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PubMed2000

Opsonic potential, protective capacity, and sequence conservation of the Treponema pallidum subspecies pallidum Tp92.

By means of a differential screening technique, a 92-kDa antigen, designated Tp92, was identified from Treponema pallidum subspecies pallidum. This protein is similar in sequence to the protective surface antigens D15 from Haemophilus influenzae and Oma87 from Pasteurella multocida. Amino acid sequence analyses revealed a cleavable N-terminal signal sequence and predicted the outer membrane location for Tp92. In support of this, antiserum raised against recombinant Tp92 promotes opsonization and phagocytosis of T. pallidum by rabbit macrophages, and anti-Tp92 reactivity is absent from washed treponemal preparations presumed to be lacking outer membranes. The Tp92 amino acid sequence is 95.5%-100% conserved among 11 strains representing 4 pathogenic treponemes, and immunization with recombinant Tp92 partially protected rabbits from subsequent T. pallidum challenge. These results demonstrate that Tp92 is an invariant, immunoprotective antigen that may be present on the surface of T. pallidum and may represent a potential vaccine candidate for syphilis.

Amino Acid Sequence↗
PubMed2001

Molecular subtyping of Treponema pallidum in an Arizona County with increasing syphilis morbidity: use of specimens from ulcers and blood.

A molecular-based subtyping system for Treponema pallidum was used during an investigation of increasing syphilis in Maricopa County, Arizona. Genital ulcer or whole blood specimens from patients with syphilis were assayed by a polymerase chain reaction (PCR) amplification of a T. pallidum DNA polymerase I gene. Positive specimens were typed on the basis of PCR amplification of 2 variable genes. In all, 41 (93%) of 44 of ulcer specimens and 4 (27%) of 15 blood specimens yielded typeable T. pallidum DNA. Twenty-four (53%) of 45 specimens were subtype 14f; other subtypes identified included 4f, 4i, 5f, 12a, 12f, 14a, 14d, 14e, and 14i. Only 2 specimens were from epidemiologically linked patients. This investigation demonstrates that multiple subtypes of T. pallidum can be found in an area with high syphilis morbidity, although 1 subtype (14f) was predominant. Four typeable specimens were from blood, a newly identified specimen source for subtyping.

Arizona↗
PubMed2004

Treponema pallidum strain-specific differences in neuroinvasion and clinical phenotype in a rabbit model.

BACKGROUND: The relationship between neuroinvasion and other manifestations of syphilis and the infecting strain of Treponema pallidum is not known. METHODS: Six groups of 8 rabbits were intravenously infected with 1 x 108 organisms from 1 of 6 strains of T. pallidum. Rabbits were examined 2-3 times/week; blood and cerebrospinal fluid (CSF) were collected weekly and every 2 weeks, respectively, for 10-12 weeks. Degree of CSF pleocytosis and skin-lesion severity were estimated by the area under the white blood cell-versus-time and lesion-versus-time curves. RESULTS: Maximum serum Venereal Disease Research Laboratory test titers, time to maximum titer, degree of CSF pleocytosis, and severity of skin lesions differed significantly among infecting strains. Overall, T. pallidum was identified, by reverse-transcriptase polymerase chain reaction, in CSF from 13 (27.7%) of 47 rabbits and was never identified in CSF from rabbits infected with 1 of the strains. The time course of detection varied by infecting strain. Severity of skin lesions and of CSF pleocytosis were inversely correlated (P=.005). CONCLUSIONS: There are particularly neuroinvasive T. pallidum strains, and the clinical phenotype of infection varies with infecting strain. This information could ultimately be used to identify patients at increased risk for neuroinvasion and, thus, at risk for neurosyphilis.

Animals↗
PubMed2006

Detection and discrimination of herpes simplex viruses, Haemophilus ducreyi, Treponema pallidum, and Calymmatobacterium (Klebsiella) granulomatis from genital ulcers.

