ON THE PRESENCE IN SYPHILITIC SERUM OF ANTIBODIES TO SPIROCHETES,THEIR RELATION TO SO CALLED WASSERMANN REAGIN,AND THEIR SIGNIFICANCE FOR THE SERODIAGNOSIS OF SYPHILIS

1. In confirmation of Gaehtgens, syphilitic human sera give positive complement fixation with cultures of so called T. pallidum (Reiter strain). Syphilitic rabbit sera are equally reactive. Syphilitic human and rabbit sera agglutinate these cultures, often in high titre (Beck). 2. Normal rabbit sera react weakly with the culture to give both agglutination and complement fixation in low titre. Normal human sera, despite the fact that they contain agglutinins in low titre, fail to fix complement with the Reiter strain of cultured spirochetes. Confirming Gaehtgens, the latter reaction is therefore of practical utility for the serum diagnosis of syphilis. 3. When syphilitic serum is heated at 63°C., there is no demonstrable difference in the thermolability of the antibody to spirochetes, and of the reagin which determines the Wassermann and flocculation tests. 4. (a) The absorption of syphilitic serum by spirochetal suspensions removes all reactivity, not only for the spirochetes, but for tissue lipoids (alcoholic beef heart extract) as well; the sera become Wassermann- and flocculation-negative. (b) Absorption of syphilitic serum with tissue lipoids renders the Wassermann and flocculation tests negative, but does not demonstrably change the reactivity of the serum with spirochetes. (c) Rabbits immunized to beef heart lipoid develop spirochetal agglutinins and complement-fixing antibodies (Reiter strain) in high titre. 5. It is concluded that these cultured spirochetes contain antigenic material serologically related to a substance present in mammalian tissue, as well as other antigenic factors not present in such extracts, but equally reactive with syphilitic serum. 6. These findings support the thesis that the primary serologic change in syphilis is the development of antibodies to T. pallidum. The Wassermann and flocculation tests would be explained on the basis that the tissue extracts used as "antigen" in these tests contain one or more substances serologically related to antigenic components of T. pallidum. Similarly, the cultured Reiter strain of spirochete is apparently sufficiently close serologically to T. pallidum to be agglutinated by and to give complement fixation with the antibodies to T. pallidum present in syphilitic serum. 7. Since suspensions of cultured spirochetes contain antigenic factors which react specifically with syphilitic serum, some of which are not present in ordinary Wassermann and flocculation "antigens," they may prove even more valuable than those tissue extracts in the serodiagnosis of syphilis.     

STUDIES ON TREPONEMA PALLIDUM AND SYPHILIS : IV. THE DIFFERENCE IN BEHAVIOR IN IMMUNE SERUM BETWEEN CULTIVATED NON-VIRULENT TREPONEMA PALLIDUM AND VIRULENT TREPONEMATA FROM LESIONS.

Although antibodies can be produced by the immunization of animals with cultivated Treponema pallidum, and although these antibodies exert specific agglutinative and treponemicidal action upon the culture organisms, they possess, at least in the concentration so far obtained by us in rabbits and sheep, practically no action for virulent treponemata obtained directly from lesions. There seems to be in the infected body an inability to exert a purely serum action upon the virulent treponemata, a condition of affairs which may well lead to a lack of antigen absorption on the part of the body and a consequent failure to produce serum antibodies. We do not think that this should in any way discourage our further investigation of the protective action of antibodies produced with culture pallida. On the one hand, the slight occasional agglutination and the lower proportion of takes with the concentrated serum in the last experiment at least indicate the possibility that we have been working with sera that are not sufficiently powerful and that just as with work with the pneumococcus and other highly invasive organisms, a serum of considerable antibody contents must be used before results can be expected. Again, the destruction of treponemata and the healing of lesions which undoubtedly takes place in rabbits, sometimes with surprising speed, may be a cellular destruction, and by injecting the sera either locally or intravenously and giving them time to be absorbed by the cells before injecting virulent material, better results may be obtained. This direction of research as well as further studies on the antagonistic cellular processes against the pallida, the immunization of animals with killed virulent organisms, and the antibodies in rabbits and human beings during the course of infection and after recovery are being investigated, and we hope to be able to report upon them in the near future.     

STUDIES IN EXPERIMENTAL SYPHILIS : VI. ON VARIATIONS IN THE RESPONSE OF TREATED RABBITS TO REINOCULATION; AND ON CRYPTOGENETIC REINFECTION WITH SYPHILIS.

