Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “TREPONEMA”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 991 records · Page 55Linked to original sources

Optimum concentration of dissolved oxygen for the survival of virulent Treponema pallidum under conditions of low oxidation-reduction potential.

A maintenance medium with a low oxidation-reduction (redox) potential, when gently bubbled with 5% oxygen in nitrogen or with air for various periods of time, gave a range of dissolved oxygen concentrations between 1.6 and 5.8 micrograms/l. Virulent Treponema pallidum (Nichols strain) inoculated into these media were assayed 24 and 48 hours later for motility and virulence and were compared with samples taken at zero time. Virulent T. pallidum survived best in the presence of 2.4 micrograms/l dissolved oxygen over a 48-hour period, which corresponded to a gaseous mixture of 3% oxygen in nitrogen. Higher concentrations of oxygen did not give significantly different results from anaerobic conditions over this period. Thus, until it can be grown in vitro, T. pallidum would appear to be a microaerophilic bacterium.

Anaerobiosis↗

Limited protection of rabbits against infection with Treponema pallidum by immune rabbit sera.

After intradermal infection of rabbits with 3 x 10(6) Treponema pallidum (Melbourne 1 strain) samples of serum were taken at one, two, three, four, and six months after infection. Normal rabbits were passively immunised with these sera, challenged with intradermal doses (10(4), 10(3), 10(2), 10) of T. pallidum, and the latent periods of infection, lesion diameters, and the number of inoculation sites developing into lesions were observed. The sera taken at three, four, and six months reduced the number of intradermal inoculation sites that developed into syphilitic lesions after challenge with 10 T. pallidum. These same three sera also increased the latent period of infection after challenge with 10(4) T. pallidum. The transfer of 50 ml of immune serum per rabbit over a nine-day period before challenge had very little effect on the course of the challenge infection. Only a low level of immunity in rabbits to this strain of T. pallidum appears to be mediated by immune serum but this small degree of protection did increase with time after infection. Enhanced growth of T. pallidum in the serum-recipient rabbits did not occur, thus suggesting that none of the sera was immunosuppressive.

Animals↗

Sulfhydryl oxidation using procedures and experimental conditions commonly used for Treponema pallidum.

Certain reducing agents containing sulfhydryl groups are important to the in-vitro survival of Treponema pallidum. Discrepancies occur, however, concerning the agents and the concentrations that are optimal. To clarify some of this confusion, sulfhydryl oxidation was determined using procedures and experimental conditions commonly used for T pallidum. Sulfhydryl oxidation varied according to the type of culture medium, the size of the culture vessels, the volume of the culture medium, and the gaseous environment within the culture vessels, as well as the method of extracting treponmes from infected testicular tissue. Dithiothreotol maintained highly reduced conditions by reducing disulfide groups to sulfhydryl groups. Lastly, the organisms influenced the sulfhydryl concentration by either direct oxidation or specific uptake. The sulfhydryl content was sharply decreased in the presence of viable preparations of T pallidum compared with heated preparations or membrane filtrates of viable preparations.

Animals↗

Pleural effusion disease agent as passenger of Treponema pallidum suspensions from rabbits. Survey of laboratories.

Material from rabbits used for the propagation of Treponema pallidum in 12 selected laboratories was examined for a viral passenger agent of the treponemal suspensions. An agent which causes clinical or subclinical pleural effusion disease (PED) in rabbits and which is serologically identical or related to the PED agent isolated in Copenhagen was found as a contaminant in treponemal suspensions in laboratories in Europe, the USA, and Japan. The experience gained in the Scandinavian treponematoses laboratories suggests that strains of T pallidum contaminated with the PED agent can be purified by passage through hamsters.

Animals↗

Quantitative microhaemagglutination assay for Treponema pallidum antibodies in experimental syphilis.

