Different Stages Of Syphilis Research Articles

Serological Diagnosis of Syphilis: Enzyme‐Linked Immunosorbent Assay to Measure Antibodies to Individual Recombinant Treponema pallidum Antigens

We standardized an indirect ELISA for measurement of serum antibody levels to four individual treponemal recombinant proteins that have been commonly used in a number of commercial EIAs, mostly as a mixture of antigens. When tested with 127 syphilis‐negative and 37 secondary syphilis sera, ELISA O.D.s obtained for each of the four antigens clearly distinguished between these two groups of samples. Sensitivity and specificity of 100% was obtained with the current set of samples. Further evaluations with sera from different stages of syphilis can help to define the applications of this ELISA test for each of the four antigens studied.

Syphilis depicted by the Greek moulages: a picture of skin manifestations in former times

A great part of the medical wax moulages produced in the beginning of the last century depicts the various clinical signs of syphilis. Based on the study of a large Greek moulage collection, the different stages of syphilis are described. Moulages may still have the original scope as teaching objects, presenting clinical manifestations seldom seen to day and thus more likely of being overseen by the clinicians.

Syphilis does not seem to involve virological and immunological course of concurrent HIV disease

Syphilis does not seem to involve virological and immunological course of concurrent HIV disease

Western immunoblotting with five Treponema pallidum recombinant antigens for serologic diagnosis of syphilis.

Five immunodominant Treponema pallidum recombinant polypeptides (rTpN47, rTmpA, rTpN37, rTpN17, and rTpN15) were blotted onto strips, and 450 sera (200 from blood donors, 200 from syphilis patients, and 50 potentially cross-reactive) were tested to evaluate the diagnostic performance of recombinant Western blotting (recWB) in comparison with in-house whole-cell lysate antigen-based immunoblotting (wclWB) and T. pallidum hemagglutination (MHA-TP) for the laboratory diagnosis of syphilis. None of the serum specimens from blood donors or from potential cross-reactors gave a positive result when evaluated by recWB, wclWB, or MHA-TP. The evaluation of the immunoglobulin G immune response by recWB in sera from patients with different stages of syphilis showed that rTmpA was the most frequently identified antigen (95%), whereas only 41% of the specimens were reactive to rTpN37. The remaining recombinant polypeptides were recognized as follows: rTpN47, 92.5%; rTpN17, 89.5%; and rTpN15, 67.5%. The agreement between recWB and MHA-TP was 95.0% (100% with sera from patients with latent and late disease), and the concordance between wclWB and MHA-TP was 92.0%. The overall concordance between recWB and wclWB was 97.5% (100% with sera from patients with secondary and late syphilis and 94.6 and 98.6% with sera from patients with primary and latent syphilis, respectively). The overall sensitivity of recWB was 98.8% and the specificity was 97.1% with MHA-TP as the reference method. These values for sensitivity and specificity were slightly superior to those calculated for wclWB (sensitivity, 97.1%, and specificity, 96.1%). With wclWB as the standard test, the sensitivity and specificity of recWB were 98.9 and 99.3%, respectively. These findings suggest that the five recombinant polypeptides used in this study could be used as substitutes for the whole-cell lysate T. pallidum antigens and that this newly developed recWB test is a good, easy-to-use confirmatory method for the detection of syphilis antibodies in serum.

The ability of peripheral blood mononuclear cells (PBMC) of syphilitic patients to produce IL-2

The cell-mediated immune response of importance in protection against Treponema pallidum, is distinctly suppressed in some stages of the disease. This may be a result of decreased ability of cells to produce IL-2, or IL-2 absorption by different factors. The experiments were designed to evaluate the ability of peripheral blood mononuclear cells (PBMC) of patients with different stages of syphilis to produce IL-2, and to investigate the causes which could possibly limit its activity. The ability of the PBMC of syphilitic patients to produce IL-2 develops at the beginning of the disease, reaching a maximum in primary seropositive syphilis. In the next stages of the disease this capability is distinctly lowered. The lowest was in malignant syphilis and tabes dorsalis, i.e. during severe disease. Absorption of adherent cells from PBMC increased the ability of lymphocytes to produce IL-2. The highest level of this interleukin was observed at the stages of the disease where suppression was the deepest. Sera of both control and syphilitic patients contained IL-2 inhibitor. Its level was the highest in early and late latent syphilis where no symptoms of disease were present. In all syphilitic sera a distinctly elevated level of soluble IL-2 receptors (sIL-2R) was also found. Its high level was noted in sera of patients in which PBMC had the weakest ability to produce IL-2. These findings suggest that sIL-2R may be bound to IL-2 and in this way would lead to weakening of T cell function and of resistance against Treponema pallidum infection.

