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PubMed1980

Genetic relationship between Treponema pallidum and Treponema pertenue, two noncultivable human pathogens.

Genetic relationships among two strains of Treponema pallidum (Nichols and KKJ) and a strain of T. pertenue were determined by measuring the degree of deoxyribonucleic acid sequence homology. The results in indicated that these three virulent, noncultivable treponemes were genetically indistinguishable. Like T. pallidum (Nichols), T. pertenue (Gauthier) had no detectable deoxyribonucleic acid sequence homology with T. phagedenis (biotype Reiter), T. refringens (biotype Noguchi), or with salmon sperm.

Base Sequence↗

PubMed1988

Antigenic relatedness and N-terminal sequence homology define two classes of periplasmic flagellar proteins of Treponema pallidum subsp. pallidum and Treponema phagedenis.

The periplasmic flagella of many spirochetes contain multiple proteins. In this study, two-dimensional electrophoresis, Western blotting (immunoblotting), immunoperoxidase staining, and N-terminal amino acid sequence analysis were used to characterize the individual periplasmic flagellar proteins of Treponema pallidum subsp. pallidum (Nichols strain) and T. phagedenis Kazan 5. Purified T. pallidum periplasmic flagella contained six proteins (Mrs = 37,000, 34,500, 33,000, 30,000, 29,000, and 27,000), whereas T. phagedenis periplasmic flagella contained a major 39,000-Mr protein and a group of two major and two minor 33,000- to 34,000-Mr polypeptide species; 37,000- and 30,000-Mr proteins were also present in some T. phagedenis preparations. Immunoblotting with monospecific antisera and monoclonal antibodies and N-terminal sequence analysis indicated that the major periplasmic flagellar proteins were divided into two distinct classes, designated class A and class B. Class A proteins consisted of the 37-kilodalton (kDa) protein of T. pallidum and the 39-kDa polypeptide of T. phagedenis; class B included the T. pallidum 34.5-, 33-, and 30-kDa proteins and the four 33- and 34-kDa polypeptide species of T. phagedenis. The proteins within each class were immunologically cross-reactive and possessed similar N-terminal sequences (67 to 95% homology); no cross-reactivity or sequence homology was evident between the two classes. Anti-class A or anti-class B antibodies did not react with the 29- or 27-kDa polypeptides of T. pallidum or the 37- and 30-kDa T. phagedenis proteins, indicating that these proteins are antigenically unrelated to the class A and class B proteins. The lack of complete N-terminal sequence homology among the major periplasmic flagellar proteins of each organism indicates that they are most likely encoded by separate structural genes. Furthermore, the N-terminal sequences of T. phagedenis and T. pallidum periplasmic flagellar proteins are highly conserved, despite the genetic dissimilarity of these two species.

Amino Acid Sequence↗

PubMed1993

Polypeptides of Treponema pallidum: progress toward understanding their structural, functional, and immunologic roles. Treponema Pallidum Polypeptide Research Group.

Treponema pallidum subsp. pallidum, the spirochete that causes syphilis, is unusual in a number of respects, including its small genome size, inability to grow under standard in vitro culture conditions, microaerophilism, apparent paucity of outer membrane proteins, structurally complex periplasmic flagella, and ability to evade the host immune responses and cause disease over a period of years to decades. Many of these attributes are related ultimately to its protein content. Our knowledge of the activities, structure, and immunogenicity of its proteins has been expanded by the application of recombinant DNA, hybridoma, and structural fractionation techniques. The purpose of this monograph is to summarize and correlate this new information by using two-dimensional gel electrophoresis, monoclonal antibody reactivity, sequence data, and other properties as the bases of polypeptide identification. The protein profiles of the T. pallidum subspecies causing syphilis, yaws, and endemic syphilis are virtually indistinguishable but differ considerably from those of other treponemal species. Among the most abundant polypeptides are a group of lipoproteins of unknown function that appear to be important in the immune response during syphilitic infection. The periplasmic flagella of T. pallidum and other spirochetes are unique with regard to their protein content and ultrastructure, as well as their periplasmic location. They are composed of three core proteins (homologous to the other members of the eubacterial flagellin family) and a single, unrelated sheath protein; the functional significance of this arrangement is not understood at present. Although the bacterium contains the chaperonins GroEL and DnaK, these proteins are not under the control of the heat shock regulon as they are in most organisms. Studies of the immunogenicity of T. pallidum proteins indicate that many may be useful for immunodiagnosis and immunoprotection. Future goals in T. pallidum polypeptide research include continued elucidation of their structural locations and functional activities, identification and characterization of the low-abundance outer membrane proteins, further study of the immunoprotective and immunodiagnostic potential of T. pallidum proteins, and clarification of the roles of treponemal proteins in pathogenesis.

Amino Acid Sequence↗

PubMed2005

Treponema denticola infection is not a cause of false positive Treponema pallidum serology.