Syphilitic rabbits inoculated intratesticularly and treated with arsphenamine before the 69th day of the disease, when reinoculated with the same strain of treponemata and in a manner identical with that of the first inoculation) are capable of responding to the infection in at least five different ways. In addition to exhibiting a local lesion at the site of reinoculation, accompanied by dissemination of the virus, they may show no local lesion at all but present evidence of dissemination of the virus together with the development of a positive Wassermann reaction. In some instances the virus may be recovered from the reinoculation site although no local lesion is produced there and no dissemination of the virus can be shown to take place. An occasional animal treated before the 69th day of the disease remains completely refractory to a second infection. When treatment is postponed to 6 months or more after the original inoculation, reinfection carried out by intratesticular injection is almost always impossible and such animals appear to be entirely refractory. However, if rabbits treated late are reinoculated with the homologous strain by depositing the virus upon a granulating wound on the back, successful reinfections can be accomplished in at least 50 per cent of the test animals. The resistance which develops in rabbits in the course of syphilitic infection is, then, not absolute but relative. It is pointed out that these results cast discredit upon the validity of the reinoculation method as a test of cure in syphilitic infection. It is also suggested upon the basis of these experiments, that the subsequent occurrence of a positive Wassermann reaction in patients with early syphilis in whom the Wassermann reaction has become negative under treatment may not always represent a relapse in the the disease but possibly in some instances a new infection without clinical signs.     

Monoclonal antibody with hemagglutination, immobilization, and neutralization activities defines an immunodominant, 47,000 mol wt, surface-exposed immunogen of Treponema pallidum (Nichols)

Radioimmunoprecipitation (RIP) analyses performed on 125I-surface-labeled Treponema pallidum cells using various immune sera revealed the presence of six major surface antigens (immunogens) with apparent molecular weights of 47 K, 36 K, 34 K, 32 K, 29 K, and 13 K. Among these, the 47 K surface antigen was most abundant. Radioimmunoprecipitation assays using 125I-labeled T. phagedenis biotype Reiter or immunoblot analyses using the same strain, failed to reveal the presence of the 47 K mol wt antigen in the representative nonpathogenic treponeme. Preabsorption of anti-T. pallidum immune rabbit serum (IRS) with the Reiter organism did not remove anti-T. pallidum antibodies from immune serum that reacted with the 47 K mol wt immunogen or other immunogens of T. pallidum present in the characteristic antigenic profile. Monoclonal antibodies (mAb) directed specifically against the 47 K mol wt immunogen of T. pallidum also failed to react with an analogous 47 K mol wt component in Treponema phagedenis biotype Reiter, further suggesting the unique presence of this antigen in pathogenic treponemes. The presence of the 47 K mol wt surface immunogen in pathogenic treponemes other than T. pallidum subspecies pallidum was also observed (43). Anti-47 K immunogen mAb was nonreactive against rabbit IgG or IgM. mAb directed specifically against the 47 K mol wt immunogen of T. pallidum was examined for strategic functional activities. It was found to be reactive in the microhemagglutination assay for T. pallidum antibodies, the T. pallidum immobilization test, and was found to be capable of significant blockage of attachment of virulent T. pallidum to host cells in tissue culture. Additional significant biological activity for the anti-47 K mol wt immunogen mAb was revealed through results of the in vitro-in vivo neutralization test of Bishop and Miller, in which a 99% or 100% neutralizing activity was demonstrated. The combined data of this study suggest that the 47 K mol wt immunogen of T. pallidum represents an abundant, immunodominant, surface-exposed immunogen possessing potential biological importance in the pathogenesis and immunology of T. pallidum infection. These studies serve to establish the first functionally defined immunogen for T. pallidum, which may represent the major immunogen of the organism.     

STUDIES IN THE SEROLOGY OF SYPHILIS : VI. THE INDUCTION OF ANTIBODIES TO TISSUE LIPOIDS (A POSITIVE WASSERMANN REACTION) IN NORMAL RABBITS