The ability of peripheral blood mononuclear cells (PBMC) of syphilitic patients to produce IL-2

The cell-mediated immune response of importance in protection against Treponema pallidum, is distinctly suppressed in some stages of the disease. This may be a result of decreased ability of cells to produce IL-2, or IL-2 absorption by different factors. The experiments were designed to evaluate the ability of peripheral blood mononuclear cells (PBMC) of patients with different stages of syphilis to produce IL-2, and to investigate the causes which could possibly limit its activity. The ability of the PBMC of syphilitic patients to produce IL-2 develops at the beginning of the disease, reaching a maximum in primary seropositive syphilis. In the next stages of the disease this capability is distinctly lowered. The lowest was in malignant syphilis and tabes dorsalis, i.e. during severe disease. Absorption of adherent cells from PBMC increased the ability of lymphocytes to produce IL-2. The highest level of this interleukin was observed at the stages of the disease where suppression was the deepest. Sera of both control and syphilitic patients contained IL-2 inhibitor. Its level was the highest in early and late latent syphilis where no symptoms of disease were present. In all syphilitic sera a distinctly elevated level of soluble IL-2 receptors (sIL-2R) was also found. Its high level was noted in sera of patients in which PBMC had the weakest ability to produce IL-2. These findings suggest that sIL-2R may be bound to IL-2 and in this way would lead to weakening of T cell function and of resistance against Treponema pallidum infection.

Pathogen-Specific Humoral Response in Treponema pallidum-Infected Humans, Rabbits, and Guinea Pigs

Molecular analysis of the humoral response to pathogen-specific polypeptides was done using sera from patients at different stages of syphilis and sera from Treponema pallidum-infected rabbits and guinea pigs collected at various times after infection. The sera were examined by ELISA, fluorescent treponemal antibody absorption test (FTA-ABS), and immunoblot before and after sequential adsorption with cyanogen bromide-activated Sepharose 4B coupled individually to five sonicated nonpathogenic treponemes. Guinea pig antisera were also adsorbed with rabbit proteins. After adsorption, sera from all three species reacted neither by ELISA nor by immunoblot with nonpathogenic treponemes, nor did guinea pig sera react with rabbit proteins. Regardless of the species of treponema used for initial adsorption, none by itself could effectively remove all cross-reactivities. Nine pathogen-specific polypeptides (15, 17, 33, 37, 39, 43, 45, 47, 97 kDa), eight (15-47 kDa) known as integral membrane or endoflagellar components, were commonly recognized by the tree species when chancre immunity developed. The substantial reduction in immune reactivity after adsorption suggests that most of these peptides consist of specific and common treponemal epitopes. Throughout the various stages of the human disease, pathogen-specific antibodies were exclusively of the IgG isotype.

Factors that inhibit adherence of Treponema pallidum (Nichols strain) to a human fibroblastic cell line: development in serum of patients with syphilis.

Serum samples from 25 patients at five different stages of syphilis were investigated for their ability to inhibit the adherence of pathogenic Nichols treponemes to cultured human fibroblasts. Serum taken from patients at the end of the primary stage showed an appreciable inhibition of treponemal adherence, and maximum inhibition of adherence was produced by serum from patients with secondary syphilis. Some freshly harvested treponemal suspensions were resistant to the adherence inhibition factors in serum from patients with syphilis; after incubation in vitro for 24 hours this resistance was lost. In vitro incubation almost doubled the number of adherent treponemes/fibroblast. These phenomena are discussed in terms of loss and reconstruction of the treponemal outer envelope. This leads to the suggestion that adherence occurs not only at the tips of the treponemes, but that surface components are also implicated, possibly as an initial contact mechanism. The composition of the outer envelope may in this way determine localisation versus dissemination of the treponemes.

Isolation of a Treponema pallidum gene encoding immunodominant outer envelope protein P6, which reacts with sera from patients at different stages of syphilis.

A phage directing the synthesis of an abundant 45-kD Treponema pallidum surface protein was isolated from an EMBL-4 bacteriophage lambda library of T. pallidum DNA. The recombinant phage was identified using an mAb that was directed toward an immunodominant, outer envelope T. pallidum protein designated P6. The recombinant P6 protein possessed the same mol mass as the native treponemal antigen detected from total T. pallidum protein preparations, confirming the cloning of the structural gene for this molecule. Furthermore, E. coli was transformed by a 4.5-kb Eco RI lambda insert fragment subcloned into the plasmid vector pUC19. These transformed cells expressed and translocated the 45-kD protein to their outer membranes. Finally, all sera from patients with different stages of syphilis (primary, secondary, and latent) contained antibody reactive to this protein.

Humoral immune response in human syphilis to polypeptides of Treponema pallidum.

