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PubMed2005

Defining the interaction of the Treponema pallidum adhesin Tp0751 with laminin.

Various invasive pathogens attach to host tissues via the extracellular matrix component laminin, the major glycoprotein found within basement membranes. Previous investigations identified the laminin-binding adhesin Tp0751 within the spirochete bacterium Treponema pallidum. In the current study, Tp0751 was shown to attach to a variety of laminin isoforms that are widely distributed throughout the host, including laminins 1, 2, 4, 8, and 10. Such universal attachment is conducive for an adhesin present within a highly invasive pathogen that encounters a variety of tissue sites during the course of infection. Additional studies systematically identified the amino acid residues within Tp0751 that contribute to laminin binding using synthetic peptides designed from the mature protein sequence. The minimum laminin-binding region of the adhesin was localized to 10 amino acids; peptides containing these residues inhibited attachment of Tp0751 and T. pallidum to laminin. Further, Tp0751-specific antibodies inhibited attachment of T. pallidum to laminin. This study furthers our knowledge of the interaction of T. pallidum with laminin, an association that is proposed to facilitate bacterial traversal of basement membranes and subsequent entry into the circulation and tissue invasion. As such, these investigations will reveal new targets for possible prevention of bacterial dissemination and establishment of chronic infection.

Adhesins, Bacterial↗
PubMed1999

Trends and interaction of HIV-1 and HIV-2 in Guinea-Bissau, west Africa: no protection of HIV-2 against HIV-1 infection.

OBJECTIVES: To study trends in the prevalence and incidence of HIV-1 and HIV-2 infections in Guinea-Bissau over the last 7 years, and to evaluate the protective effect of HIV-2 against HIV-1 infection. DESIGN: Prospective follow-up of a cohort of police officers in Guinea-Bissau, and sentinel surveillance of pregnant women in Bissau. METHODS: Participants in the police cohort were tested regularly for antibodies to HIV and Treponema pallidum, and information about sexual risk behaviour and a history of sexually transmitted diseases was obtained. Simultaneously, pregnant women at the maternity wards at the National Hospital in Bissau were screened annually for HIV antibodies. To evaluate changes in prevalence and incidence of HIV in the police cohort, the study period was divided into three time strata with 2-3 years in each stratum. For the evaluation of a protective effect of HIV-2 on subsequent HIV-1 infection, two multivariate Poisson regression models were constructed, adjusting for different selected confounding variables. RESULTS: Between 1990 and 1997, 2637 police officers were included in the cohort study, 90.7% of whom were male. The overall prevalence of HIV-1 was 0.9%, of HIV-2 it was 9.7% and of HIV-1 and HIV-2 dual reactivity it was 0.5%. For pregnant women the prevalence rates were 0.9, 5.5 and 0.2% for HIV-1, HIV-2 and dual reactivity respectively. The prevalence of HIV-1 increased significantly whereas the prevalence of HIV-2 declined significantly during the study period, among both police officers and pregnant women. The total incidence of HIV-1 and HIV-2 was 0.74 and 0.83 per 100 person-years respectively in the police cohort. The incidence of HIV-1 increased slightly from 0.62 to 0.78 per 100 person-years (not significant), whereas the incidence of HIV-2 declined significantly from 0.90 to 0.35 per 100 person-years over the study period. Seven police officers seroconverted from HIV-2 to dual reactivity (1.22 per 100 person-years). The adjusted incidence ratio of acquiring HIV-1 infection among HIV-2-positive subjects compared with HIV-negative subjects was 1.65 [95% confidence interval (CI), 0.73-3.74] and 1.98 (95% CI, 0.80-4.87), depending on the confounding variables included. CONCLUSIONS: Our study shows an increasing prevalence of HIV-1 and a decreasing prevalence of HIV-2 in Guinea-Bissau. The incidence of HIV-2 declined significantly whereas the incidence of HIV-1 was relatively stable over the study period. No protective effect of HIV-2 against subsequent HIV-1 infection was observed, instead HIV-2-positive subjects had a tendency towards higher risk of acquiring HIV-1 infection compared with seronegative subjects.

