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Detecting anti-Treponema hyodysenteriae antibodies in swine serum using immunofluorometry.

An indirect fluorescent antibody test was adapted for measuring serum anti-Treponema hyodysenteriae antibodies with a fluorometer. The immunofluorescence was recorded as fluorescent signal units. Cultures of T. hyodysenteriae and Treponema innocens were used as antigen. There was a significant (P less than 0.01) correlation between the immunofluorescence recorded with the fluorometer and that evaluated visually with a microscope. The swine exposed orally to swine dysentery infective inoculum and subsequently hyperimmunized by the intravenous inoculation of live cultures of T. hyodysenteriae had the highest average fluorescent signal unit, which was 104.5. There was a significant (P less than 0.01) correlation between the level of anti-T. hyodysenteriae antibody and the interval length between the last day of diarrhea and the day of bleeding. However, in measuring fluorescent signal units in serum from swine infected with nonpathogenic large spirochetes, (T. innocens), there was also a significant (P less than 0.01) correlation between T. hyodysenteriae and T. innocens as antigen. The coefficient of variation of the average fluorescent signal unit for a highly positive serum and a highly negative serum between 16 runs of assays were 5.7% and 19% respectively; the coefficient of variation of the average fluorescent signal unit for duplicate samples on 358 serum samples tested was 5.8%.

Animals↗

Scintigraphic diagnosis of syphilitic lesions in rabbits by radiolabelled monoclonal antibodies specific for Treponema pallidum.

Immunoscintigraphy with radiolabelled monoclonal antibodies has been widely used to detect solid tumours. The purpose of this study was to investigate its potential for the specific localization of syphilitic lesions. F(ab')2 fragments were prepared from murine monoclonal antibodies against Treponema pallidum produced in our laboratory and labelled with 131I. Bilateral testicular infections were created in rabbits by inoculation with T. pallidum or Staphylococcus aureus. Seven to 10 days after inoculation, radiolabelled antibodies were injected intravenously. Serial gamma images were then taken 2 h after the injection and at 24 h intervals thereafter. Beginning as early as 2 h post-injection, the testicles could be visualized with either specific or non-specific antibodies. Gamma images in the monoclonal antibody-treated, T. pallidum-infected group persisted up to 48 h post-injection. Lesions were not discernible from background in the S. aureus-infected group injected with the monoclonal antibody and the S. aureus-infected and T. pallidum-infected groups injected with the polyclonal antibody at 24 h post-injection or later. Therefore, due to its ability to differentiate between specific and non-specific antibody-generated images from 24 h post-injection, immunoscintigraphy using monoclonal antibodies specific for T. pallidum may be employed as one of the methods to diagnose difficult cases of syphilitic internal organ involvement as well as syphilis infection in seronegative HIV-infected patients.

Animals↗

Detection of treponemal DNA in the CSF of patients with syphilis and HIV infection using the polymerase chain reaction.

The polymerase chain reaction (PCR) was used to detect Treponema pallidum DNA in the cerebrospinal fluid (CSF) of patients with and without syphilis. The CSF from 10 of 19 patients with positive serological tests for syphilis who were being investigated for late syphilis were treponemal DNA-positive. In contrast, the CSF from only one of 30 patients with no known history of syphilis was DNA-positive. CSF from 28 HIV-positive patients was also tested. Fourteen of these patients had central nervous system (CNS) disease and seven were DNA-positive, whereas none of the 14 without CNS disease were DNA-positive. Five of the seven DNA-positive patients had a history of syphilis. Such a history in an HIV-positive patient who had CNS disease was predictive of finding treponemal DNA in the CSF. The PCR had a sensitivity of 47% and a specificity of 93% for detecting a known history of syphilis and is a potentially useful tool in treponemal diagnosis.

Acquired Immunodeficiency Syndrome↗

Surface-associated antigens of Treponema pallidum concealed by an inert outer layer.

Soluble antigens of Treponema pallidum were examined by two dimensional immunoelectrophoresis against antisera from infected or artificially immunized rabbits. Concentrated suspensions of intact cells did not release antigens after storage at 4 degrees, incubation at 37 degrees, or vortex mixing. Antigens were released after disintegration of treponemes by ultrasonic vibration, or by treatment with non-ionic or anionic detergents. An antigenic component of sonicated treponemes, present in both the non-pathogenic, cultivable Reiter treponeme and T. Pallidum, was identified as axial filament. The combination of antibody with unfixed whole organisms was monitored by an indirect fluorescent antibody method, and whereas antibody did not combine with intact organisms, detergent-treated organisms were highly reactive. Immune electron microscopy showed that whereas in intact treponemes, axial filaments were unable to combine with antibody, detergent treatment allowed access to axial filaments by antibody. In intact treponemes the axial filaments are thought to be located beneath the outer membrane, which may thus comprise the postulated antigenically inert outer layer.

