lipoteichoic-acid and Endocarditis--Bacterial

Enterococcus faecium (E. faecium) has emerged as one of today's leading causes of health care-associated infections that is difficult to treat with the available antibiotics. These pathogens produce capsular polysaccharides on the cell surface which play a significant role in adhesion, virulence and evasion. Therefore, we aimed at the identification and characterization of bacterial polysaccharide antigens which are central for the development of vaccine-based prophylactic approaches. The crude cell wall-associated polysaccharides from E. faecium, its mutant and complemented strains were purified and analyzed by a primary antibody raised against lipoteichoic acid (LTA) and diheteroglycan (DHG). The resistant E. faecium strains presumably possess novel capsular polysaccharides that allow them to avoid the evasion from opsonic killing. The E. faecium U0317 strain was very well opsonized by anti-U0317 (~95%), an antibody against the whole bacterial cell. The deletion mutant showed a significantly increased susceptibility to opsonophagocytic killing (90-95%) against the penicillin binding protein (anti-PBP-5). By comparison, in a mouse urinary tract and rat endocarditis infection model, respectively, there were no significant differences in virulence. In this study we explored the biological role of the capsule of E. faecium. Our findings showed that the U0317 strain is not only sensitive to anti-LTA but also to antibodies against other enterococcal surface proteins. Our findings demonstrate that polysaccharides capsule mediated-resistance to opsonophagocytosis. We also found that the capsular polysaccharides do not play an important role in bacterial virulence in urinary tract and infective endocarditis in vivo models.

Topics: Animals; Anti-Bacterial Agents; Antibodies, Bacterial; Antigens, Bacterial; Bacterial Capsules; Cell Wall; Disease Models, Animal; Drug Resistance, Bacterial; Endocarditis, Bacterial; Enterococcus faecium; Female; Gram-Positive Bacterial Infections; Humans; Leukocytes, Mononuclear; Lipopolysaccharides; Mice; Mice, Inbred BALB C; Opsonin Proteins; Penicillin-Binding Proteins; Phagocytosis; Polysaccharides, Bacterial; Primary Cell Culture; Rats; Rats, Wistar; Teichoic Acids; Urinary Tract Infections

Infective endocarditis is usually caused by Streptococcus sanguinis and characterized by inflammatory responses in the endocardium. This study aimed to investigate if the new compound (3R)-5,6,7-trihydroxy-3-isopropyl-3-methylisochroman-1-one (TIM) isolated from Alpinia katsumadai Hayata could provide protection against lipoteichoic acid (LTA)-induced cell damage in embryonic rat heart cells (H9c2).. LTA-induced cell damage was established in H9c2, and the protective effects of TIM against the cell damage were examined at different concentrations (0.1-2.5 µM). The inflammatory response and oxidative stress in H9c2 cells were also measured.. Treatment with TIM (0.1-2.5 µM) significantly decreased LTA-induced toxicity in H9c2 cells, which was indicated by increase in cell viability, elevation in the mitochondrial membrane potential, decrease in the release of cytochrome-c and DNA damage, inhibition of caspase-3/9 activities, and change in apoptosis-related protein expression in LTA-treated H9c2 cells. TIM treatment also significantly attenuated the redox imbalance in H9c2 cells by decreasing malondialdehyde and intracellular reactive oxygen species levels as well as by enhancing superoxide dismutase activities and glutathione levels by increasing nuclear factor (erythroid-derived 2)-like 2 protein expression. Moreover, TIM treatment decreased interleukin 1 ß, interleukin 12, and tumor necrosis factor α levels by inhibiting nuclear factor kappa B protein expression.. Our data indicated that TIM protected H9c2 cells against LTA-induced toxicity, at least partially through inhibiting the inflammatory response and oxidative stress, providing scientific rational to develop TIM to treat infective endocarditis.

Topics: Animals; Antioxidants; Cardiotoxicity; Cell Line; Chromans; Disease Models, Animal; Endocarditis, Bacterial; Lipopolysaccharides; Myoblasts, Cardiac; Rats; Streptococcus; Teichoic Acids