Africa↗
PubMed1985

Immunodiagnostic test for detection of serum antibody to Treponema pallidum (syphilis): fibronectin as a capture vehicle for treponemal adhesins.

Knowledge that Treponema pallidum adhesin proteins bind to host fibronectin (Fn) via ligand-receptor interactions has resulted in development of an ELISA for measuring specific antitreponemal antibodies in sera of syphilitic patients and infected experimental animals. As little as 50 ng of T. pallidum total protein extract added to Fn-coated wells permitted half-maximal levels of ELISA reactivity. Detection of serum antibody from intratesticularly infected rabbits occurred at dilutions greater than 1/100,000. Antibody titers in serum from patients with primary and latent syphilis were positive at 1/1 000 dilutions while serum samples from patients with secondary syphilis were reactive at 1/10,000. Furthermore, the ELISA proved useful for evaluating serum samples from individuals with other treponemal infections. Antibodies raised against the non-pathogenic spirochete, T. phagedenis biotype Reiter, were non-reactive with Fn-T. pallidum complexes. Also, Reiter treponemal proteins did not bind to Fn-coated wells. This ELISA using Fn as a capture vehicle for treponemal adhesin proteins was superior to 3 other routinely used tests for syphilis diagnosis.

Adhesins, Bacterial↗
PubMed1989

[Comparison of methods for the demonstration of Treponema-specific IgM].

In this research 113 sera have been analysed with three methods for IgM treponema-specific determination: IgM-SPHA, IgM-EIA and 19S IgM FTA-ABS. Among these sera, 33 samples related to non-treated patients at different stages of infection, and 80 samples to treated patients. The results point out a light sensibility of the IgM-SPHA in primary lues. The IgM-EIA test has performed a good specificity, but displayed a sensibility lower to the 19S IgM FTA-ABS, which proved the best of tests. About the cases of treated lues at different stages, in 23 samples with VDRL negative has been found no positivity at three tests used, while in 49 samples with VDRL positive 8 are resulted positive at 19S IgM FTA-ABS.

Antibodies, Bacterial↗
PubMed1987

Effect of early and late syphilis on central nervous system: cerebrospinal fluid changes and neurological deficit.

Neurological examination and investigation of the cerebrospinal fluid (CSF) was performed on 24 patients with early and 180 patients with late syphilis. In 21 (12%) patients with late syphilis positive CSF treponemal test results and neurological deficits suggestive of symptomatic neurosyphilis were found. Concomitantly all but three patients with neurosyphilis showed one or more of the following abnormal CSF variables: CSF concentration of albumin X 10(3)/serum concentration (albumin ratio) greater than or equal to 7.9; mononuclear cells greater than 5 microliters: ratio of CSF to serum IgG concentrations/ratio of CSF to serum albumin concentrations (IgG index) greater than or equal to 0.7 or of IgM/albumin (IgM index) greater than or equal to 0.1; or oligoclonal CSF immunoglobulins. In 20 (95%) patients with neurosyphilis evidence of the production of treponemal antibodies within the central nervous system (CNS) was shown. Ten (48%) patients with neurosyphilis had been treated previously for late syphilis. These observations emphasise the need to screen for neurosyphilis in patients with late syphilis. Intrathecal production of treponemal antibodies was detected in six (25%) patients with early and 44 (28%) with late syphilis who did not show any neurological deficit. Intrathecal production of treponemal antibodies indicating that the CNS was affected led us to suspect asymptomatic neurosyphilis in these patients. Seventeen (11%) patients with late syphilis but no neurosyphilis and only one (4%) with early syphilis showed additional abnormal CSF variables. Surprisingly, six out of 22 patients with treated early and 20 out of 68 patients with treated late syphilis showed evidence of treponema antibody production within the CNS. We do not know whether these findings indicate that the CNS was affected because of inadequate treatment or merely reflect persistent synthesis of treponemal antibodies associated with cured infection. In one (4%) patient with early and in 21 (13%) with late syphilis but no neurosyphilis abnormal CSF variables in the absence of positive CSF treponemal test results were observed, which excluded syphilitic inflammation of the CNS.