A molecular characterization of the polypeptide antigens of Treponema pallidum reactive with sera from patients with different stages of syphilis is described. Polypeptides of motile, virulent T. pallidum, purified from rabbit testes, were separated on SDS polyacrylamide gels and electrophoretically transferred to nitrocellulose for antigenic analysis ("Western blotting"). Serum IgG from uninfected individuals reacts weakly with three polypeptides of 45,000, 33,000, and 30,000 m.w. In this study patients with primary syphilis have IgM antibody, and all patients with syphilis have IgG antibody to at least four polypeptides of 45,000, 33,000, 30,000, and 15,500 m.w. Antibody to polypeptides of 42,000 and 16,500 m.w. appear to be markers for nonprimary syphilis. These six polypeptides have been termed the major antigenic proteins (MAP) of T. pallidum. Those patients studied with secondary and early latent syphilis acquire antibody to a set of 16 additional polypeptide antigens. Those patients studied with late latent or late syphilis have antibody to a much smaller set of five or four antigens, respectively, in addition to MAP. The results suggest that a correlation exists between acquisition of antibody and the development of "chancre immunity." Additionally, the loss of antibody that characterizes late latent and late syphilis may be associated with the potential development of destructive late syphilis.

Laboratory Evidence for Impaired Cellular Immunity in Different Stages of Syphilis

Abundant evidence suggests that Treponema pallidum (T.p.) escapes humoral immune defence despite the host produces antibodies early in the infection. Since the serologic responses in syphilis have been studied in detail this paper focuses on the cellular immune mechanisms. For this purpose the leukocyte migration inhibition was investigated in 17 patients in different stages of syphilis. Leukocyte migration inhibition assay was performed before, and 7 days, 3 weeks, 2 mo. and 1 yr after start of treatment. Ultrasonicated T.p. were used as antigen corresponding to 5 X 10(6) to 2 X 10(7) Treponema pallida per ml. Controls without antigen, with addition of Concanavalin. A instead of T.p. and using cells of normal volunteers were run. There was no leukocyte migration inhibition before treatment, suggesting nonexistent or depressed cellular immunity in the untreated syphilitic patient. Significant leukocyte migration inhibition, however, was observed as early as 2 days after start of treatment, being most pronounced after 1 week. Hypothetical circulating blocking factors for cellular immune reactions might be present in the untreated syphilitic patient, which become abolished after therapy. Since stimulation with Con A of syphilitic leukocytes gave normal results even before treatment in the syphilitic patient, there might be a specific block of leukocyte migration inhibition against T.p.

Immunoregulatory properties of serum from patients with different stages of syphilis.

The response of lymphocytes from 17 patients with primary, secondary, and tertiary syphilis to phytohaemagglutinin (PHA) and to allogeneic lymphocytes was normal in heterologous serum; however, the responsiveness of cells from some patients with primary and secondary disease was significantly reduced in the presence of autologous serum. As cells from healthy controls invariably responded better to these stimuli in autologous serum, the sera from 81 patients with syphilis were screened for immunosuppressive properties. Sera from 25 primary, 32 secondary, two tertiary, six congenital, and 16 latent cases of syphilis were examined for their ability to reduce the responsiveness of normal cells to PHA. These experiments were performed with test sera as the sole source of serum for the cultures or with test sera added to cultures containing optimally supportive amounts of pooled human plasma.Stimulation of normal cells from one control in human plasma and 20% test serum showed that only in sera from congenital cases of syphilis was the mean response significantly different from the response seen in control sera; a significant increase in the response to stimulation occurred. The range of response to PHA with sera from cases of primary and secondary syphilis was wider than with normal sera. Sera from five (20%) cases of primary and 14 (44%) cases of secondary syphilis appeared to be immunosuppressive. When retested on another sample of normal cells, these sera were consistently immunosuppressive even in the presence of 15% pooled human plasma. Thus, in early syphilis antigenic stimulation may result in the release from suppressor cells of non-specific immunoregulators of cell-mediated immunity. Such phenomena may be a prelude to the development of tolerance to treponemal antigens. In congenital syphilis the development of suppressor cells may be impaired, resulting in the apparent immunostimulatory properties of serum from such cases.

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.

Auto-Immune Phenomena in Syphilitic Infection: Rheumatoid Factor and Cryoglobulins in Different Stages of Syphilis

Auto-Immune Phenomena in Syphilitic Infection: Rheumatoid Factor and Cryoglobulins in Different Stages of Syphilis

Treatment of early syphilis with penicillin, mapharsen and bismuth (combined therapy).

IN THE PRESENT state of our knowledge the treatment of syphilis necessarily resolves itself into the treatment of the disease not as a whole but rather in its various stages. At this time the early and late stages of the disease pose different therapeutic problems. This fact becomes evident when one recalls the disparity in results now obtained by therapy in the different stages of syphilis. The problem of the successful treatment of seronegative primary syphilis can be considered solved, for no matter what schedule of intensive penicillin therapy is instituted, a satisfactory outcome can be expected, with this drug alone, in nearly 100 per cent of the patients treated. Less can be said for the treatment of the disease in the seropositive primary stage. Here the best reported results from penicillin therapy are perhaps no better than success in 92 per cent of the cases. This poorer showing is