It has long been assumed that parodontal disease can be a cause of false positive results in syphilis serology, but so far there are no definitive data supporting this hypothesis. In this study we tested 250 serum samples obtained from blood donors. All of them were negative when routinely screened for antibodies against Treponema pallidum. Then, all these samples were tested by immunoenzymatic (ELISA) and Western Blot (WB) assays to investigate reactivities against T. denticola. Thirteen samples showed a strong positivity when tested by both methods. When tested by WB against T. pallidum no sample met the positivity criteria. Nevertheless, bands with molecolar weights of about 30-35 KDa (endoflagellar core antigens) were recognized. All the 13 subjects serologically T. denticola positive underwent oral clinical and radiological observation: all showed a very poor parodontal status (CPSS > 103). Eleven crevicular fluid samples out of the total of 13 patients were T. denticola positive by Real Time PCR carried out using a LightCycler system. In this study we demonstrated that the presence of T. denticola in the crevicular fluid samples obtained from patients with a severe periodontal status and/or a positive serology against T. denticola is not a cause of false positive results in syphilis serology.

Antibodies, Bacterial↗

PubMed1986

Isolation and characterization of antigens from Treponema phagedenis biotype Reiter cross-reacting with Treponema pallidum.

The specific aims of the studies reviewed here were to design a rational purification strategy for Reiter treponeme antigens using combinations of different chromatographic principles, to isolate and characterize the Reiter treponemal antigens, especially the antigens labelled TR-b, TR-c, TR-dl, TR-d2, and TR-e cross-reacting with antigens in Treponema pallidum, to develop and evaluate the use of these purified antigens in syphilis diagnostic enzyme-linked immunosorbent assays (ELISA), to investigate whether Reiter treponeme antigens can induce protective immunity against experimental syphilis in rabbits. By combining anion exchange chromatography, gel filtration, agarose gel electrophoresis, hydrophobic interaction chromatography, and affinity chromatography on Iminodiacetic acid-Sepharose CL 4B and Lysine Sepharose 4B we were able to isolate seven different water soluble Reiter antigens from one single Reiter sonicate supernatant (I, II, III, IV, V, VI). The applied chromatographic matrices were selected due to their efficiency for separation of the Reiter antigens in pilot experiments. In addition to the above mentioned Reiter antigens additional antigens labelled TR-o (V) and LPS (VI) were isolated. TR-b, TR-c, and TR-o were shown to be protein antigens (III, IV, V). The TR-c antigen of the Reiter treponeme cross-reacted not only with an antigen in T. pallidum but also with an antigen common to a wide range of bacteria (IX). The TR-d antigen composed of a ribonucleic acid component (TR-dl) (II) and a protein component (TR-d2). The TR-e antigen represented the flagellum of the bacterium (I), and the LPS antigen was a pure lipopolysaccharide antigen (VI).(ABSTRACT TRUNCATED AT 250 WORDS)

Antigens, Bacterial↗

PubMed2004

Evaluation of a Treponema pallidum-specific IgM enzyme immunoassay and Treponema pallidum western blot antibody detection in the diagnosis of maternal and congenital syphilis.

BACKGROUND: Congenital syphilis (CS) is a result of untreated or inadequately treated maternal syphilis. CS is more likely with early stages of maternal syphilis, but most mothers lack signs or symptoms and the risk of CS is unclear. GOAL: The goal of this study was to evaluate Treponema pallidum IgM Western blot (TP IgM WB) and a T. pallidum IgM enzyme immunoassay (TP IgM ELISA) in mothers with syphilis to determine if positive tests better indicate a risk of CS than a rapid plasma reagin titer >/=1:16. STUDY DESIGN: Ninety-seven mother-baby pairs with reactive syphilis serology were evaluated. RESULTS: TP IgM WB tests were positive in 18 pregnancies (7 of 18 babies had CS) and negative in 79 pregnancies (7 of 82 babies had CS). Thirty-two mothers had titers >/=1:16 (6 babies with CS) and 65 mothers had titers </=1:8 (8 babies with CS). CONCLUSION: TP IgM tests better identify mothers at risk of delivering babies with CS than maternal titer >/=1:16.

Adult↗

PubMed2001

Proteolytic activity among various oral Treponema species and cloning of a prtP-like gene of Treponema socranskii subsp. socranskii.

The proteolytic activity of 11 treponemal strains representing different phylogenetic groups was investigated by SDS-polyacrylamide gel electrophoresis with copolymerised casein, gelatin and fibrinogen as substrates. The activity was specified to be trypsin- and chymotrypsin-like by the cleavage of synthetic substrates BAPNA and SAAPFNA, respectively. Nine strains degrade casein and the synthetic substrate BAPNA. Chymotrypsin-like activity specifically inhibited by phenylmethylsulfonyl fluoride was found in four treponemes. Southern blot analysis using a Treponema socranskii subsp. socranskii-specific prtP probe confirmed the presence of prtP homologous genes in these four strains. The internal fragments of the chymotrypsin-like protease genes were cloned and sequenced after PCR amplification. Here we report the cloning of the complete prtP-like gene of T. socranskii subsp. socranskii, an organism shown to possess epidemiologic relevance in periodontitis.

Blotting, Southern↗

PubMed1988

Lipooligosaccharides from Treponema hyodysenteriae and Treponema innocens.

