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PubMed2006

Quantitative analysis of tpr gene expression in Treponema pallidum isolates: Differences among isolates and correlation with T-cell responsiveness in experimental syphilis.

Transcriptional analysis of the tpr genes in Treponema pallidum subsp. pallidum (referred to here as simply T. pallidum) has been limited to date, and yet the expression of members of this gene family is likely relevant to the pathogenesis of syphilis. Recently, immunological studies and semiquantitative mRNA analysis led to the hypothesis of the modulation of tpr gene transcription during infection and suggested that various strains of T. pallidum might differentially express these genes. In this study we developed a real-time amplification assay to quantify the tpr mRNAs with respect to the 47-kDa lipoprotein message and to compare transcript levels among four different strains of T. pallidum. In addition, we analyzed the lymphocyte responsiveness pattern toward the Tpr antigens in late experimental syphilis to identify tpr genes that had been expressed during the course of infection. The T-cell response has been implicated in clearance of treponemes from early lesions, and some of the Tprs were identified as strong targets of the cellular immune response. We show that message for many of the tpr genes can be detected in treponemes harvested at the peak of early infection. Interestingly, tprK seems to be preferentially expressed in almost every strain, and it is uniformly the target of the strongest cellular immune response. These studies demonstrate the differential expression of certain tpr genes among strains of T. pallidum, and further studies are needed to explore the relationship between tpr gene expression and the clinical course of syphilis in infected individuals.

Animals↗
PubMed1975

Lymphocyte transformation in syphilis: an in vitro correlate of immune suppression in vivo?

Suppression of cellular immunity during primary and secondary infection may explain, in part, the unusual clinical evolution of syphilis. We have previously shown that lymphocytes from normal subjects undergo blastic transformation when exposed in vitro to Treponema refringens. This response was suppressed in patients with syphilis. the suppression being unrelated to serum factors. In the present paper we studied lymphocyte response in vitro to T. refringens, T. reiter, and T. pallidum as well as to monilia and trychophytins. The response to these antigens was suppressed in patients with syphilis although the response to phytohemagglutinin. pokeweed mitogen, and streptolysin was normal. These data support the hypothesis that human infection with T. pallidum is followed by a complex interaction between cellular and humoral immunity, the former being suppressed in primary and secondary stages.

Antigens, Bacterial↗
PubMed1984

Molecular analysis of immunoglobulins M and G immune response to protein antigens of Treponema pallidum in human syphilis.

Protein antigens of Treponema pallidum precipitated by immunoglobulin M (IgM) and IgG antibodies of sera from patients with untreated primary and secondary syphilis as well as treated secondary syphilis were characterized on a molecular basis. T. pallidum was labeled internally with [35S]methionine and solubilized in 0.1% sodium dodecyl sulfate-1% Triton X-100. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis on 12.5% gels followed by autoradiography revealed 32 distinct proteins with molecular weights between 13,500 and 200,000. Twenty-three proteins of T. pallidum with molecular weights between 15,500 and 115,000 were identified as antigens by double antibody radioimmunoprecipitation with IgM and IgG antibodies of sera from syphilitic patients. The molecular analysis of the IgM and IgG immune response to T. pallidum in human syphilis is in accord with earlier immunological observations. Finally, utilizing syphilitic human sera, we characterized 15 protein antigens of T. pallidum that are common to Treponema phagedenis by partial absorption of IgM and IgG antibodies with an ultrasonicate of T. phagedenis.

Antibodies↗
PubMed1999

T-Cell responses to Treponema pallidum subsp. pallidum antigens during the course of experimental syphilis infection.

