Reiter Protein Research Articles

Autolymphocytotoxins in syphilis.

Nineteen of 23 serum samples from patients with secondary syphilis, six of 24 from patients with primary syphilis, and four of seven from patients with early latent syphilis had complement-dependent autolymphocytotoxic reactivity at 15 degrees C. After treatment, the reactivity was decreased. It was not directed only against autologous lymphocytes but also against allogeneic lymphocytes. Sephadex G-200 gel filtration showed that the lymphocytotoxins were present in the IgM-containing exclusion peak. Furthermore, serum samples that had high antibody titres in the Reiter protein complement-fixation test or Wassermann-Kolmer reaction also had a high frequency of cold-reacting autolymphocytotoxins. This indicates that the autolymphocytotoxic reactivity is related to the humoral response in syphilis.

Sexually transmitted diseases in clinic patients in Lagos.

In a study of patients attending an STD referral clinic in Lagos, Nigeria, the age distribution was similar to that seen on other parts of the world and the male-to-female ratio was 1.3:1. Non-specific genital infection was diagnosed in about 59% of patients and gonorrhoea in 19%. Most strains of Neisseria gonorrhoeae were sensitive to penicillin (1 unit) and the six relatively penicillin-resistant strains did not produce penicillinase. Candidosis and trichomoniasis were each diagnosed in about 10% of patients and latent syphilis in 1.4% (four cases). Genital warts were also uncommon; venereophobia and other conditions accounted for 1% and 2.2% of cases respectively.

Development of of macrophage migration inhibition in rabbits infected with virulent Treponema pallidum

Peritoneal exudate cells from rabbits infected with Treponema pallidum Nichols were used as indicators of macrophage migration inhibitory factor activity. Between 3 and 15 weeks after infection, the migration of peritoneal exudate cells was inhibited in the presence of 3 to 25 microgram of T. phagedenis biovar Reiter protein per ml. Before this period, the migration patterns of peritoneal exudate cells from infected animals were uninhibited and similar to those from noninfected control rabbits. These observations were correlated with the development of active cell-mediated immunity during experimental T. pallidum infection.

Secondary Syphilis and Sarcoidosis

To the Editor.— A case of secondary syphilis and sarcoidosis, which was diagnosed radiologically, was reported recently by McMillan and Burns ( Arch Dermatol 111:264, 1975). We would like to report a similar case. The patient, born in 1932, was seen in the clinic of the Department of Ophthalmology, University of Geneva, in February 1971 for uveitis. One year before coming to the clinic, he had had a chancre of the foreskin and urethritis; he was unable to specify the treatment given at that time. Approximately six months later, spreading, erythematous lesions appeared on the torso. When he came to the clinic of the Department of Dermatology in July 1971, acuminate, papular lesions, scattered all over the torso, and epitrochlear nodes had been developing for three months. The following laboratory tests for syphilis were reactive: Bordet-Wassermann, + + +; Reiter protein complement fixation, + + +; VDRL test for syphilis (initially with a titer of 1/8; later,

Comparison of the counter-immunoelectrophoresis technique with the Reiter protein and three other serological tests as a first line test for syphilis

The counter-immunoelectrophoresis technique with the Reiter protein (RP-CIE) was compared with two complement fixation tests (Kolmer and RPCF) and a flocculation test (VDRL) in sensitivity and specificity. Of the 1,927 consecutive attendants of a venereal disease clinic whose serum samples were used, 250 were considered to be syphilitic. The number of true-positive and false-positive reactions were: 121 and 4 (VDRL), 124 and 2 (Kolmer), 179 and 41 (RPCF), 166 and 16 (RP-CIE). The VDRL and the RPCF combined were more sensitive and less specific than the VDRL combined with the Kolmer. If the RPCF was replaced by the RP-CIE the sensitivity remained the same but the specificity was higher. The RP-CIE scored more positives than the RPCF in untreated cases of primary syphilis. The results argue for substitution of the RP-CIE for the Kolmer in the combination with the VDRL in the serodiagnosis for syphilis. Moreover, the RP-CIE presents the technical advantages of simplicity, speed of performance, and of not being hampered by the anticomplementary nature of the serum sample.

Isolation and antigenic characteristics of axial filaments from the Reiter Treponeme.