BACKGROUND: Genital ulcer disease (GUD) is commonly caused by pathogens for which suitable therapies exist, but clinical and laboratory diagnoses may be problematic. This collaborative project was undertaken to address the need for a rapid, economical, and sensitive approach to the detection and diagnosis of GUD using noninvasive techniques to sample genital ulcers. METHODS: The genital ulcer disease multiplex polymerase chain reaction (GUMP) was developed as an inhouse nucleic acid amplification technique targeting serious causes of GUD, namely, herpes simplex viruses (HSVs), H. ducreyi, Treponema pallidum, and Klebsiella species. In addition, the GUMP assay included an endogenous internal control. Amplification products from GUMP were detected by enzyme linked amplicon hybridization assay (ELAHA). RESULTS: GUMP-ELAHA was sensitive and specific in detecting a target microbe in 34.3% of specimens, including 1 detection of HSV-1, three detections of HSV-2, and 18 detections of T. pallidum. No H. ducreyi has been detected in Australia since 1998, and none was detected here. No Calymmatobacterium (Klebsiella) granulomatis was detected in the study, but there were 3 detections during ongoing diagnostic use of GUMP-ELAHA in 2004 and 2005. The presence of C. granulomatis was confirmed by restriction enzyme digestion and nucleotide sequencing of the 16S rRNA gene for phylogenetic analysis. CONCLUSIONS: GUMP-ELAHA permitted comprehensive detection of common and rare causes of GUD and incorporated noninvasive sampling techniques. Data obtained by using GUMP-ELAHA will aid specific treatment of GUD and better define the prevalence of each microbe among at-risk populations with a view to the eradication of chancroid and donovanosis in Australia.

Calymmatobacterium↗
PubMed2006

Antibiotic selection may contribute to increases in macrolide-resistant Treponema pallidum.

To determine whether the 23S rRNA mutation that confers macrolide resistance is present in >1 Treponema pallidum strain, 58 isolates collected between 2001 and 2005 were screened for this mutation and for an unrelated sequence that distinguishes between strains. The odds of identifying a macrolide-resistant strain increased over time (P=.006). In subjects who had received macrolides in the previous year, the relative risk of harboring a resistant strain was 2.2 (95% confidence interval, 1.1-4.4; P=.02). The macrolide-resistant strains were not identical. These findings suggest that macrolide resistance may be increasing in multiple strains in response to antibiotic pressure.

Drug Resistance, Bacterial↗
PubMed1999

Cystalysin, a 46-kDa L-cysteine desulfhydrase from Treponema denticola: biochemical and biophysical characterization.

A 46-kDa hemolytic protein referred to as cystalysin, from Treponema denticola ATCC 35404, was characterized and overexpressed in Escherichia coli LC-67. Cystalysin lysed erythrocytes, hemoxidized hemoglobin to sulfhemoglobin and methemoglobin, and removed the sulfhydryl and amino group from selected S-containing compounds (e.g., cysteine) producing H2S, NH3, and pyruvate. With L-cysteine as substrate, cystalysin obeys Michaelis-Menten kinetics. Cystathionine and s-aminoethyl-L-cysteine were also substrates. Several of the small alpha amino acids were found to be competitive inhibitors of cystalysin. The enzymatic activity was increased by beta-mercaptoethanol and was not inhibited by the proteinase inhibitor TLCK (N alpha-p-tosyl-L-lysine chloromethyl ketone), pronase, or proteinase K, suggesting the functional site was physically protected or located in a small fragment of the polypeptide. We hypothesize that cystalysin is a pyridoxal-5-phosphate-containing enzyme with the activity of an alphaC-N and betaC-S lyase (cystathionase). Since high amounts of H2S have been reported in deep periodontal pockets, this metabolic enzyme from T. denticola may also function in vivo as an important virulence molecule.

Cystathionine gamma-Lyase↗
PubMed1998

Treponema pallidum, lipoproteins, and synthetic lipoprotein analogues induce human immunodeficiency virus type 1 gene expression in monocytes via NF-kappaB activation.