Animals↗

Evaluation of new anti-infective drugs for the treatment of syphilis. Infectious Diseases Society of America and the Food and Drug Administration.

Syphilis is caused by Treponema pallidum, a spirochetal bacterium pathogenic only for humans. The clinical course of disease is divided into three stages interspersed by periods of latency. Penicillin remains the treatment of choice for all stages of infection; tetracycline or erythromycin may be used as therapeutic alternatives in defined circumstances. Patients enrolled in clinical trials should be evaluated clinically, microscopically, and serologically for the presence of the spirochete. All participants, after undergoing counseling and giving informed consent, should be tested for infection with human immunodeficiency virus. Specific criteria exist for diagnosis of syphilis and response to therapy. It may be desirable to perform a small, uncontrolled, open trial of a new anti-infective drug for the collection of preliminary evidence of efficacy. A larger-scale, randomized, active-control comparative clinical trial is necessary to prove efficacy.

Animals↗

Tpr homologs in Treponema paraluiscuniculi Cuniculi A strain.

Treponema paraluiscuniculi, the etiologic agent of rabbit venereal syphilis, is morphologically indistinguishable from Treponema pallidum subsp. pallidum (T. pallidum), the human syphilis treponeme, and induces similar immune responses and histopathologic changes in the infected host. Because of their high degree of relatedness, comparative studies are likely to identify genetic determinants that contribute to pathogenesis or virulence in human syphilis. The tpr (Treponema pallidum repeat) genes are believed to code for potential virulence factors. In this study, we identified 10 tpr homologs in Treponema paraluiscuniculi Cuniculi A strain and determined their sequence architecture. Half of this group of paralogous genes were predicted to be nonfunctional due to the presence of frameshifts and premature stop codons. Furthermore, the immune response against the T. paraluiscuniculi Tpr homologs in long-term-infected rabbits was studied by enzyme-linked immunosorbent assay and lymphocyte proliferation assay, showing that TprK is the only target of the antibody and T-cell responses during experimental infection and emphasizing the importance of this putative virulence factor in venereal treponematosis.

Amino Acid Sequence↗

Oral and dental infections with anaerobic bacteria: clinical features, predominant pathogens, and treatment.

Microbial populations colonizing the teeth are a major source of pathogens responsible for oral and dental infections, including periodontal diseases, gingivitis, pericoronitis, endodontitis, peri-implantitis, and postextraction infections. Each entity has distinct clinical and microbial features. Bacterial species associated with oral infections include Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus, Campylobacter rectus, Eubacterium species, Fusobacterium nucleatum, Eikenella corrodens, and Peptostreptococcus micros. Treponema pallidum-related spirochetes have been associated with acute necrotizing ulcerative gingivitis. Porphyromonas endodontalis appears to be specifically related to endodontic infections. Oral infections in medically compromised patients, including those with AIDS, are associated with similar species and are usually complicated by superinfection with enteric and Candida species. Isolation of species causing oral infections requires the collection of appropriate samples and the use of strictly anaerobic techniques. Rapid selective culture, immunofluorescence, and DNA probe methods have been developed for the identification of these oral species. The varied measures required in the management of oral and dental infections may include antimicrobial therapy. Accurate microbiological diagnosis, including antibiotic susceptibility testing, is indicated for cases that do not respond to therapy.

Bacteria, Anaerobic↗

Experimental syphilis and serological examination for treponematosis in hares.

Of 202 captive hares studied, many of which had lesions on their external genital organs or testicular atrophy or both, 27% had positive serological tests for syphilis although dark-field examination of extracts of atrophic testes was negative. A total of 12 hares that were nonreactive for the serological test for syphilis was inoculated with Treponema pallidum, 9 intratesticularly and 3 intradermally. Six of the animals inoculated intratesticularly exhibited orchitis after 7 days with an associated accumulation of treponemes. No chancres developed in the intracutaneously inoculated animals during a 27-day period of observation. These results provide additional evidence to support the contention that endemic treponematosis occurs in wild hares and suggest that hares are moderately resistant to experimental infection with T. pallidum.