Axial filaments were isolated and purified from Reiter treponemes after detergent solubilization of the cells" outer envelope. The axial filaments were separated from the spirochetal cells by shearing, purified by density gradient centrifugation, and fragmented by ultrasonication. Acrylamide gel electrophoresis of dissociated filaments revealed two major protein bands. Gel diffusion precipitin tests and immunoelectrophoresis between a purified axial filament suspension and anti-Reiter treponeme serum gave a single precipitin line. Checkerboard complement fixation tests also gave results consistent with a single antigen-antibody system. Tests with immune sera to other cultivable spirochetes were positive with some and negative with others. In addition, strongly positive reactions were obtained in complement fixation and precipitin tests with sera from rabbits and humans with syphilis and other treponematoses. However, both serological tests gave reactions of partial identity between the antigen(s) of Reiter treponeme axial filaments and those of the pathogenic treponemes. It was concluded from these studies that the axial filaments were probably the cellular locus of the so-called "Reiter protein" antigen of syphilis serology.

Spirochetes in Late Seronegative Syphilis, Penicillin Notwithstanding.

Before the advent of serological tests for syphilis, the disease was difficult to diagnose. When Wasserman, Hinton, and others developed their serological tests, the diagnostic problem was simplified. With the development of Reiter protein complement fixation (RPCF), fluorescent treponemal antibody (FTA), Treponema pallidum immobilization (TPI), and fluorescent treponemal antibodyabsorption (FTA-ABS) tests, both the specificity and sensitivity have been greatly improved. What is most conclusive, however, is the demonstration of Treponema pallidum , not only during the early stages but also at the late stage of the disease. With the advent of penicillin, treatment of syphilis has become a simple matter. Nowadays, syphilis is considered as just another infectious disease, no longer taught separately as syphilology. Nonetheless, in this book, J. L. Smith takes the opposite view: syphilis is now both hard to diagnose and hard to treat. Section 1 of the book reviews the problem of late seronegative (by reaginic tests)

La réaction d’hémagglutination passive pour la syphilis (antigène Reiter)

Formaline-alcohol fixed sheep erythrocytes are sensitized with sonified Reiter protein antigen under different conditions. It is found that after 3 months storage a factor appears in the antigen, which is probably identical with the factor responsible for the precipitation of the serum proteins in abandoned cultures and which we therefore call ‘seroprecipitating factor’. Due to this factor, erythrocytes sensitized with old batches of antigen agglutinate in normal serum. As this factor has a much higher affinity for tannic acid than for bis-diazobenzidine, tannic acid is less suitable as a coupling agent than bis-diazobenzidine. When the erythrocytes are sensitized with freshly prepared antigen by means of bis-diazobenzidine, they are not agglutinated by normal serum nor by ‘biologic false positive’ sera. With these erythrocytes the syphilitic sera used for this study, showed agglutinating titers ranging from 1 : 20 to 1 : 320. When the cells are treated separately with bis-diazobenzidine first and antigen later, they are also suitable for a rapid slide hemagglutination test. The sensitized erythrocytes can be lyophilized. As lyophilization provokes a loss of antigenicity, the sensitization has then to be done with twice the normal concentration of antigen. The best results are obtained when all the constituents of the sensitized cell suspension are ten times concentrated.

La réaction d’hémagglutination passive pour la syphilis (antigène Reiter)

The indirect hemagglutination reaction (IHR) is compared with the classical tests and with the immunofluorescence test (FTA 200) by means of 50 syphilitic sera and 7 ‘biologic false positive’ sera (BFP). The results obtained suggest that (a) the IHR may become positive earlier than the Reiter protein complement fixation test (RPCF) and the FTA 200, (b) the IHR negativates more often after adequate treatment then the RPCF and the FTA 200, (c) the IHR is mainly positive during the evolutive phases of the disease, (d) the titers obtained in recent primary disease and treated secondary syphilis are always low, e.g. 1 : 20 to 1 : 320. However, these titers seem to be significant when heterophilic antibodies are elimininated by absorption onto non-sensitized erythrocytes. Furthermore, all the BFP gave negative results.

False positive reactions to the Reiter protein complement-fixation (RPCF) test.

False positive reactions to the Reiter protein complement-fixation (RPCF) test.