Syphilitic genital ulcers are cofactors for the bidirectional transmission of human immunodeficiency virus (HIV). U937 human promonocytic cells chronically infected with HIV-1 (U1 cells) or transiently transfected with wild type or mutant HIV long terminal repeat (LTR) reporter constructs were used to examine mechanisms that likely underlie Treponema pallidum-induced immune cell activation and consequent induction of HIV. Virulent T. pallidum, a representative native treponemal lipoprotein (NTp47), or synthetic lipoprotein analogues (lipopeptides) all induced HIV replication in U1 cells. These stimuli also induced HIV gene expression from a wild type HIV LTR. HIV gene expression correlated with the translocation of NF-kappaB, and mutations within the NF-kappaB binding sites of the HIV LTR abrogated HIV gene expression. This study implicates treponemal lipoproteins as key mediators of immune cell activation and provides insights into the cellular and molecular bases for enhanced HIV transmission in syphilitic persons.

Animals↗
PubMed1982

Cloning and expression of Treponema pallidum protein antigens in Escherichia coli.

Difficulties in culturing the bacterium Treponema pallidum have greatly hindered syphilis research. Recently, several laboratories have accomplished the expression of T. pallidum antigens in Escherichia coli. It is anticipated that treponemal antigens produced by recombinant DNA technology will provide new tools for investigating the pathogenesis and immunobiology of T. pallidum infection.

Antigens, Bacterial↗
PubMed1992

Treponema pallidum and Helicobacter pylori recovered in a case of chronic active gastritis.

A 49-year-old man complaining of epigastric pain underwent endoscopy, during which thickened stomach folds below the fundus were observed. Microscopic examination of gastric tissue biopsy specimens revealed chronic active gastritis. Dieterle stain revealed overwhelming numbers of "corkscrew-like" spirochetes. These were proved to be consistent with Treponema pallidum. A comprehensive study of the tissue revealed the added presence of Helicobacter pylori. This appears to be the first case report describing the involvement of H. pylori and T. pallidum together in a case of chronic active gastritis.

Chronic Disease↗
PubMedDate not supplied

Identification, cloning, and purification of protein antigens of Treponema pallidum.

Difficulties in culturing the bacterium Treponema pallidum have greatly hindered syphilis research. In recent years, several laboratories have begun applying recombinant DNA technology to the study of this organism. Recent work is summarized concerning the expression of T. pallidum DNA in Escherichia coli. A number of E. coli clones expressing treponemal protein antigens have been identified. In one instance, a recombinant protein was purified to homogeneity and shown to be identical to a highly immunogenic, native T. pallidum membrane protein of molecular weight 39,000, which was designated the basic membrane protein (BMP) of this organism. In addition, recent experiments are described that were designed to identify cell-surface proteins that would serve as the primary focus of our cloning efforts. Results obtained with use of several different approaches strongly suggest that the outer membrane of T. pallidum is an antigenically inert structure largely devoid of protein. However, a class of low-molecular-weight protein antigens have been identified that are actively secreted into the extracellular medium. Attempts currently are being made to clone these secreted proteins and investigate their roles in the pathogenesis and immunobiology of syphilis.

Animals↗
PubMed1993

Measurement of electrical bioimpedance for studying utilization of amino acids and peptides by Porphyromonas gingivalis, Fusobacterium nucleatum, and Treponema denticola.

The growth response of Porphyromonas gingivalis, Fusobacterium nucleatum, and Treponema denticola to peptides (supplied as trypticase) and amino acids (supplied as casamino acids) was measured over 24 hours by monitoring changes in alternating-current conductivity at 37 degrees C. All species utilized peptides preferentially over amino acids. These results are consistent with those obtained previously with conventional growth-response experiments. The duration of the linear growth response to trypticase of P. gingivalis was 9.7 hours, whereas that of both F. nucleatum and T. denticola was > 24 hours. By contrast, there was more uniformity in the utilization of amino acids from the casamino acid mixture, which previously has been shown to be a poor growth substrate. Furthermore, subtle differences in growth patterns, such as the ability of F. nucleatum to metabolize its storage glycopolymers before utilizing amino acids, were clearly evident. The present method provides an excellent means of studying bacterial growth kinetics and delineating bacterial/substrate specificities of both synthetic and natural substrates.