Infective endocarditis is caused by Streptococcus sanguinis present in dental plaque, which can induce inflammatory responses in the endocardium. The present study depicts research on the properties of apigenin in embryonic mouse heart cells (H9c2) treated with lipoteichoic acid (LTA) obtained from S. sanguinis. Interleukin-1β and cyclooxygenase (COX)-2 expression were detected by reverse transcriptase polymerase chain reaction. In addition, western blot assays and immuno-fluorescence staining were used to assess translocation of nuclear factor kappa beta (NF-κB), degradation of IκB, as well as activity of the mitogen activated protein kinases: extracellular signal-regulated kinase (ERK)1/2, p38, and c-Jun N-terminal kinase (JNK). Effect of apigenin on cell viability was equally assessed in other experimental series. Our results showed that apigenin blocked activation of ERK, JNK, and p38 in cardiomyocytes treated with LTA in a dose-dependent fashion. Moreover, apigenin showed no cytotoxic effects; it blocked NF-κB translocation and IκB degradation. Our findings suggested that apigenin possessed potential value in the treatment of infectious endocarditis.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Apigenin; Cell Survival; Cells, Cultured; Cyclooxygenase 2; Endocarditis, Bacterial; I-kappa B Kinase; Interleukin-1beta; JNK Mitogen-Activated Protein Kinases; Lipopolysaccharides; Mice; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Myoblasts, Cardiac; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Rats; Signal Transduction; Streptococcus sanguis; Teichoic Acids

Infective endocarditis is caused by bacterial colonization of the endocardium. Because endocardium modulates mechanical performance of subjacent myocardium, we studied acute effects of bacteria on isolated cardiac muscle and on the functional role of the endocardium. Bacteria, grown in broth at 37 degrees C, were added at increasing concentrations (10(2) to 10(6) bacteria/ml) to cat papillary muscles in Krebs-Ringer solution (1.25 mM Ca2+, 35 degrees C). The endocardial surface was damaged by exposing muscles to a stream of dry air for 30 s. Streptococcus (Enterococcus) faecalis induced significant increases in total peak isometric twitch tension (TT) and maximal velocity of unloaded shortening (Vmax) and significant decreases in time to TT (TtTT) and time to half isometric twitch tension decline (RT 1/2), both before and after removal of endocardial endothelium. This response could also be elicited with bacterial filtrate, after boiling the filtrate or after extracting the polysaccharides from it with KIO4. Increasing Ca2+ concentrations progressively reduced the response to the filtrate. Propranolol slightly, although not significantly, diminished the effects on TT and Vmax while abolishing the effects on TtTT and on RT 1/2. By contrast, Streptococcus bovis and Staphylococcus aureus did not affect TT or Vmax but induced a slight but significant decrease in TtTT at the highest concentration of bacteria. Accordingly, the filtrate of Strep. faecalis induces a positive inotropic effect. The active component is neither a protein nor a polysaccharide, and its effect may be partly beta-adrenoceptor mediated. Strep. bovis and Staph. aureus have negligible acute effects on contractility.

Topics: Adrenergic beta-Antagonists; Animals; Arrhythmias, Cardiac; Calcium; Cats; Endocarditis, Bacterial; Endocardium; Endothelium; Enterococcus faecalis; Hot Temperature; Kinetics; Lipopolysaccharides; Microscopy, Electron, Scanning; Myocardial Contraction; Polysaccharides; Propranolol; Stimulation, Chemical; Streptococcal Infections; Teichoic Acids

Invasion by gram-positive and gram-negative bacterial organisms is characterized immunopathologically by the activation of mononuclear phagocytic cells, leading to the elaboration of macrophage-derived regulatory and chemotactic factors, and the resultant influx of inflammatory leukocytes. Little is known regarding the mechanisms by which gram-positive organisms initiate macrophage activation and subsequent inflammation. In this investigation, we postulated that lipoteichoic acid (LTA) purified from two different gram-positive bacterial species was an important signal for the expression of chemotactic cytokines from human peripheral blood monocytes (PBM). In initial experiments, we demonstrated that cell-associated interleukin-8 (IL-8) was expressed by mononuclear phagocytes present in inflamed areas of endocardium in cases of acute Staphylococcus aureus endocarditis. We next demonstrated that LTA purified from either Staphylococcus aureus or Streptococcus pyogenes induced the time- and dose-dependent expression of IL-8 mRNA and protein from human PBM. The expression of IL-8 mRNA from LTA- but not lipopolysaccharide (LPS)-treated PBM was superinduced by concomitant treatment with cycloheximide, indicating that the expression of IL-8 mRNA from LTA-treated PBM was negatively controlled by repressor proteins. Furthermore, mRNA stability studies indicated that IL-8 mRNA was less stable in the presence of LTA than in the presence of LPS. Our findings indicate that LTA can induce the secretion of the polymorphonuclear leukocyte chemotactic factor IL-8 and that LTA may be an important cellular mediator of inflammatory cell recruitment that characterizes immune responses to gram-positive bacterial infections.