In this study we describe the development of the T-cell response to a panel of Treponema pallidum antigens over the course of syphilis infection in the rabbit and determine whether these antigens induce the expression of Th1 cytokines. It was determined that the membrane proteins TpN17 and TpN47, as well as the endoflagellar sheath protein TpN37, induce strong proliferation responses through most of syphilis infection; Tromp1 induced only weak proliferative responses. An unexpected drop in proliferative response to these antigens at day 90 of infection, followed by a dramatic increase in response at day 180, suggests that there may be a secondary dissemination of T. pallidum which induces a recall response. Crude epitope mapping of TpN17 and TpN37 showed that multiple epitopes may be present on both antigens, which is likely a contributing factor in the immunodominance of these antigens. The T-cell response to the TpN37 molecule shows acquisition of newly recognized epitopes during the course of infection. Sonicated T. pallidum was found to induce the expression of interleukin-2 (IL-2) and gamma interferon and not IL-10 mRNA, showing that the general T-cell response to T. pallidum antigens in syphilis infection is biased towards the Th1 phenotype. Of the antigens tested, TpN37 appears to contribute the most to the Th1 cytokine response and therefore may play a key role in the clearance of T. pallidum from lesions.

Animals↗
PubMed1974

In vitro lymphocyte response to Treponema refringens im human syphilis.

The response of lymphocytes from patients with syphilis and normal subjects was studied in vitro by using phytohemagglutinin (PHA), pokeweed mitogen (PWM), streptolysin O (SLO), and a preparation of Treponema refringens. Normal lymphocytes exhibited a dose-response curve to treponemes. Although lymphocytes from patients with primary and secondary syphilis responded normally to PHA and PWM, their response to SLO was suppressed and they failed to show significant stimulation by treponemes. Serum from syphilitic patients did not affect normal lymphocytes, and culturing lymphocytes from patients with syphilis in normal serum did not restore their responsiveness. Six to 10 weeks after syphilitic patients had been treated, the degree of stimulation by treponemes was the same as for normal subjects. These data give indirect support to the hypothesis that immunological suppression occurs during active infection with T. pallidum.

Antigens, Bacterial↗
PubMed1981

Specificity, sensitivity, and reproducibility among the fluorescent treponemal antibody-absorption test, the microhemagglutination assay for Treponema pallidum antibodies, and the hemagglutination treponemal test for syphilis.

Using 920 sera, we compared the specificity and reproducibility of the hemagglutination treponemal test for syphilis with those of the fluorescent treponemal antibody-absorption test and the microhemagglutination assay for Treponema pallidum antibodies; we found all three tests to be comparable. However, the hemagglutination treponemal test for syphilis, like the microhemagglutination assay for T. pallidum antibodies, lacked sensitivity in sera from patients with primary syphilis.

Antibodies, Bacterial↗
PubMed1982

Heated versus unheated sera in the hemagglutination treponemal test for syphilis.

Sera (920) were tested to evaluate the use of heated versus unheated sera in the hemagglutination treponemal test for syphilis. The heated and unheated samples were tested on the same day with the hemagglutination treponemal test for syphilis kit according to the manufacturer's protocol. Agreement of results between the heated and unheated sera was 99.2%. The reading pattern of agglutination was clearer and more distinct with heated sera; therefore, based solely on our preference for the reading patterns, we suggest that heated sera be used in the hemagglutination treponemal test for syphilis.

Hemagglutination Tests↗
PubMed1983

Reactivity of microhemagglutination, fluorescent treponemal antibody absorption, Venereal Disease Research Laboratory, and rapid plasma reagin tests in primary syphilis.

Seroreactivity of sera from 109 patients with first-infection primary syphilis was 98.2% in the fluorescent treponemal antibody absorption test, 92.7% in the rapid plasma reagin 18-mm circle card test, 72.5% in the microhemagglutination test (MHA-TP), and 72.5% in the Venereal Disease Research Laboratory test. Seroreactivity of sera from 18 patients with primary syphilis with documented previous infection(s) was 100% in the fluorescent treponemal antibody absorption test, the rapid plasma reagin 18-mm circle card test, and the MHA-TP test and 88.9% in the Venereal Disease Research Laboratory test. The MHA-TP test failed to confirm reactivity in 13 of 79 sera which were reactive in the Venereal Disease Research Laboratory test and in 24 of 101 sera which were reactive in the rapid plasma reagin 18-mm circle card test. Testing another production lot of MHA-TP reagents resulted in even poorer correlation. The reactivity of the MHA-TP test in primary syphilis appeared to vary with the sensitivity of the production lot of reagents.

