow fibrils. The composition of these structures is not known. Long filamentous evaginations of the cytoplasmic membrane were also found in a small proportion of cells. After 15 months' propagation, when the L-form had become stable, the inclusions and filaments were no longer visible.