Antibodies, Bacterial↗
PubMed2000

Immunoperoxidase technique for detecting spirochetes in tissue sections: comparison with other methods.

BACKGROUND: With the increasing incidence of human immunodeficiency virus (HIV) infection and immunosuppressive therapy, the incidence of syphilis has been increasing. Given the fact that the above conditions may mask or obscure the usual clinical signs and symptoms of syphilis, a means of enhanced detection is essential. AIMS METHODS: The purpose of this study was to determine whether an immunoperoxidase method using an antibody against treponemes would increase the sensitivity and specificity of diagnosis in biopsies of patients with secondary syphilis. This was compared to serology and silver stain in cases of known syphilis. RESULTS: Immunoperoxidase for treponemes was at least as sensitive (9/10) as pathology (9/10), and more sensitive than conventional silver stain (6/10) or serology (7/10). CONCLUSIONS: In those equivocal cases of secondary syphilis, where confirmation is essential, immunoperoxidase for treponemes may be a useful adjunct.

AIDS-Related Opportunistic Infections↗
PubMed2000

Posterior uveitis in patients with positive serology for syphilis.

The clinical features and ophthalmologic findings of 20 patients with syphilitic posterior uveitis seen at the Detroit Medical Center from November 1993 through February 1996 were reviewed. The mean age was 58 years; 8 patients were male and 12 were female; and all patients were black. Three of 9 patients tested were HIV positive. Patients were divided into 2 groups: those with acute (8) and those with chronic (12) syphilitic posterior uveitis. Chorioretinitis was the predominant uveitic pattern (15/20). Eighteen patients presented with blurred vision. All patients had reactive serum fluorescent treponemal antibody, absorbed (FTA-ABS); 3 had nonreactive rapid plasma reagin (RPR). Mean RPR titer in the chronic uveitis group and in the acute uveitis group was 1:27.3 and 1:209.8, respectively. Seven patients had abnormal cerebrospinal fluid (CSF); CSF VDRL was reactive in 2 patients. All patients were treated with intravenous penicillin G. Eight of 14 patients seen at follow-up showed improvement of ophthalmologic findings. Syphilis should be considered in the differential diagnosis of posterior uveitis.

Adult↗
PubMed1998

Molecular subtyping of Treponema pallidum subspecies pallidum.

BACKGROUND AND OBJECTIVES: Epidemiologic studies on syphilis have been hampered by the fact that strains of Treponema pallidum subspecies pallidum (T. pallidum), the causative agent of this disease, cannot be differentiated by either protein-based or deoxyribonucleic acid-based methods. Syphilis is endemic in many developing countries and is common in some industrialized nations. In addition, the disease has been shown to increase the risk of infection with the human immunodeficiency virus. GOAL: To develop a molecular subtyping method for T. pallidum. STUDY DESIGN: Two genes exhibiting intrastrain variability were identified as potential targets for strain differentiation: the acidic repeat protein (arp) gene, which contains a variable number of 60 base pair repeats, and a member of the treponema pallidum repeat (tpr) gene family. Polymerase chain reaction amplification and restriction endonuclease digestion of polymerase chain reaction products from laboratory strains and clinical specimens were used to develop a molecular subtyping scheme for T. pallidum. RESULTS: Determining the number of repeats in the arp gene by polymerase chain reaction resulted in 12 different subtypes among the 63 isolates that were studied. Among those, most (54.2%) had arp genes with 14 repeats. The other 11 subtypes had arp genes with 7 to 21 repeats, each accounting for 2% to 14% of the isolates. Polymerase chain reaction amplification of a member of the tpr gene family from a subset of 46 isolates followed by digestion of the polymerase chain reaction product with MseI resulted in seven restriction fragment length polymorphism patterns designated a to g. Strains with 14 repeats could be grouped into five restriction fragment length polymorphism subtypes. By combining the two systems we observed 16 subtypes among 46 isolates examined. This typing system is stable, reproducible, and easy to perform. In addition, the use of the ABI Genetic Analyzer for the determination of fragment size and banding patterns makes the results unbiased. CONCLUSION: This is the first molecular subtyping system that distinguishes among clinical isolates of T. pallidum.