The quantitative microhaemagglutination assay for Treponema pallidum antibodies (MHA-TP) was studies in 52 untreated and treated rabbits with experimental syphilis. Rabbits with incubating experimental syphilis were cured or inadequately treated with penicillin G and some cured rabbits were later reinfected. MHA-TP conversion occurred within 45 days in untreated rabbits. Titres reached peak levels about four months after inoculation and remained relatively high for up to two years. The quantitative MHA-TP test differentiated between rabbits cured of experimental incubating syphilis and those untreated and inadequately treated. MHA-TP titres decreased after treatment given six or 12 months after inoculation but reversion did not occur. MHA-TP conversion or significant increases in titre occurred as soon as seven days after reinfection and preceded corresponding changes in a quantitative non-treponemal test. The MHA-TP is useful as a screening test for treponemal antibodies in rabbits. The quantitative MHA-TP in humans after treatment for syphilis and reinfection deserves further study.

Animals↗

Experimental infection of man with rabbit-virulent Treponema paraluis-cuniculi.

Virulent Treponema paraluis-cuniculi was inoculated intradermally into the arm of a human volunteer and into the shaved backs of 10 rabbits. An identical, but heat-killed, preparation was inoculated into the opposite arm of the volunteer as control. A superficial and transient infection developed in the volunteer, shown by a small zone of erythema that persisted for 24 days. The control preparation caused a smaller zone of erythema that disappeared after five days. A very poor immune response was detected by standard serological tests for syphilis. The inoculated rabbits developed lesions about six days after infection and seroconverted by 84 days. The poor antitreponemal antibody response to T paraluis-cuniculi infection in the volunteer suggests that this naturally attenuated treponeme may not be suitable as a vaccine against infection with t pallidum in humans.

Animals↗

Specificity of the Treponema pallidum haemagglutination test. Analysis of results.

The automated haemagglutination assay using Treponema pallidum antigen (AMHA-TP) and the Venereal Disease Research Laboratory (VDRL) test were used to examine 330 163 sera. Reactive results were checked by the fluorescent treponemal antibody-absorption (FTA-ABS) test. When isolated reactivity or non-reactivity in the AMHA-TP test was investigated an estimated margin of error of 0.7% probably wrongly non-reactive and 0.008% presumably false non-reactive results were found. These figures were confirmed by randomised FTA-ABS tests on 504 sera with repeat AMHA-TP tests. The latter is therefore still the most reliable and practicable method for mass screening for syphilis.

Antibodies, Bacterial↗

Redox potential and survival of virulent Treponema pallidum under microaerophilic conditions.

A strongly reduced culture medium, capable of maintaining the virulence of Treponema pallidum (Nichols) for several days, was exposed to an atmosphere of 3% oxygen in nitrogen for 2-3 days before inoculation with T pallidum. By using various volumes of medium in uniform tubes a range of redox potentials (Ecal) from -94 mV to -325 mV was produced depending on the surface area-to-volume ratios of the medium. The anaerobic medium had an Ecal value of -387 mV. The medium was inoculated with T pallidum and incubated in an atmosphere of 3% oxygen. The survival of treponemes at different redox potentials was monitored by observing the retention of motility and by measuring the latent period of infection after inoculation of the cultures into the shaved backs of rabbits. Under these conditions T pallidum survived longest at low (electronegative) redox potential. An inverse linear relationship was observed between the redox potential of the culture medium and the survival of T pallidum, as measured by the time required for a 90% reduction of virulent organisms. No optimum redox potential was detected, the most electronegative medium (-325 mV, Ecal) giving the best survival.

Bacteriological Techniques↗

Analysis of serum IgG against Treponema pallidum protein antigens in experimentally infected rabbits.

Sensitive and highly specific radioimmunoprecipitation and gel electrophoreses-fluorography were used to monitor the serum IgG response of experimentally infected rabbits against key protein antigens of Treponema pallidum. The sera from six rabbits challenged intradermally (ID) and six intratesticularly (IT) with T pallidum were examined over a period of four months. A specific antibody response to most treponemal surface proteins (including those ligands implicated as mediating host-cell-surface adherence) developed by about day 20 after inoculation. Maximum antibody concentrations against most of these immunogens were detected by day 50 in all animals irrespective of the route of inoculation. Differences as well as similarities in the IgG response to infection were observed among selected rabbits. Furthermore, the antibody reactivity of each animal was comparable to that of human sera from patients with secondary syphilis. Antibody directed against treponemal outer envelope proteins was detected by preferential agglutination of washed radiolabelled organisms with protein A-bearing Staphylococcus aureus in the presence of syphilitic rabbit sera.