Animals↗

Identification of novel virulence-associated genes via genome analysis of hypothetical genes.

The sequencing of bacterial genomes has opened new perspectives for identification of targets for treatment of infectious diseases. We have identified a set of novel virulence-associated genes (vag genes) by comparing the genome sequences of six human pathogens that are known to cause persistent or chronic infections in humans: Yersinia pestis, Neisseria gonorrhoeae, Helicobacter pylori, Borrelia burgdorferi, Streptococcus pneumoniae, and Treponema pallidum. This comparison was limited to genes annotated as hypothetical in the T. pallidum genome project. Seventeen genes with unknown functions were found to be conserved among these pathogens. Insertional inactivation of 14 of these genes generated nine mutants that were attenuated for virulence in a mouse infection model. Out of these nine genes, five were found to be specifically associated with virulence in mice as demonstrated by infection with Yersinia pseudotuberculosis in-frame deletion mutants. In addition, these five vag genes were essential only in vivo, since all the mutants were able to grow in vitro. These genes are broadly conserved among bacteria. Therefore, we propose that the corresponding vag gene products may constitute novel targets for antimicrobial therapy and that some vag mutants could serve as carrier strains for live vaccines.

Animals↗

Risk factors for HTLV-I among heterosexual STD clinic attenders.

Human T-cell lymphotropic virus type 1 (HTLV-I) status was assessed in 994 patients attending a sexually transmitted disease (STD) clinic in Kingston, Jamaica, between November 1990 and January 1991 for a new STD complaint. Of 515 heterosexual men, 36 (7.0%) were HTLV-I seropositive, as were 38 (7.9%) of 479 women. HTLV-I seroprevalence increased with age in women. A history of blood transfusion was associated with HTLV-I in both sexes, significantly so in men [odds ratio (OR) 4.7, confidence interval (CI) 1.1-17 for men; OR 1.9, CI 0.6-5.0 for women]. Further analysis excluded all persons reporting a transfusion. On multiple logistic regression analysis, independent associations with HTLV-I infection in men were shown for marital status (OR 3.5, CI 1.2-10 for married/common law vs. single/visiting unions), agricultural occupation (OR 9.0, CI 2.0-41), bruising during sex (OR 2.9, CI 1.0-8.1), > or = 15 years at first sexual intercourse (OR 2.9, CI 1.0-8.2), and a positive test for hepatitis B surface antigen (OR 7.3, CI 1.2-52). In women, associations were shown for two or more sex partners in the 4 weeks prior to complaint (OR 4.9, CI 1.8-13), 11 or more lifetime sexual partners (OR 5.9, CI 1.3-27), aged < 15 years at first sexual intercourse (OR 2.3, 1.0-5.4), bruising during sex (OR 2.7, CI 1.1-6.6), microhaemagglutination-Treponema pallidum positivity (OR 3.6, CI 1.6-8.4), and human immunodeficiency virus infection (OR 14, CI 2.1-92).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Porphyromonas gingivalis vesicles enhance attachment, and the leucine-rich repeat BspA protein is required for invasion of epithelial cells by "Tannerella forsythia".

The human oral cavity harbors more than 500 species of bacteria. Periodontitis, a bacterially induced inflammatory disease that leads to tooth loss, is believed to result from infection by a select group of gram-negative periodontopathogens that includes Porphyromonas gingivalis, Treponema denticola, and "Tannerella forsythia" (opinion on name change from Tannerella forsythensis pending; formerly Bacteroides forsythus). Epithelial cell invasion by periodontopathogens is considered to be an important virulence mechanism for evasion of the host defense responses. Further, the epithelial cells with invading bacteria also serve as reservoirs important in recurrent infections. The present study was therefore undertaken to address the epithelial cell adherence and invasion properties of T. forsythia and the role of the cell surface-associated protein BspA in these processes. Further, we were interested in determining if P. gingivalis, one of the pathogens frequently found associated in disease, or its outer membrane vesicles (OMVs) could modulate the epithelial cell adherence and invasion abilities of T. forsythia. Here we show that epithelial cell attachment and invasion by T. forsythia are dependent on the BspA protein. In addition, P. gingivalis or its OMVs enhance the attachment and invasion of T. forsythia to epithelial cells. Thus, interactions between these two bacteria may play important roles in virulence by promoting host cell attachment and invasion.