Topics: Cycloheximide; Endocarditis, Bacterial; Gene Expression; Humans; In Vitro Techniques; Interleukin-8; Lipopolysaccharides; Monocytes; RNA, Messenger; Staphylococcal Infections; Staphylococcus aureus; Streptococcus pyogenes; Teichoic Acids

An enzyme-linked immunoassay (ELISA) was used to detect antibodies in human sera to Staphylococcus aureus peptidoglycan (PG) and lipoteichoic acid (LTA). All the sera from the blood donors contained IgG antibodies to both substances. Among the sera from 34 patients with bacteriologically verified, serious S. aureus infections, 71 per cent contained significantly elevated levels of anti-PG antibodies and 76 per cent of anti-LTA antibodies. Among the sera from 38 patients with suspected but not bacteriologically verified staphylococcal infections, 58 per cent contained significantly elevated levels of anti-PG antibodies and 74 per cent of anti-LTA antibodies. The levels of antibodies to PG correlated well with the levels of antibodies to LTA, but the latter occurred over a broader range in the patient sera. Elevated antibody values were, however, also found in some patients with serious, non-staphylococcal infections. The diagnostic value of PG and LTA antibodies has to be further investigated.

Topics: Antibodies, Bacterial; Blood Donors; Endocarditis, Bacterial; Enzyme-Linked Immunosorbent Assay; Humans; Immunoglobulin G; Lipopolysaccharides; Peptidoglycan; Phosphatidic Acids; Staphylococcal Infections; Staphylococcus aureus; Teichoic Acids

Sera from individuals with Staphylococcus aureus endocarditis and osteomyelitis and from some individuals with other forms of gram-positive endocarditis yielded higher readings in a microenzyme-linked immunosorbent assay against lipoteichoic acid from S. aureus than did sera from individuals with other types of serious staphylococcal infection or non-staphylococcal osteomyelitis, or from unselected inpatients.

Topics: Antibodies, Bacterial; Antigens, Bacterial; Endocarditis, Bacterial; Enzyme-Linked Immunosorbent Assay; Humans; Lactobacillus; Lipopolysaccharides; Osteomyelitis; Phosphatidic Acids; Proteus Infections; Pseudomonas Infections; Staphylococcal Infections; Staphylococcus aureus; Streptococcal Infections; Teichoic Acids

Topics: Antibodies, Bacterial; Endocarditis, Bacterial; Humans; Lipopolysaccharides; Osteomyelitis; Peptidoglycan; Phosphatidic Acids; Sepsis; Staphylococcal Infections; Staphylococcus aureus; Teichoic Acids

The effect of penicillin treatment of Streptococcus sanguis in vitro, on subsequent bacterial density in the bloodstream and on cardiac valves in the rabbit model of endocarditis was studied. As experimental tools for this study, isogenic pairs of S. sanguis differing in resistance to streptomycin or rifampin were prepared by genetic transformation. Rabbits with traumatized heart valves received an intravenous inoculation of penicillin treated (1 mug/ml) and untreated S. sanguis, each marked by resistance to either streptomycin or rifampin. The number of penicillin-treated and untreated bacteria attached to the valvular surfaces was determined by differential counting on streptomycin or rifampin containing media. Penicillin pretreatment reduced cardiac valve colonization 5 min after inoculation ("adherence ratio" x 10(8) was 4.11 for the control and 3.66 for the penicillin-treated bacteria, P < 0.001). The results were not due to differences in serum killing or bacterial densities in the bloodstream. There was no difference in valvular bacterial densities 24 h after bacterial inoculation (adherence ratio x 10(8), 7.26 untreated vs. 6.34 penicillin-pretreated, P > 0.10). In vitro experiments were performed using platelet-fibrin surfaces to test the possibility that penicillin-induced loss of lipoteichoic acid was responsible for decreased streptococcal adherence. Pretreatment of S. sanguis cultures with inhibitory concentrations of penicillin or with antiserum against lipoteichoic acid and precoating of the platelet-fibrin surfaces with lipoteichoic acid, all caused reduction in bacterial adherence. The findings are interpreted as support for the role of lipoteichoic acid as an adhesin in S. sanguis interactions with particular host tissue surfaces.

Topics: Adhesiveness; Animals; Disease Models, Animal; Endocarditis, Bacterial; Heart Valves; Lipopolysaccharides; Microbial Sensitivity Tests; Penicillins; Phosphatidic Acids; Rabbits; Streptococcal Infections; Streptococcus sanguis; Teichoic Acids