Animals↗

Morphological destruction of cultured cells by the attachment of Treponema pallidum.

The incubation of Treponema pallidum with rabbit testicular cells, HEP-2 cells, human foreskin cells, rat cardiac cells, and rat skeletal muscle cells caused morphological disruption of these cultured cells. Control preparations of heat-inactivated treponemes, a high-speed supernatant in which treponemes had been pelleted, and culture medium failed to damage the tissue cells, as did viable treponemes when the cells were incubated in inverted Sykes-Moore chambers. Thus, cellular disruption is not associated with soluble treponemal, soluble inflammatory, or soluble testicular constituents but is mediated by the specific attachment of T pallidum. This organism apparently elaborates some type of toxic activity that lyses membranes: this may explain some of the histopathology of syphilitic disease.

Animals↗

Effects of anaerobic and microaerophilic conditions of extraction and incubation on the survival of Treponema pallidum in vitro.

Treponema pallidum extracted from infected rabbit testes under anaerobic conditions survived longer in vitro than those extracted under aerobic conditions. Anaerobically extracted treponemes were incubated anaerobically for 0, 12, 24, 36, or 48 hours and then exposed to microaerophilic conditions (3% oxygen) for further incubation. Treponemes transferred to microaerophilic conditions after 36 or 48 hours' anaerobic incubation maintained significantly greater viability compared with those kept under constant microaerophilic conditions, although there was no difference after 12 or 24 hours. T pallidum incubated under constant anaerobic conditions, however, usually maintained greater viability than those kept under constant microaerophilic conditions. These results suggest that T pallidum is sensitive to oxygen toxicity both during initial extraction from orchitic rabbit testes and subsequent incubation in vitro. In the latter case, it can be partially protected by a period of anaerobic incubation in vitro, before exposure to microaerophilic conditions.

Anaerobiosis↗

Interferon in rabbit sera after inoculation with Treponema pallidum suspensions contaminated with PED virus.

Paired rabbits sera were examined for the presence of interferon and pathogenicity for rabbits. The sera were obtained before and 48 hours after inoculation with Treponema pallidum suspensions of rabbit origin in 12 selected laboratories. Classical interferon, detectable in dilutions from 1/9 to 1/81, were found in 27 out of 39 postinoculation sera from which the pleural effusion disease (PED) virus was isolated. Serum interferon was not detectable in dilutions greater than or equal to 1/9 in 16 virus-negative postinoculation sera or in any of the 55 preinoculation sera. Interferon was found more often in sera from which highly virulent strains of PED virus were isolated than in sera from which strains of low virulence were isolated. The serum interferon assay provides useful presumptive evidence of contamination of rabbit-passaged treponemes with PED virus, but the assay is least useful when PED virus is present subclinically.

Animals↗

Evaluation of the role of the Treponema pallidum immobilisation test in Britain.

After specimens of sera from 100000 patients had been screened by a reagin and Treponema pallidum haemagglutination assay (TPHA) 2.7% required further examination. The fluorescent treponemal antibody-absorption (FTA-ABS) test confirmed either a negative or positive TPHA result in 95% of the samples, so that no further investigations were required. This accounted for 99% of the specimens submitted to the laboratory. Repeat tests on a further sample resolved many of the remaining discrepancies. Only 0.05% of patients, in whom repeat tests confirmed a positive TPHA but a negative FTA-ABS result, benefited from a TPI test.

Fluorescent Antibody Technique↗

Scanning electron microscopy of the attachment of Treponema pallidum to nerve cells in vitro.