Amino Acids↗
PubMed1997

Congenital syphilis: detection of Treponema pallidum in stillborns.

Paraffin-embedded tissue from all 17 autopsies performed following 56 stillbirths associated with maternal syphilis during a 3-year period (1987-1989) was reexamined to compare immunofluorescent antigen (IFA) testing with silver staining for the detection of Treponema pallidum. Congenital syphilis (CS) originally was diagnosed in 9 of the 17 cases of stillbirth, on the basis of positive silver stains (7 cases) or morphological findings alone (2). Upon review, silver staining revealed T. pallidum in 10 of 17 cases and IFA testing revealed the pathogen in 15 of 17 cases, enabling diagnosis of CS in 16 of 17 cases of stillbirth associated with a reactive maternal rapid plasma reagin (RPR) card test. Most stillbirths associated with a reactive maternal RPR test during this time period involved CS, and IFA testing for T. pallidum is superior to silver staining for the identification of treponemes.

Adolescent↗
PubMed2000

Crystal structure of cystalysin from Treponema denticola: a pyridoxal 5'-phosphate-dependent protein acting as a haemolytic enzyme.

Cystalysin is a C(beta)-S(gamma) lyase from the oral pathogen Treponema denticola catabolyzing L-cysteine to produce pyruvate, ammonia and H(2)S. With its ability to induce cell lysis, cystalysin represents a new class of pyridoxal 5'-phosphate (PLP)-dependent virulence factors. The crystal structure of cystalysin was solved at 1.9 A resolution and revealed a folding and quaternary arrangement similar to aminotransferases. Based on the active site architecture, a detailed catalytic mechanism is proposed for the catabolism of S-containing amino acid substrates yielding H(2)S and cysteine persulfide. Since no homologies were observed with known haemolysins the cytotoxicity of cystalysin is attributed to this chemical reaction. Analysis of the cystalysin-L-aminoethoxyvinylglycine (AVG) complex revealed a 'dead end' ketimine PLP derivative, resulting in a total loss of enzyme activity. Cystalysin represents an essential factor of adult periodontitis, therefore the structure of the cystalysin-AVG complex may provide the chemical basis for rational drug design.

Binding Sites↗
PubMed1976

Relative efficacy of clindamycin, erythromycin, and penicillin in treatment of Treponema pallidum in skin syphilomas of rabbits.

The currently recommended antibiotic for treatment of fetal syphilis in pregnant women who are allergic to penicillin is erythromycin. However, clindamycin crosses the placenta more effectively than erythromycin. Therefore, an in vivo rabbit model of intradermal syphilomas was used to determine the effect of clindamycin compared with the effects of erythromycin and penicillin on the growth of virulent Treponema pallidum. The average number of motile treponemes in two, paired, mature lesions was determined before and after therapy in groups of four rabbits per dosage. Single intramuscular doses of clindamycin (15 and 40 mg/kg) and erythromycin (12 and 40 mg/kg) did not decrease treponeme counts significantly. Single injections of penicillin (10,000 units/kg) reduced treponemal counts by more than 250-fold. Multiple intramuscular injections of clindamycin reduced counts by five- to sevenfold, whereas multiple doses of erythromycin and penicillin decreased treponeme counts by greater than 300-fold. These studies indicate that clindamycin is far less active than erythromycin or penicillin in treatment of established syphilitic lesions in rabbits.

Animals↗
PubMed1978

Detection by direct immunofluorescence of antibodies to Treponema pallidum in the cutaneous infiltrates of rabbit syphilomas.