Animals↗
PubMed1999

[Immunoblotting for the serodiagnosis of syphilis. A candidate to replace the Nelson-Mayer test].

The lipid antigens used in the Bordet-Wassermann and prepared by Landsteiner and Marie for syphilis tests from 1909 to 1949 were non specific but have certain common features with the spirochete body. For more than forty years the Bordet-Wassermann reaction, associated with flocculation (Kahn) or agglutination (Kline-VDRL) was used to detect cases of tremonematosis despite frequent false positives reactions due to other infections. In 1949, the Nelson and Mayer test was introduced. This test was based on a rigorously specific reaction based on an antigen of live virulent Treponema pallidum. Culture being impossible, the strain had to be, and still is, maintained by weekly passage on rabbit testicles. These manipulations are very dangerous and the technique is difficult, being performed only in specialized laboratories. This test however made it possible to identify the specificity of lipid tests and led to the development of specific immunofluorescent reactions (FTA) in 1959 and of hemagglutination test (TPHA) in 1969. In 1980, we introduced a simple treponemic reaction (FTA or TPHA) associated with a lipid reaction (VDRL) for screening. The specificity of these tests is not however perfect and the Nelson test remains useful as a highly specific reaction. A simple test with comparable specificity was long awaited and is now available with immunoblotting as for HIV, boreliosis and pertussis, etc. We propose this new reaction to replace the Nelson test because it is specific, is sensitive early, distinguishes between IgG and IgM and is not dangerous to manipulate. We have tested it in over one hundred selected sera of CSF from subjects with recent, former or nervous syphilis as well as cases susceptible of producing false positive reactions and have concluded that immunoblotting is highly specific and sensitive. We recommend official approval of this test to replace the Nelson test.

Humans↗
PubMed1974

Intrauterine infection and cord immunoglobulin M. 3. Serological analysis of infants with elevated cord serum immunoglobulin M.

The presence of antibodies to rubella, cytomegalovirus and Toxoplasma gondii was determined at birth and at 6 months of age in a group of 147 infants with cord serum IgM levels >/= 19.0 mg/dl and in 92 control infants. Maternal syphilis serology was determined in both groups as well. No significant differences in the prevalence or levels of antibodies to these pathogens were found between the two groups which might have led to the diagnosis of unsuspected intrauterine infection. Persistence of antibodies to 6 months of age was similar in the two groups, indicating that this is not a useful index of intrauterine infection.ANALYSIS OF THE RESULTS YIELDED THE FOLLOWING DATA ON THE PREVALENCE OF ANTIBODIES TO THE PATHOGENS STUDIED: rubella virus, 90 and 75% seropositivity at birth and 6 months respectively; cytomegalovirus, 65 and 35%; and Toxoplasma gondii, 33% seropositivity at birth.

Antibodies↗
PubMed1989

Major integral membrane protein immunogens of Treponema pallidum are proteolipids.

A number of the major pathogen-specific immunogens of Treponema pallidum were characterized recently as amphiphilic, integral membrane proteins by phase partitioning with Triton X-114 (J. D. Radolf, N. R. Chamberlain, A. Clausell, and M. V. Norgard. Infect. Immun. 56:490-498, 1988). In the present study, we demonstrated that the same membrane immunogens (designated as detergent phase proteins [DPPs]) become radiolabeled upon in vitro incubation of T. pallidum with various 3H-labeled fatty acids. Radioimmunoprecipitation with a monoclonal antibody confirmed that the 3H-labeled 47-kilodalton protein corresponded to the well-characterized treponemal antigen with the identical apparent molecular mass. Failure to detect 3H-labeled DPPs following incubation with erythromycin confirmed that protein acylation required de novo protein synthesis by the bacteria. When treponemes were incubated with [3H]myristate, [3H]palmitate, or [3H]oleate, radiolabeled proteins corresponding to the DPPs were detected upon autoradiography. Demonstration that a number of the abundant membrane immunogens of T. pallidum are proteolipids provides information to help clarify their membrane association(s) and may serve to explain their extraordinary immunogenicity.