Treponema pallidum (Nichols strain) was incubated with cultured nerve cells derived from rat embryos. Primary cultures were established from dorsal root ganglia, superior cervical ganglia, and spinal cord. Using phase contrast microscopy treponemes were seen to interact with the nerve cells in a similar manner to other cultured mammalian cells. Organisms began to attach within minutes after inoculation, actively motile organisms attached at the tip of one end, higher numbers of organisms attached with continued incubation, and attached organisms survived longer than unattached organisms. T pallidum attached both to nerve cell bodies and to neuronal processes of each of the three nerve cell cultures. As shown by scanning electron microscopy the mechanism of attachment was identical to that of cultured cells derived from rabbits testis, rat skeletal muscle, and human cervical carcinoma. There was no indentation or swelling of the cultured cell surface at the point of attachment, just a close physical proximity of organisms and cells. These techniques provide a biological means of studying the in-vitro detrimental influences of micro-organisms on nerve tissue.

Animals↗

Electrophysiological dysfunction and cellular disruption of sensory neurones during incubation with Treponema pallidum.

Treponema pallidum (Nichols strain) was incubated with cultured nerve cells derived from dorsal root ganglia of rat embryos. The electrophysiological response of these neuronal cells was then investigated. Cells exposed to 2 X 10(8) treponemes/ml responded abnormally after 13 hours and failed to respond after 18 hours. In contrast, control preparations exposed to heat-inactivated treponemes or to culture medium responded normally after 72 hours. Extended incubation with viable treponemes resulted in various degrees of nerve cell disruption as shown by scanning electron microscopy. With some cells holes in the cytoplasmic membrane were detected; with others a coagulated matrix of apparent nuclear material and remnants of cytoskeletal elements indicated more severe destruction. These findings may explain the painless nature of many of the clinical manifestations of syphilis as well as the severe damage to central nervous system tissue in tertiary and congenital syphilis.

Action Potentials↗

Comparison of the morphology, antigenicity, and pathology of Nichols and Birmingham isolates of Treponema pallidum.

A strain of Treponema pallidum isolated from a homosexually acquired penile chancre in Birmingham in 1979 was serially passaged in rabbits and compared with the Nichols strain. The two strains appeared identical morphologically, and no gross differences were detected on antigenic analysis by two-dimensional immunoelectrophoresis. The Birmingham strain was, however, considerably less virulent for rabbits; the number of organisms required for equivalent time of development and severity of skin lesions was 10-fold to 100-fold greater for the Birmingham than for the Nichols strain. The pathology of testicular infection was broadly similar for both strains.

Animals↗

Factors affecting the attachment of Treponema pallidum to mammalian cells in vitro.

Attachment of Treponema pallidum (Nichols) to mammalian cells is probably the first step in the pathogenesis of syphilis. It may also be important for the multiplication of T pallidum in vitro. When factors affecting the attachment of T pallidum to mammalian cells in vitro were studied significantly greater numbers of treponemes were found to attach to baby rabbit genital organ (BRGO) cells than to five other mammalian cell lines. When attached to BRGO cells T pallidum survived longer in vitro than unattached treponemes. Eagle's minimal essential medium was superior to three other culture media in increasing attachment and maintaining the survival of treponemes. Dithiothreitol (0.25-1.0 mmol/l) had no effect on the attachment of T pallidum to BRGO cells. Anaerobic conditions were superior to microaerophilic conditions, and the latter were superior to aerobic conditions for the attachment and survival of T pallidum to BRGO cells. Within the range of concentrations tested the number of treponemes attached to the BRGO cells was directly dependent on the concentrations of viable treponemes in the inoculum. Greater numbers of treponemes attached to actively metabolising BRGO cells than to quiescent or slowly growing cells.

Animals↗

Enzyme-linked immunosorbent assay for the detection of serum antibody to outer membrane proteins of Treponema pallidum.

A highly sensitive enzyme-linked immunosorbent assay was used for the analysis of serum IgG reactivity against specific immunogenic Treponema pallidum proteins. Outer membrane treponemal proteins purified by preparative SDS-polyacrylamide gel electrophoresis were used as antigenic probes at concentrations as low as 100 ng per ml (5 ng per well in microtitre plates). Detection of anti-treponemal antibody was possible using rabbit syphilitic sera diluted to 1/10 000. The sensitivity of the assay was equal to or greater than that detected by radioimmuno-precipitation combined with gel electrophoresis and fluorography techniques and was capable of monitoring host IgG responses throughout the progress of the disease.

Animals↗