By direct immunofluorescence, with use of fluorescein-labeled sonified Treponema pallidum, specific antibodies were detected in the tissue infiltrates of cutaneous syphilomas in rabbits. Specimens from cutaneous and mucosal inflammatory lesions induced by intradermal injection of a keratinous substance from an epidermal cyst served as controls. Granular fluorescence was detected in the dermis of 11 of the 12 syphiloma specimens and corresponded to areas of heavy plasma cell infiltrates, and some fluorescence was found directly on plasma cells identified by subsequent staining with hematoxylin and eosin. This fluorescence could be blocked by preabsorption. Control slides did not show any fluorescence. Immunopathologic techniques using labeled antigens may be of diagnostic value in syphilis and other infectious disorders which feature specific infiltrates.

Animals↗
PubMed1980

Endemic syphilis: transfer of resistance to Treponema pallidum strain Bosnia A in hamsters with a cell suspension enriched in thymus-derived cells.

Direct evidence for involvement of thymus-derived (T) cells in host defense against syphilitic infection is presented. Irradiated, bone marrow-reconstituted hamsters receiving cells from hamsters immune to infection with Treponema pallidum strain Bosnia A had significantly lower weights and fewer treponemes per lymph node than animals that had received normal lymphoid cells or only normal bone marrow cells. Cell suspensions enriched in T cells from immune hamsters were obtained by sequential filtration of pooled spleen and lymph node cells through glass- and nylon-wool columns. The fractionated suspensions of cells responded poorly to stimulation by phytohemagglutinin, Escherichia coli lipopolysaccharide, and dextran sulfate, but responded relatively strongly to the T-cell mitogen, convanavalin A. After fractionation the proportion of cells susceptible to antithymocyte serum and complement increased significantly. These immune cell suspensions depleted of bone marrow-derived cells closely resembled unfractionated suspensions in their ability to confer resistance to challenge with T. pallidium.

Animals↗
PubMed1983

The interaction between Treponema pallidum and human polymorphonuclear leukocytes.

The interaction between polymorphonuclear leukocytes (PMNLs) and Treponema pallidum was studied. Intradermal injection of greater than or equal to 10(6) T. pallidum into rabbits caused a rapid accumulation of PMNLs. Human serum released chemotaxigenic factor (C5a) during incubation in vitro with T. pallidum. Incubation of T. pallidum with human PMNLs in vitro (ratio, 100:1) stimulated chemiluminescence. These responses were dependent upon the presence of both antibody and complement and were greatest when serum from a patient with late secondary syphilis was used as a chemotaxigenic source or for opsonization. Electron microscopic studies documented the rapid uptake of T. pallidum into membrane-bound vacuoles in the human PMNLs in vitro after incubation for as little as 5 min, with leukocyte degranulation and loss of treponemal integrity observed after 4 hr. T. pallidum were found within PMNLs 3 hr after intradermal inoculation of rabbits. These data show that PMNLs are attracted to, and appear to ingest, T. pallidum, but they fail to explain why inoculation of these organisms is not followed by eradication.

Animals↗
PubMed1985

Antigens of Treponema pallidum recognized by IgG and IgM antibodies during syphilis in humans.

IgG and IgM antibody specificities for antigens of Treponema pallidum Nichols strain were determined by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the western blot technique in sera from patients with untreated syphilis, normal persons, persons with biologic false-positive tests for syphilis, and sexual contacts of persons with infectious syphilis. IgG reactivities of sera from individuals with primary syphilis varied considerably but consistently exhibited strong reactivity to a 48-kilodalton band. Sera from patients with secondary and early latent syphilis uniformly demonstrated reactivity to 22 separate polypeptide antigens; decreased reactivity was seen in late latent syphilis. Normal and biologic false-positive sera showed weak IgG reactivity against none to 12 polypeptides. Sera from asymptomatic contacts of persons with infectious syphilis showed reactivity to a varying number of treponemal antigens, including some reactions not seen with normal sera. IgM reactivity was most prominent in secondary syphilis but was demonstrable at all stages of disease.

Antibodies, Bacterial↗