Early Syphilis: Immunoglobulins Reactive in Immunofluorescence and other Serologic Tests

Summary Sera and serum fractions from 41 patients with darkfield-positive untreated primary and secondary syphilis were studied to reveal the immunoglobulins that are reactive in immunogluorescence and other serologic tests for syphilis. A monospecific indirect fluorescent antibody (IFA) assay revealed that IgG is the major immunoglobulin class reactive with Treponema pallidum, and that smaller amounts of IgM and IgA antibodies may also react. IgG is the principal immunoglobulin operative in the fluorescent treponemal antibody-absorption (FTA-ABS) test, and in its predecessor tests, the fluorescent treponemal antibody 1:5 (FTA-1:5), and the fluorescent treponemal antibody-200 (FTA-200). “Sorbent,” or a sonicate of Reiter treponemes, which is used in the FTA-ABS test to distinguish syphilitic from nonsyphilitic sera, operationally acts mainly to prevent “natural” IgG antibodies in nonsyphilitic serum from reacting with T. pallidum. It does not prevent syphilitic IgG antibodies from reacting, and apparently it enhances their role relative to IgM and IgA antibodies. Reactivity in the Treponema pallidum immobilization (TPI) test and the Reiter protein complement-fixation (RPCF) test was found mainly in the 7 S fractions of sera as compared to the 19 S fractions. Absorbing a syphilitic serum with T. pallidum did not produce a measurable decrease in total levels of IgG, IgM, or IgA. Antilipoidal antibodies detectable in the VDRL slide test were found in both the 19 S and 7 S fractions of sera; the antibodies associated with primary syphilis were not confined to the IgM class. In the course of these studies, 19 S and 3.5 S IgG-related substances were noted to possess anti-T. pallidum reactivity, and IgA in the 7 S fraction of serum was observed to be functionally silent in contrast to IgA of higher weight.

Advantage of a routine Reiter protein complement-fixation test in the serodiagnosis of syphilis in pregnancy.

One hundred and twenty-seven (0.3%) reactive serological tests for syphilis were obtained by routine examination of sera from 35,912 antenatal patients.Forty-eight (38%) of these reactive sera were associated with reactive fluorescent treponemal antibody and Treponema pallidum immobilization results and these patients were therefore considered to show serological evidence of treponemal infection. Sera from 13 of these 48 patients (27%) gave reactive Reiter protein complement-fixation tests in the absence of detected ;reagin' antibody. Seventy-nine patients showed non-specific biological false positive reactions to the routine tests. Following clinical assessment of these serologically reactive patients, approximately one in 100 immigrant and one in 2,500 non-immigrant pregnant women examined in this series were treated with penicillin; this represented just over half the number of patients with serological evidence of a treponemal infection, a ratio which was similar for immigrant and native born women. Yaws was the likely treponemal infection in the majority of infected immigrants. The significance of the serological results and the advantages of the Reiter protein complement-fixation test are discussed.

Treponema pallidum Reiter株の抗原構造分析と梅毒血清反応における意義

A series of studies has been conducted about the antigenic structure of the Reiter Strain of Treponema pallidum using a refined Reiter protein antigen purified by lower phase solvent adapting the Agar Gel Diffusion test and Complement Fixation test. Through these studies, the existence of the common antigen between the Reiter strain and pathogenic treponema (Nichols strain) was observed and furthermore this chemical component was acknowledged as the protein. However, the removal of appearance of the nonspecific reaction was nearly impossible at the RPCF test using this purified Reiter antigen.This should be considered, as the author has refered previously, to be caused by the common antigen of the saprophytic treponema existing in the healthy human.On the other hand, the positive reaction was observed at the serologic test for the human serum from the syphilitic individual absorbed by the Reiter strain using the pathogenic treponema (Nichols strain) as an antigen, and from this results, it was found the Reiter strain has only the common antigen which is exist among treponemes except own specific antigen, and the pathogenic treponema has the specific antigen other than the Reiter strain.From the test aforementioned the usage of the Reiter strain as an antigen at Complement Fixation test is not appopriate, and rather it should be used as an absorption antigen for nonspecific reaction caused by the common antigen among treponemes at the test involving the Fluorescent Treponemal Antibody test where the pathogenic treponema is being used as an antigen.

Cryoglobulinemia Presenting as "Factitial Ulceration"

A case of "essential" cryoglobulinemia was initially misdiagnosed as factitial ulceration. The abnormal protein, not demonstrable on routine electrophoresis, was characterized as a macrocryoglobulin. A biologic false-positive reaction occurred with the venereal disease research laboratory test for syphilis (VDRL), and the Kolmer and fluorescent treponemal antibody (FTA-200) tests, but the Reiter protein complement fixation test (RPCF), fluorescent treponemal antibody test with serum absorbed by Reiter protein sonicate (FTA-abs), and<i>Treponema pallidum</i>immobilization test (TPI) were nonreactive. Pitfalls in diagnosis of cryoglobulinemia are enumerated. Clinical criteria for factitial ulceration are applied.