More than one-half of normal rabbits contain complement-fixing or precipitating antibodies against Wassermann antigens (the alcohol-soluble lipoids of beef, rabbit, and human hearts) by a sufficiently sensitive technique. Normal human sera tested by the same technique are uniformly negative. The intravenous injection of colloidal suspensions of beef and human heart lipoids into rabbits occasionally causes a significant increase in this normal Wassermann (antilipoid) titre. This may indicate a certain degree of antibody response to the lipoids as such; it may be due to the presence in such extracts of traces of foreign protein, which would activate the lipoid haptene into a complete antigen; or it may be a non-specific increase in a normal antibody, not due to a specific antigenic stimulus. Confirming the results of Sachs, Klopstock, and Weil, the addition of normal foreign (human) serum to rabbit, beef, and human heart lipoids makes them antigenic for rabbits. The intravenous injection of such lipoid-serum mixtures usually causes a significant increase in the titre of the complement-fixing and precipitating antibody against tissue lipoids. The precipitate which forms upon the addition of tissue lipoids to human syphilitic serum is by far the most efficient antigen for the production, in rabbits, of antibodies to tissue lipoids which we have as yet encountered. Rabbits injected intravenously with such a precipitate regularly develop a Wassermann titre which is many times higher than either the titre observed in human syphilis, or that induced by the injection of a normal serum-lipoid mixture. The very marked antigenic property of the precipitate as compared with that of a normal serum-lipoid mixture is considered to be due to the fact that it contains a foreign protein firmly bound to the lipoid particles, namely, the human reagin-globulin with which they have combined. This interpretation is supported by the observations (1) that heating at 100°C., which does not affect the lipoid constituent of the precipitate, destroys its antigenic power for rabbits, and (2) that a similar precipitate derived from Wassermann positive rabbit serum instead of syphilitic human serum, and therefore containing tissue lipoid in combination with homologous (rabbit) protein, is completely non-antigenic for rabbits.     

A STUDY OF THE RELATION OF TREPONEMA PALLIDUM TO LYMPHOID TISSUES IN EXPERIMENTAL SYPHILIS

A widespread dissemination of Treponema pallidum from a local focus of inoculation in the rabbit constantly occurs by way of the lymphatics. Spirochetes were regularly recovered from the satellite lymph nodes by animal inoculation after scrotal inoculation; they were present as early as 2 days, when no specific primary reaction was detected, and at later periods of from 5 to 61 days after inoculation. Other superficial nodes at remote sites such as the popliteals and with no syphilitic lesions in the drainage area have also been shown to harbor active organisms. Although spirochetes were found in relatively few of the lymph node emulsions, the orchitis resulting from their injection was of a rapidly progressive type with an incubation period but slightly longer than that produced by a testicular or skin nodule emulsion rich in spirochetes. It has further been shown that a syphilitic infection is sufficiently established in the rabbit body within 48 hours after scrotal inoculation so that the primary lesion is no longer essential for its maintenance. Active treponemata survive in the popliteal lymph nodes for long periods of time and have been regularly recovered from them in cases of true latency. The lymph nodes, therefore, function as reservoirs of the organisms. The ability to recover the spirochetes from lymphoid tissue through successive generations is seen in the serial passage of lymph node emulsion to testicle during an 18 months period. The persistence of spirochetes in lymphoid tissue irrespective of the presence or absence of syphilitic lesions is a characteristic and fundamental feature of syphilis of the rabbit. The existence of infection, therefore, may be demonstrated at any time by the recovery of spirochetes from the popliteal lymph nodes by animal inoculation. This fact is of great practical importance in the therapy of the infection and may be profitably utilized in determining the ultimate effect of a therapeutic agent. These experiments demonstrate that the disease is not confined to the site of local inoculation but that lymphogenous dissemination of treponemata regularly takes place, and that during the course of this process organisms become localized in the lymph nodes and exist there indefinitely irrespective of the occurrence of manifestations of disease. The intimate relation of Treponema pallidum to lymphoid tissue is an essential concept of syphilis of the rabbit, and from this point of view, the infection is primarily one of lymphoid tissue.     

STUDIES IN THE SEROLOGY OF SYPHILIS : II. THE PHYSICAL BASIS OF THE WASSERMANN REACTION

The substance in syphilitic serum which is responsible for the Wassermann reaction, like that which determines the diagnostic flocculation tests, is associated with the globulin fraction of serum. Every positive Wassermann is accompanied by microscopic (or submicroscopic) aggregation, which is not an essential feature of the reaction; conversely, after every positive flocculation test, the washed precipitate will fix complement. An excess of antigen removes both flocculating and complement-fixing substances completely (>95 per cent). Heating the lipoid-reagin precipitate to 100° for 1 minute destroys the sensitizing film of reagin globulin; the avidity for complement disappears simultaneously. Both the flocculating and complement-fixing properties of syphilitic serum are therefore determined by the same substance, a specifically altered fraction of the serum globulin, reagin. The Wassermann reaction is thus entirely analogous to complement fixation by any antigen-antibody complex. The same film of denatured serum globulin which sensitizes the antigen particles, whether red cells, bacteria, protein, or colloidal lipoid particles, to discharge and aggregation by electrolytes, also endows them with an avidity for complement. The pathogenesis of reagin will be discussed in a forthcoming paper.     