Fluorescent Antibody Technique↗
PubMed1984

Evaluation of the hemagglutination treponemal test for syphilis.

The sensitivity and specificity of the hemagglutination treponemal test for syphilis (HATTS) was compared with the fluorescent treponemal antibody-absorption test (FTA-ABS) with 491 sera. Medical histories were obtained for 153 patients with seroreactivity in either treponemal test. Overall correlation with patient history was 96.7% for the FTA-ABS and 93.9% for the HATTS. False-negative HATTS occurred in primary, late-latent (greater than 2 years), and treated syphilis. The reproducibility of the HATTS and FTA-ABS is equivalent. HATTS is an acceptable treponemal confirmatory test for syphilis, but the FTA-ABS should be used to resolve diagnostic discrepancies.

Antibodies, Bacterial↗
PubMed1986

Treponemal antibody-absorbent enzyme immunoassay for syphilis.

An enzyme immunoassay for the diagnosis of syphilis (ELISA-SY) was developed with solid-phase extracts of Treponema pallidum, specimen diluent containing Reiter treponeme absorbent, and three 30-min incubations. The ELISA-SY results were determined in comparison with a standardized positive control and reported as a percentage of strong positive control. In tests with 1,005 serum samples from a venereal disease clinic and other sources, 98.2% agreement was found with fluorescent treponemal antibody-absorption (FTA-ABS) results, and 98.3% agreement was found with T. pallidum passive hemagglutination (PHA) findings. Only 1 of 29 sera originally considered to be biologically false-positive by ELISA-SY; the latter specimen was also positive by PHA and FTA-ABS tests performed in our laboratories. Serum samples from clinically diagnosed syphilitics (16 primary-stage isolates, 7 secondary-stage isolates, and 3-latent-stage isolates) were all positive by ELISA-SY, FTA-ABS, and PHA. Serum samples from 51 newborns suspected of having syphilis on the basis of positive cardiolipin flocculation tests showed 98% agreement of ELISA-SY results with FTA-ABS and PHA findings. Sera from all 61 patients with a variety of autoimmune and other diseases known to be associated with biologically false-positive reactions for syphilis were negative by this ELISA-SY. The specificity of the ELISA procedure for T. pallidum antibody was also confirmed immunologically by blocking experiments.

Antibodies↗
PubMed1989

Visuwell Reagin, a non-treponemal enzyme-linked immunosorbent assay for the serodiagnosis of syphilis.

A urease-based enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of reagin antibodies in serum. Visuwell Reagin (ADI Diagnostics Inc., Rexdale, Ontario, Canada) is a non-treponemal screening test for the serodiagnosis of syphilis which has the benefits of large batch testing, automatability, and objective interpretation of results. Unheated, undiluted sera are incubated in 96-well microtiter plates coated with a modified cardiolipin-lecithin-cholesterol antigen. Antibody bound to the plate is detected by an anti-human immunoglobulin G-urease conjugate. The procedure consists of three steps, with a total test time of 60 min. Visuwell Reagin ELISA was compared with the Venereal Disease Research Laboratory (VDRL) test and the reagin screening test (RST) with the following results. For ELISA versus the VDRL test, the sensitivities for untreated syphilis (n = 37) were 97.3% for both ELISA and the VDRL test, the confirmatory positive values (n = 79) were 84.8% for ELISA and 72.2% for the VDRL test, and the specificities for normal samples (n = 1,327) were 98.8% for ELISA and 99.5% for the VDRL test. For ELISA versus RST, the sensitivities for untreated syphilis (n = 57) were 94.7% for ELISA and 87.7% for RST, the confirmatory positive values (n = 26) were 96.2% for ELISA and 92.3% for RST, and the specificities for normal samples (n = 1,891) were 99.6% for ELISA and 99.3% for RST. The overall concordance values of ELISA with VDRL test and RST were 96.7 and 97.9%, respectively. The specificity of ELISA compared with that of RST may be underestimated, since confirmatory data were not available for all apparent false-positive samples. Visuwell Reagin had increased sensitivity and similar specificity compared with flocculation tests.