Lipooligosaccharides from Treponema hyodysenteriae serotypes 1 through 7, attenuated T. hyodysenteriae serotypes 1 and 2, and five strains of T. innocens were extracted with hot phenol water. The extracts were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis separation and analyzed by lipopolysaccharide selective silver staining and Western blot (immunoblot) immunodetection. Silver staining revealed the presence of two bands that ranged between 18,000 and 24,000 daltons and that were serotype specific for T. hyodysenteriae. Attenuation of pathogenic strains resulted in the loss of the higher-molecular-weight band. Four of five T. innocens strains also lacked this particular band. T. innocens 421 had six bands between 17,000 and 26,900 daltons. Western blots with hyperimmune rabbit sera and convalescent-phase swine sera revealed antigenic variation among serotypes of T. hyodysenteriae and attenuated serotypes of T. hyodysenteriae. Convalescent-phase swine sera failed to recognize lipopolysaccharides from T. innocens. Differences in results obtained by lipopolysaccharide selective silver staining versus immunoblotting of the lipopolysaccharide preparations probably indicate that these two methods identify separate characteristics of the same molecule.

Animals↗

PubMed2003

Antibody responses elicited against the Treponema pallidum repeat proteins differ during infection with different isolates of Treponema pallidum subsp. pallidum.

Variation in the expression of the different Tpr proteins in the syphilis spirochete, Treponema pallidum subsp. pallidum, may have important implications in its ability to evade host immune detection and cause persistent infection. In the present study we examined the pattern of antibody responsiveness to different Tpr members during infection with three isolates of T. pallidum. There was variability in the specificities and temporal patterns of reactivity of the antibodies elicited against the individual Tpr proteins, suggesting that isolates may express different repertoires of Tpr proteins during infection.

Amino Acid Sequence↗

PubMed1980

Treponema innocens lipids and further description of an unusual galactolipid of Treponema hyodysenteriae.

The lipids of Treponema innocens, type strain B256, formerly considered a nonpathogenic isolate of T. hyodysenteriae, have been analyzed and compared with the lipids of T. hyodysenteriae. The lipids of T. innocens comprised 16% of the cell dry weight. Polar lipids amounted to about two-thirds of the total lipids and consisted of 61.9% phospholipids and 38.1% glycolipid. Neutral lipids consisted mainly of sterols. The phospholipids were principally phosphatidylglycerol, phosphatidylcholine, and cardiolipin. Minor amounts of lysophosphatidylcholine, sphingomoyelin, and a relatively nonpolar, unidentified phospholipid were present. The latter lipid has not been detected in T. hyodysenteriae. The glycolipid fraction of T. innocens contained a single component, monoglucosyldiglyceride, in contrast to the occurrence in T. hyodysenteriae of two components: monogallactosyldiglyceride and a less-polar glycolipid tentatively identified as acylmonogalactosyldiglyceride (the additional acyl moieties being 86.6% acetyl, 11.6% propionyl, and 1.6% n-butyryl groups). Alk-1-enyl ether analogs comprised 24.6% of the total phospholipids and glycolipid of T. innocens, or about one-third of the amount in T. hyodysenteriae. The acyl and alk-1-enyl moieties of T. innocens consisted of greater than or equal to 92% of 14:0, iso-15:0, and 16:0 chains. In contrast to T. hyodysenteriae, anteiso-15:0 moieties were not detected, and a reversed distribution of 14:0 and iso-15:0 alk-1-enyl moieties occurred in the two species.

Galactolipids↗

PubMed2000

Multiple alleles of Treponema pallidum repeat gene D in Treponema pallidum isolates.

Two new tprD alleles have been identified in Treponema pallidum: tprD2 is found in 7 of 12 T. pallidum subsp. pallidum isolates and 7 of 8 non-pallidum isolates, and tprD3 is found in one T. pallidum subsp. pertenue isolate. Antibodies against TprD2 are found in persons with syphilis, demonstrating that tprD2 is expressed during infection.

Alleles↗

PubMed1996

Detection of pathogen-related oral spirochetes, Treponema denticola, and Treponema socranskii in dental plaque from dogs.

Spirochetes have been observed in dental plaque from dogs, but specific spirochetes have not been identified. In particular, it is not known whether treponemes associated with periodontal diseases in humans also occur in dogs, and whether, like in humans, detection of specific treponemes correlates with periodontal status of dogs. Forty-two dogs were grouped according to the worst periodontal condition in the mouth, as determined by overt signs of inflammation and pocket probing depths. A representative specimen of dental plaque was obtained by pooling subgingival plaque collected from three uniform reference sites, irrespective of periodontal status at selected sites. The presence of pathogen-related oral spirochetes. Treponema denticola, and T. socranskii was determined using specific monoclonal antibodies in an immunocytochemical microscopic assay. All three treponemes were detected in all groups, but a significantly greater proportion of dogs with pocket probing depths > or = 5 mm had detectable treponemes, compared to dogs that were in periodontal health.

Animals↗

PubMedDate not supplied