The Reiter protein complement-fixation test using the Auto-Analyzer.

A method for the Reiter protein complement-fixation test using the Techncon AutoAnalyzer is described. Two hundred and fifty sera were tested, when six false-positive and nine false-negative results were obtained when compared with results obtained by the manual method. Deep freezing the sera before testing on the AutoAnalyzer increased the sensitivity so that in a further 250 sera tested no false-negative and two false-positive results were obtained when compared with the results obtained at the Venereal Diseases Reference Laboratory, London.

Performance and use of the Reiter protein complement-fixation (RPCF) test.

Performance and use of the Reiter protein complement-fixation (RPCF) test.

Immunity in experimental syphilis. IV. Serological reactivity of antigens extracted from gamm-irradiated Treponema pallidum and Treponema reiteri.

Miller, James N. (University of California School of Medicine, Los Angeles), J. H. De Bruijn, and J. H. Bekker. Immunity in experimental syphilis. IV. Serological reactivity of antigens extracted from gamma-irradiated Treponema pallidum and Treponema reiteri. J. Bacteriol. 91:583-587. 1966.-Ultrasonic lysate preparations extracted from virulent Treponema pallidum, Nichols strain, suspensions exposed to 652,800 R of gamma-irradiation exhibited a loss in the serological reactivity of their heat-labile antigens; the heat-stable components of both the lysate and residue antigens were unaffected. The activity of heat-stable, cardiolipin T. pallidum complement-fixing antigen obtained from similarly irradiated organisms was also unaltered. gamma-Irradiation of the cultivable Treponema reiteri with dosages as high as 6,500,000 R failed to alter serologically either the heat-labile or heat-stable component of its lipopolysaccharide-protein (Reiter protein) antigen. The reactivity of the lipopolysaccharide portion of the Reiter protein complex with an antiserum to T. pallidum Nichols indicates previously unsuspected antigenic differences between the rabbit-adapted Nichols strain of the organism and so-called "wild" human strains of T. pallidum in which this antigen is generally absent.

Treponemal antigens as related to identification and syphilis serology.

Summary1. The antigens of intact T.p., the R.t., T.m. and the free-living mud organism, T.z., are compared. 2. An antigen appearing to be identical to the Reiter Protein antigen was found to be a common or shared component in all of the treponemes studied. 3. By means of absorption or blocking procedures and employing FA methods, all treponemes could be identified. 4. It is suggested that FA methods can play an important part in future taxonomic studies of treponemes and that the FTA test can be modified so as to increase both sensitivity and specificity.

Evaluation of fluorescent treponemal antibody (RFTA and FTA II) and other tests (RPCF and TPI) for syphilis.

SummaryFluorescent treponemal antibody tests (RFTA and FTA II) using hypochlorite-treated Reiter and Nichols strain treponemes, respectively, were compared with each other and with a Reiter protein complement-fixation (RPCF) test in a study of sera previously subjected to the Treponema pallidum immobilization (TPI) test. The fluorescent antibody (FA) tests were closely comparable in sensitivity and specificity for syphilis. The RPCF test was slightly more sensitive than either FA test, but yielded somewhat more “non-specific” reactions. It appeared that either FA test might serve as well as the RPCF test as a screening procedure in differentiating persons with latent syphilis from those whose sera react “non-specifically” with cardiolipin antigens. Considerations favoring selection of an FA test are simplicity of performance and the need for fewer reagents. Moreover, the RFTA test substitutes an organism that can be cultivated simply in vitro for one requiring maintenance in infected rabbits.

Long-term serologic studies with treponemal procedures.

The purpose of this paper is to document our experience with treponemal testing since 1955 in a series of 1,064 clinically evaluated patients, and to discuss the patterns of seroreactivity after therapy in the various stages of syphilitic infection. We have been fortunate in being able to serologically evaluate certain cases for as long as 25 to 30 years after treatment. The tests used in this study consist of specific treponemal procedures as well as a battery of standard serologic tests for syphilis. The treponemal procedures employed were the Treponema pallidum complement-fixation 1 (TPCF) test, the Reiter protein complement-fixation 2 (RPCF) test, and the Treponema pallidum immobilization (TPI) test. All of the serums studied were subjected to the TPCF and RPCF tests in our laboratory. A small number of serums were tested with the TPI test which was performed by Dr. James N. Miller of the University of California Medical