Antibodies↗
PubMed1990

Evaluation of immunoglobulin M western blot analysis in the diagnosis of congenital syphilis.

Western immunoblots of solubilized Treponema pallidum antigens were reacted with sera and cerebrospinal fluid (CSF) and developed with enzyme-conjugated antibodies to immunoglobulin M (IgM). A blot was considered positive if reactions included bands at the 47-, 17-, and 15.5-kilodalton positions along with a variable pattern at other low-molecular-weight positions. Sera from 23 of 25 symptomatic infants diagnosed with congenital syphilis yielded positive reactions. Of 80 asymptomatic infants considered at risk for developing symptomatic infection, 16 exhibited IgM patterns consistent with those seen in congenital syphilis, although 5 of these 16 gave reactions that were equivocal. To exclude false-positive reactions due to IgM rheumatoid factor, sera were fractionated and the IgM fractions were retested. Only the five initially equivocal sera gave nonreactive blots with the IgM fractions, whereas all others gave more prominent reactions that were qualitatively similar to those seen in serum samples. Sera from 18 normal infants failed to show any IgM reactivity to T. pallidum antigens on Western blots. The IgM Western blot was both more sensitive and more specific than the fluorescent treponemal antibody-absorbed (IgM) test using fractionated serum. Of the 17 CSF samples from infants with symptomatic congenital syphilis, 14 showed IgM reactivity in Western blots, whereas only 12 had a reactive CSF in the Venereal Disease Research Laboratory test. Our results indicate that this technique can be used to identify both symptomatic and asymptomatic infection in infants with T. pallidum, in some cases before standard serologic studies can confirm the diagnosis.

Antibodies, Bacterial↗
PubMed1993

Analysis of the humoral response to the flagellin protein of Borrelia burgdorferi: cloning of regions capable of differentiating Lyme disease from syphilis.

Selected regions of the Borrelia burgdorferi flagellin gene (fla) that exhibit high or low homology with related genes from other bacterial species were amplified by the polymerase chain reaction and expressed as fusion proteins in Escherichia coli. Purified fusion proteins were assayed for antibody reactivity in a microtiter plate enzyme-linked immunosorbent assay with sera from Lyme disease patients as well as syphilitic and normal sera. Immunoglobulin G antibody from Lyme disease patient sera reacted predominantly with the central portion of the protein. The region of the flagellin protein encompassing amino acids 64 to 311 detected nearly all of the immunoglobulin G-positive Lyme sera and only reacted with 1 of 26 syphilis patient serum samples. In contrast, 12 of 26 syphilis patient serum samples and 2 of 47 normal serum samples reacted with the amino terminus of the flagellin protein, whereas 4 of 26 syphilis patient serum samples and 7 of 47 normal serum samples reacted with the carboxyl terminus. The central region containing amino acids 64 to 311 may be employed diagnostically to differentiate antibodies to B. burgdorferi from antibodies to Treponema pallidum. In addition, this region also was recognized by immunoglobulin M in the Lyme patient sera, indicating its potential usefulness as a marker for early Lyme disease.

Antibodies, Bacterial↗
PubMed1978

Counterimmunoelectrophoresis of Reiter Treponeme Axial filaments as a diagnostic test for syphilis.

Purified axial filaments from the cultivable Reiter treponeme, previously shown to share an antigenic component with pathogenic Treponemia pallidum, were evaluated as antigen in a diagnostic test for syphilis. Antibody to the filaments was revealed by counterimmunoelectrophoresis. Conditions that produced optimal results in the test were established. A total of 343 sera from normal individuals, biological false-positive reactors, and from patients in the different stages of syphilis were subjected to the test. The results indicate the test to be sensitive and highly specific for detecting treponemal antibodies in human syphilis.

Antigens, Bacterial↗
PubMed1988

Congenital syphilis in the newborn.

We studied 53 newborn babies with congenital syphilis. The common clinical features seen were low birth weight, hepatosplenomegaly, anaemia, jaundice, and symmetrical superficial desquamation of the skin affecting palms and soles. The presence of these clinical signs is highly suggestive of early congenital syphilis. Hydrops fetalis without rhesus or ABO isoimmunisation should always arouse the suspicion of congenital syphilis.