Animals↗
PubMedDate not supplied

Occurrence of congenital, cardiovascular, visceral, neurologic, and neuro-ophthalmologic complications in late yaws: a theme for future research.

Late lesions of yaws are thought to be limited to skin, bones and joints, without congenital, visceral, or central nervous system (CNS) involvement. However, the treponemes isolated from patients with yaws, endemic syphilis, and venereal syphilis are genetically identical subspecies of Treponema pallidum. The controversy surrounding distinctions between syphilis and yaws is presented in its historical context, and reports of congenital, visceral, and CNS complications of yaws are reviewed. Isolation of treponemes from aqueous humor, as well as CSF abnormalities in 24.9% of 902 patients with yaws, indicate the existence of CNS involvement. The high prevalence of tropical myeloneuropathies of unknown origin discovered in areas of previous treponemal endemicity, particularly in Jamaica and Colombia, may represent late complications of yaws. Careful analysis of the collected evidence indicates that potential sequelae of yaws include congenital, visceral, and tertiary CNS lesions identical to those of venereal syphilis. The current worldwide resurgence of endemic treponematoses provides an unparalleled opportunity to settle conclusively questions still unanswered regarding the natural history of these infections.

Cardiovascular Diseases↗
PubMed1986

Effect of immune serum and its immunoglobulin fractions on hamsters challenged with Treponema pallidum ssp. pertenue.

Passive transfer of frambesial immune serum is capable of conferring complete protection on hamsters against challenge with Treponema pallidum ssp. pertenue. Treponemicidal activity in the pooled immune serum is relatively high. Immune serum and its immunoglobulin fractions, especially IgG2, also killed T. pallidum ssp. pertenue in vitro. Treponemicidal activity was present only when immune serum was administered to hamsters within a short time (three days) of frambesial challenge. By contrast, administration of pooled immune serum to hamsters infected for more than one week failed to reduce the number and size of lesions and the weight and number of treponemes in the lymph nodes. These results suggest that hamsters can develop the humoral components necessary to protect them against frambesial challenge, but these components are unable to destroy treponemes at the primary foci of infection.

Animals↗
PubMed1999

Vertical transmission of Treponema pallidum to various litters and generations of guinea pigs.

The transmission of congenital syphilis was studied in a 4-generation guinea pig family with 10 litters and 38 offspring. By use of one or all of the following tests (ELISA-IgM, polymerase chain reaction, and rabbit infectivity), transplacental infection was demonstrated through 5 litters and up to 4 generations. Twenty-eight (93%) of 30 animals were positive by >/=1 test, and 2 (7%) were negative by 1 or 3 tests. While transmission of the pathogen appeared to be unaffected by the maternal acquisition of immunity, signs of smoldering infection in the young was suggested by the decline in humoral responses in successive progeny and by unusual rabbit infectivity test results. With each pregnancy there was a remarkable booster in the maternal humoral response, which dropped significantly prior to term. These findings shed new light on the understanding and interpretation of serologic testing during pregnancy and the perinatal period.

Animals↗
PubMedDate not supplied

Anaerobic oral and dental infection.

Anaerobes make up a significant part of the oral and dental indigenous and pathogenic flora. Their role in periodontal disease, root canal infections, infections of the hard and soft oral tissue, as well as their importance as foci for disseminated infectious disease is well established. Despite the ubiquitous involvement of bacteria, significant progress in our understanding of specific microbial etiologies has occurred only in the past decade. Estimates of the number of species recovered from samples of subgingival plaque range from 250 to 400, a large portion made up by anaerobes. Common anaerobic isolates include Fusobacterium, Bacteroides, Actinomyces, Peptococcus, Peptostreptococcus, Selenomonas, Eubacterium, Propionibacterium, and Treponema. Recently, several significant advances in our knowledge have set the stage for future research. First, circulating levels of hormones in pregnant women were shown to be stimulatory to Bacteroides species, which were associated with increased levels of gingival infection. Second, bacterial invasion of the soft and hard periodontal tissues has been documented in gingivitis, advanced periodontitis, and localized juvenile periodontitis. The frequency and identity of invading bacteria will determine the implications for diagnosis and treatment. Third, antibacterial "probes" aimed at anaerobic (and capnophilic) bacteria have had promising results in controlling and arresting oral, dental, and peridontal anaerobic infections.