Antigens, Bacterial↗

Development of a real-time PCR assay to detect Treponema pallidum in clinical specimens and assessment of the assay's performance by comparison with serological testing.

The incidence of infectious syphilis in men who have sex with men and human immunodeficiency virus-infected patients has increased steadily in Victoria, Australia, since 2002. A TaqMan real-time PCR assay targeting the polA gene of Treponema pallidum (TpPCR) was developed. The analytical sensitivity of the assay was estimated to be 1.75 target copies per reaction. Initially, the assay was used to test a variety of specimens (excluding blood) from 598 patients. Of the 660 tests performed, positive PCR results were obtained for 55 patients. TpPCR results were compared with serology results for 301 patients being investigated for early syphilis. Of these patients, 41 were positive by both TpPCR and serology, 246 were negative by both TpPCR and serology, 4 were TpPCR positive but negative by serology, and 10 were TpPCR negative but showed evidence of recent or active infection by serology. Directly compared with serology, TpPCR showed 95% agreement, with a sensitivity of 80.39% and a specificity of 98.40%. Potential factors leading to the discrepant results are discussed. Concurrent serology on 21 patients with TpPCR-positive primary syphilitic lesions demonstrated that a panel of current syphilis serological tests has high sensitivity for the detection of early syphilis. We found that TpPCR is a useful addition to serology for the diagnosis of infectious syphilis. Direct comparison with other T. pallidum PCR assays will be required to fully assess the limitations of the assay.

Bacterial Proteins↗

The Use of Molecular Techniques for the Diagnosis and Epidemiologic Study of Sexually Transmitted Infections.

Molecular diagnostic tests are more sensitive and, in many cases, more specific than conventional laboratory methods for the detection of sexually transmitted infections. Here, we review recently developed molecular methods for the diagnosis and subtyping of the most common sexually transmitted infections: infections caused by Chlamydia trachomatis, Neisseria gonorrhoeae, human papillomavirus, Trichomonas vaginalis, and the agents of genital ulcer disease (Haemophilus ducreyi, herpes simplex virus, Treponema pallidum, and Calymmatobacterium granulomatis). We also provide an overview of the laboratory diagnostic tests and clinical specimens to use when infection with these agents is suspected.

Journal Article↗

Low HIV and high STD among commercial sex workers in a brothel in Bangladesh: scope for prevention of larger epidemic.

The present study documents the first systematic assessment of a brothel in Bangladesh in terms of sexually transmitted disease (STD) and human immunodeficiency virus (HIV). A cross sectional study was undertaken on brothel-based commercial sex workers (CSWs) selected systematic random sampling to assess the prevalence of STDs and HIV among CSWs in a brothel setting. Two hundred and ninety-six CSWs were selected from a brothel with a population of 593 women. Following informed consent, endocervical and blood samples were obtained for the diagnosis of genital chlamydia, gonorrhoea, HIV and syphilis respectively. In addition, another 170 consecutive blood samples were collected from the total CSW population for HIV tests. All blood samples for HIV testing were made anonymous by removing patient identifiers before testing. Endocervical specimens were tested by polymerase chain reaction (PCR) for the diagnosis of genital chlamydia and gonorrhoea. Syphilis and HIV infections were diagnosed by serology. One hundred and sixty-nine (57.1%) of the women were Treponema pallidum haemagglutination (TPHA)-positive, 20 (6.8%) of the women were Venereal Disease Research Laboratory (VDRL)-positive at a greater than 1:8 dilution. Eighty-two (28%) of the women were found to be infected either by gonorrhoea or chlamydia. No HIV antibody was found in any of the 466 blood samples. A high prevalence of STDs and low prevalence of HIV in the CSWs in Bangladesh suggest potential for the rapid spread of HIV once it is introduced in this high-risk population. The opportunity to control STD and HIV infection in this population should not be missed, in order to prevent a large epidemic in the future.

Bangladesh↗

Macrophage and lymphocyte functions are down-regulated in newborn rabbits.