Anemia, Neonatal↗
PubMed2004

HIV, syphilis infection, and sexual practices among transgenders, male sex workers, and other men who have sex with men in Jakarta, Indonesia.

OBJECTIVES: To establish the prevalence of HIV, syphilis, and sexual risk behaviour among three groups of men who have sex with men in Jakarta, Indonesia, and to investigate sexual links between these men and broader heterosexual populations. METHODS: Anonymous, cross sectional surveys among community recruited transgender and male sex workers and self recognised men who have sex with men (MSM) were undertaken in mid-2002 in Jakarta, Indonesia. Places where transgender and male sex workers sell sex and where men go to meet non-commercial male sex partners were mapped. Probability samples were selected for the sex worker populations, while a mixed probability and convenience sample was drawn for self recognised MSM. Blood was drawn for HIV and syphilis serology and community interviewers administered a standardised questionnaire. RESULTS: HIV prevalence was 22% among transgender sex workers, 3.6% among male sex workers, and 2.5% among self recognised MSM, and syphilis prevalence was 19.3%, 2.0% and 1.1% respectively. 59.3% of transgender sex workers and 64.8% of male sex workers reported recent unprotected anal intercourse with clients, and 53.1% of other MSM reported unprotected anal sex with male partners. Some 54.4% of male sex workers and 18.3% of other MSM reported female partners in the preceding year. CONCLUSION: HIV has reached substantial levels among transgender sex workers, and is not negligible in other MSM groups. Risk behaviour is high in all subpopulations, and bisexual behaviour is common, meaning the threat of a wider epidemic is substantial. Prevention programmes targeting male-male sex are needed to reduce this threat.

Adult↗
PubMed2006

Male circumcision and risk of syphilis, chancroid, and genital herpes: a systematic review and meta-analysis.

OBJECTIVES: Male circumcision is associated with reduced risk of HIV infection. This may be partly because of a protective effect of circumcision on other sexually transmitted infections (STI), especially those causing genital ulcers, but evidence for such protection is unclear. Our objective was to conduct a systematic review and meta-analyses of the associations between male circumcision and infection with herpes simplex virus type 2 (HSV-2), Treponema pallidum, or Haemophilus ducreyi. METHODS: Electronic databases (1950-2004) were searched using keywords and text terms for herpes simplex, syphilis, chancroid, ulcerative sexually transmitted diseases, or their causative agents, in conjunction with terms to identify epidemiological studies. References of key articles were hand searched, and data were extracted using standardised forms. Random effects models were used to summarise relative risk (RR) where appropriate. RESULTS: 26 articles met the inclusion criteria. Most syphilis studies reported a substantially reduced risk among circumcised men (summary RR = 0.67, 95% confidence interval (CI) 0.54 to 0.83), although there was significant between study heterogeneity (p = 0.01). The reduced risk of HSV-2 infection was of borderline statistical significance (summary RR = 0.88, 95% CI 0.77 to 1.01). Circumcised men were at lower risk of chancroid in six of seven studies (individual study RRs: 0.12 to 1.11). CONCLUSIONS: This first systematic review of male circumcision and ulcerative STI strongly indicates that circumcised men are at lower risk of chancroid and syphilis. There is less association with HSV-2. Potential male circumcision interventions to reduce HIV in high risk populations may provide additional benefit by protecting against other STI.

Chancroid↗
PubMed1975

Application of the enzyme-linked immunosorbent assay (ELISA) in the serodiagnosis of syphilis.

The enzyme-linked immunosorbent assay (ELISA) technique, using an ultrasonicate of Treponema pallidum as antigen, has been evaluated as a serological test for syphilis. It is concluded that the test is simple, reliable, and relatively quick and that its sensitivity in all stages of syphilis is equal to the FTAABS test. Because its specificity is probably also high, ELISA might be used in future as a first-line screening test for the serodiagnosis of syphilis.

Adsorption↗