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1.  Inhibitory effect of syphilitic rabbit serum on DNA synthesis in rabbit cells in vitro. 
A previously described toxic factor associated with Treponema pallidum (Nichols) and found in extracts of syphilitic rabbit testes has now also been detected in syphilitic rabbit serum. The toxic factor, which inhibits DNA synthesis in baby rabbit genital organ (BRGO) cells in vitro, is present in rabbit serum up to 30 days after infection with T pallidum.
PMCID: PMC1046339  PMID: 6386096
2.  Susceptibility of Treponema pallidum to the toxic products of oxygen reduction and the non-treponemal nature of its catalase. 
We examined the sensitivity of Treponema pallidum (Nichols strain) to toxic products of oxygen reduction. T pallidum was sensitive to hydrogen peroxide at concentrations similar to those to which obligate anaerobes are sensitive. Accelerated death of T pallidum occurred at hydrogen peroxide concentrations below 50 mumol/l. Agents protective against hydrogen peroxide and the hydroxyl free radical (catalase, peroxidase, and mannitol) significantly enhanced treponemal survival in vitro under all three conditions of aerobiosis tested--that is, air, 3% oxygen, and 3% oxygen in conjunction with a reduced medium. Superoxide dismutase (which provides protection against superoxide radicals) did not enhance treponemal survival in normal media. When superoxide radicals were generated in the medium by means of a xanthine and xanthine oxidase system, however, the enzyme did protect T pallidum. A possible toxic involvement of singlet oxygen was also indicated by enhanced treponemal survival in air in the presence of histidine. Extracts of T pallidum from infected rabbit testes showed catalase activity which, on polyacrylamide gel electrophoresis, had the same relative mobility as purified rabbit catalase. The treponemal catalase activity was neutralised by anti rabbit catalase antiserum (raised in guinea pigs). This confirmed that the catalase was of rabbit origin and not an endogenous enzyme of T pallidum. We discuss the relation of these results to the obligate parasitism of T pallidum.
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PMCID: PMC1046263  PMID: 6421449
3.  Effect of syphilitic rabbit sera taken at different periods after infection on treponemal motility, treponemal attachment to mammalian cells in vitro, and treponemal infection in rabbits. 
The time course of antibody synthesis during syphilis was studied in experimentally infected rabbits. A rapid antibody response was seen; the rabbits became positive in both the rapid plasma reagin (RPR) test and Treponema pallidum haemagglutination assay (TPHA) by nine days after infection. Treponemal immobilising antibodies were also seen as early as nine days after infection. Antibody inhibition of treponemal attachment to baby rabbit genital organ (BRGO) cells in culture occurred with immune sera taken 30 days after infection but not earlier. When T pallidum was mixed with immune syphilitic rabbit sera taken at different stages of the infection and used to infect normal rabbits the rabbits became partially resistant to T pallidum only when the treponemes were mixed with sera taken at least 30 days after syphilitic infection. This appearance correlated well with the development of antibodies which blocked attachment of T pallidum to host cells. These antibodies may be involved in the resistance to reinfection which develops in syphilis as the disease progresses.
PMCID: PMC1046187  PMID: 6347332
4.  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.
PMCID: PMC1046124  PMID: 6337680
5.  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.
PMCID: PMC1045951  PMID: 7028206
6.  Sequential changes in susceptibility to Treponema pallidum of rabbits previously infected with Treponema paraluis-cuniculi 
Rabbits immunised with virulent Treponema paraluis-cuniculi were challenged intradermally with graded doses of Treponema pallidum at three, five, seven, 12, and 30 months to ascertain the level of protection to T pallidum at various intervals after immunisation.
Rabbits challenged at three months after immunisation showed no protection against T pallidum and developed syphilitic lesions significantly faster than the control rabbits, which suggests that the former rabbits were immunosuppressed. Some protection was evident at five and seven months after immunisation, as fewer inoculation sites developed syphilitic lesions with challenges of 103, 102, and 10 T pallidum and lesions developed significantly slower with 106 challenge. Two rabbits showed significant protection at 12 months after immunisation but a third, presumably still immunosuppressed, developed lesions significantly faster than the control rabbits after challenge. At 30 months after immunisation one rabbit was completely protected and developed no lesions after challenge; the other rabbit showed only partial protection against challenge with 104, 103, and 102 but complete protection against challenge with 10 T pallidum.
T paraluis-cuniculi appeared to induce a state of immunosuppression by three months after infection; in one rabbit this may have been 12 months. In most immunised rabbits, however, limited cross-protection to low challenge doses of T pallidum developed by five months and was also detectable at seven and 12 months. Only one rabbit was completely resistant to challenge with 104T pallidum after 30 months and another was only partly immune. Thus, T paraluis-cuniculi infection does not produce a rapid pronounced cross-immunity to T pallidum in rabbits, which may thus limit its usefulness as a vaccine against syphilis.
PMCID: PMC1045858  PMID: 7470830
7.  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.
PMCID: PMC1045857  PMID: 7470837
8.  Susceptibility of rabbits venereally infected with Treponema paraluis-cuniculi to superinfections with Treponema pallidum. 
Three female rabbits, venereally infected with Treponema paraluis-cuniculi between five and eight months previously, had treponeme-containing genital lesions and positive results to serological tests for syphilis. These rabbits and four normal female rabbits were challenged with Treponema pallidum intradermally on the shaved back with triplicate doses of 10(4), 10(3), 10(2), and 10 treponemes (all at different sites). Significantly fewer syphilitic lesions developed in the rabbits previously infected with T paraluis-cuniculi (4/36) compared with the control rabbits (37/48), and the mean size of the lesions (diameter of induration) was significantly less (4 mm compared with 10 mm). It appears that previous venereal infection of female rabbits with T paraluis-cuniculi induced a level of cross-immunity against infection with T pallidum. Protection was however not complete.
PMCID: PMC1045837  PMID: 7004567
10.  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.
PMCID: PMC1045696  PMID: 393362
11.  Susceptibility of rabbits to Treponema pallidum after infection with Mycobacterium bovis. 
Rabbits stimulated with Mycobacterium bovis (strain BCG) one month before challenge with Treponema pallidum (Nichols) did not show any modification in their development of syphilitic lesions. A second infection with BCG given at the same time and at the same intradermal site as the T. pallidum challenge also failed to protect the rabbits against syphilis. Thus the non-specific activation of cell-mediated immunity by BCG does not appear to protect rabbits against T. pallidum infection even when both activation and challenge take place in the dermis. The role of the macrophage in syphilis remains obscure.
PMCID: PMC1045695  PMID: 393361
12.  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.
PMCID: PMC1045694  PMID: 393360

Results 1-13 (13)