We have developed an animal model for congenital syphilis. Treponema pallidum is injected intravenously into pregnant rabbits and fetuses are infected in utero. As a prelude to characterizing the immunologic consequences of fetal infection, it was necessary to expand on the baseline information about newborn rabbit immune capabilities. Studies were undertaken to determine splenic macrophage and T lymphocyte functions with emphasis on newer immunologic parameters. Newborns aged 2 weeks were compared to adults. Macrophage capabilities in newborn rabbits differed from those of their adult counterparts. These cells produced similar basal levels of interleukin 1 (IL-1) but failed to respond to the IL-1 stimulants of lipopolysaccharide (LPS) or T. pallidum. Macrophages also exhibited diminished levels of la expression and increased levels of prostaglandin E2 (PGE2) secretion. T lymphocyte functions were altered in newborn spleen preparations. Following concanavalin A (Con A) stimulation, interferon gamma production was half that of adults; in direct contrast, IL-2 production was twice that of adults. Con A-induced lymphocyte proliferation was markedly decreased in newborn preparations. This diminished response resulted from down-regulation rather than immaturity. When newborn splenic cells were stimulated with Con A in the presence of indomethacin, anti-transforming growth factor (anti-TGF), or exogenous IL-1/IL-2, better proliferation resulted. PGE2, which is well established as a down-regulator of newborn immune functions in human and mouse systems, also appears to play a role in suppressing newborn rabbit functions. TGF is a potent suppressor of a number of adult immunologic reactions. This is the first documentation of the potential role of this factor in down-regulating newborn immune capabilities. These findings provide a framework for future investigations of our congenital syphilis model.

Animals↗

Sexually transmitted diseases and sexual behaviour as risk factors for HIV-2 infection in Bissau, Guinea Bissau.

Patients enrolled in a hospital-based case control study (n = 1009) and a follow-up study (n = 130) of HIV-2 infection in Bissau were investigated with regard to sexual behaviour and sexually transmitted diseases (STD). A history of genital ulcer was more frequent in HIV-2 infected male patients (46.4%, 13 out of 28) than in seronegative male patients (17.9%, 7 out of 39) (P = 0.05). Serological evidence of a previous syphilitic infection was significantly related to HIV-2 infection (23%, 12 out of 52, of HIV-2 seropositive patients had a positive Treponema pallidum haemagglutination test compared with 7.1%, 5 out of 70, of seronegative patients, P = 0.025) and was equally frequent in men and women. There was no significant difference in number of non-marital sexual partners and frequency of intercourse between HIV-2 seropositive and seronegative individuals. The seroprevalence of HIV-2 infection was significantly higher among patients with a monogamous marriage, 23.1% (59 out of 255) among males and 31.3% (64 out of 204) among females, than among patients with an officially polygamous marriage, 10.3% (8 out of 77) among males and 11.8% (7 out of 59) among females (P = 0.025 for males and 0.005 for females). Infection-control measures directed at patients with STD should be promoted in Guinea-Bissau. Further research on sexual behaviour and attitudes is needed.

Acquired Immunodeficiency Syndrome↗

Markers of past syphilis in HIV infection comparing Captia Syphilis G anti-treponemal IgG enzyme immunoassay with other treponemal antigen tests.

The sensitivity of the Captia Syphilis G anti-treponemal IgG enzyme immunoassay (EIA-IgG) was compared with the Treponema pallidum haemagglutination assay (TPHA) and the Fluorescent Treponemal Antibody Absorbed (FTA-Abs) test as a marker for past syphilis in 28 HIV-infected and 31 HIV-negative patients with a past history of syphilis. The specificity of EIA-IgG was compared in 89 patients without a history of syphilis who were known to be HIV antibody positive with a control group of 89 patients who had tested HIV negative. In patients with a past history of syphilis each treponemal test (EIA-IgG, TPHA and FTA-Abs) gave a lower sensitivity (82%, 86%, 79%) in the HIV-positive group than in the HIV-negative group (97%) but the difference was significant only in the case of the FTA-Abs test (P < 0.05). In the HIV-positive patients 11% (3/28) were negative in all 3 treponemal tests while 25% (7/28) were negative in at least one treponemal test. In patients without a past history of syphilis the EIA-IgG antibody index in the HIV-positive group (0.436) was significantly higher than in the HIV-negative group (0.378): the specificity, however, was similar in the HIV-positive (100%) and HIV-negative group (99%). We conclude that the Captia Syphilis G anti-treponemal IgG enzyme immunoassay is of similar specificity in HIV-positive and HIV-negative patients and is of similar sensitivity to the TPHA and FTA-Abs as a marker of past syphilis in HIV-infected patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Bacterial↗