Anti-Bacterial Agents↗
PubMed1998

A new recombinant antigen latex agglutination test (Syphilis Fast) for the rapid serological diagnosis of syphilis.

We report an assessment of Syphilis Fast, a new latex test that uses a pool of 3 recombinant Treponema pallidum antigens (TpN15, TpN17, and TpN47) for the serodiagnosis of syphilis. Specificity was evaluated by screening 1518 unselected blood specimens in parallel with Syphilis Fast, the Captia SelectSyph-G EIA and the Venereal Disease Research Laboratory (VDRL) cardiolipin antigen test while sensitivity was tested using a panel of 99 treponemal sera (treated and untreated) representing various stages of infection and 15 treponemal sera detected on screening. The specificity of Syphilis Fast on initial testing (99.8%) was significantly higher (P<0.02) than that of Captia SelectSyph-G (99.2%) and the VDRL (99.1%): the specificity of Syphilis Fast remained significantly higher (P<0.02) after repeat testing (respective values 99.9%, 99.5% and 99.4%). There was no difference in the sensitivity of Syphilis Fast and Captia SelectSyph-G on initial (93% vs 92.1%) or repeat (95.6% vs 94.7%) testing: both were significantly more sensitive (P<0.001) than the VDRL (46.5% on initial and 43.9% on repeat testing). The sensitivities of the Treponema pallidum haemagglutination test (TPHA) and FTA-abs were 98.2% and 95.6% respectively. Negative reactions in Syphilis Fast and SelectSyph-G were associated with treated infections and correlated with low TPHA titres (< or = 80). We conclude that Syphilis Fast is a highly specific, simple and fast screening test with a sensitivity comparable to native antigen treponemal tests and that it merits consideration as a front-line screening test.

Antibodies, Bacterial↗
PubMed1989

Serum lipoprotein binding by Treponema pallidum: possible role for proteoglycans.

Acquisition by the syphilis spirochaete, Treponema pallidum, of radioiodinated total human plasma lipoprotein and lipoprotein subfractions was examined. Time dependent and saturation binding kinetics were observed for total lipoproteins and subfractions, including high density lipoproteins, low density lipoproteins (LDL), and very low density lipoproteins. All subfractions competed equally well in binding iodinated total lipoproteins and individual subfractions, but apoproteins common to all subfractions were ineffective in inhibiting lipoprotein acquisition. The interaction of LDL with T pallidum was studied further and, interestingly, the presence of 17% sulphated dextran sulphate (DS) in the reaction mixture containing treponemes and LDL resulted in up to 172 times more LDL being bound by live treponemes. Biological variability was observed in the extent of increased LDL bound in the presence of 17% sulphated DS by preparations of T pallidum isolated from different infected rabbits. Saturation kinetics of iodinated LDL acquisition was obtained in the presence of 17% sulphated DS but not 1% sulphated DS. Other proteoglycan molecules, such as chondroitin sulphate, hyaluronic acid and heparin, and fibronectin, the extracellular matrix protein targeted by treponemes in parasitism of host cells and tissues neither diminished nor enhanced LDL binding by live treponemes. Only 5% and 10% of associated radioactivity was released from treponemal surfaces after T pallidum was incubated with iodinated LDL and 17% sulphated-DS for 15 and 30 minutes, respectively. These data show binding and possible internalisation of host lipoproteins by T pallidum, which may be mediated by sulphated proteoglycan. Sulphated proteoglycans accumulate during T pallidum infections of host cells.

Dextran Sulfate↗