The significance of Karl Landsteiner's works for syphilis research

On January 7th 1905, more than five months before the detection of T. pallidum, Karl Landsteiner began his work on syphilis research together with notable members of the Viennese School of Medicine, namely Ernest Finger, Rudolf Müller, Viktor Mucha, Otto P?tzl and others. Extensive animal experiments led to the formulation of the Finger-Landsteiner Law and provided the basic facts for the Jadasson-Lewandowsky Law. Attempts of active or passive immunization were unsuccessful and, indeed, were still a failure in 1990 after implementation of the latest tools of modern research, including gene technology. Dark-field microscopy was introduced for the detection of T. pallidum by Landsteiner and Mucha. These authors noted that serum of syphilitic patients inhibited the movements of T. pallidum and, thus, observed the basic principle underlying the T. pallidum immobilization test (= TPI = Nelson-Mayer test). Finally, Landsteiner, Müller and P?tzl discovered that it was not an antibody specific to T. pallidum that reacted in the Wassermann reaction, but "autotoxic" substances, which they called reagines. During the 1970's and 1980's it was discovered that these reagines are autoantibodies directed against parts of the inner envelope of the mitochondria.     

Crosslinking CD4 by human immunodeficiency virus gp120 primes T cells for activation-induced apoptosis

During human immunodeficiency virus (HIV) infection there is a profound and selective decrease in the CD4+ population of T lymphocytes. The mechanism of this depletion is not understood, as only a small fraction of all CD4+ cells appear to be productively infected with HIV-1 in seropositive individuals. In the present study, crosslinking of bound gp120 on human CD4+ T cells followed by signaling through the T cell receptor for antigen was found to result in activation-dependent cell death by a form of cell suicide termed apoptosis, or programmed cell death. The data indicate that even picomolar concentrations of gp120 prime T cells for activation-induced cell death, suggesting a mechanism for CD4+ T cell depletion in acquired immune deficiency syndrome (AIDS), particularly in the face of concurrent infection and antigenic challenge with other organisms. These results also provide an explanation for the enhancement of infection by certain antibodies against HIV, and for the paradox that HIV appears to cause AIDS after the onset of antiviral immunity.     

COMPLEMENT DEVIATION IN SCARLET FEVER WITH COMPARATIVE STUDIES OF THE WASSERMANN AND NOGUCHI SYSTEMS

The Wassermann reaction in scarlet fever per se is uniformly negative. The antigen of scarlet fever liver yields practically the same results as that of luetic liver, and both fail to deviate complement with scarlet fever antibodies. The Noguchi reaction in scarlet fever is practically negative. Sixteen cases, or 6.4 per cent. of 250 cases, were positive when active serum was used; with inactivated serum, but eleven, or 4.4 per cent., remained positive. Five of these eleven cases were also positive with the Wassermann system. In other words, sixteen cases, or 6.4 per cent., were positive according to the Noguchi system with active or inactivated serum or both, whereas with the Wassermann system only 2 per cent. were positive. The presence of anti-sheep hemolysin normally in human serum is one of the main disturbing factors in the Wassermann system; for this reason, complement and hemolysin (made by immunization of rabbits) require careful titration. A positive Wassermann reaction usually indicates the presence of syphilitic antibodies, and a negative Noguchi reaction, their absence, and both systems should be used in the examination of all cases.     

EPSTEIN-BARR VIRUS : HETEROPHILE RESPONSES IN SQUIRREL MONKEYS INOCULATED WITH VIRUS-TRANSFORMED AUTOLOGOUS LEUKOCYTES

Epstein-Barr virus (EBV)-transformed autologous lymphoblasts were repeatedly inoculated into three squirrel monkeys. Each animal developed the heterophile antibodies of infectious mononucleosis and EBV-specific antibodies. After serologic responses had disappeared or markedly declined, the animals were challenged with either whole cells, cell filtrate, or cell ghosts. Animals challenged with living cells and cell ghosts developed agglutinin responses; the recipient of filtrate did not. The results suggest that EBV induces the appearance of the infectious mononucleosis heterophile antigen on the transformed cell membrane.     

Fibronectin mediates Treponema pallidum cytadherence through recognition of fibronectin cell-binding domain

The specificity of the interaction between Treponema pallidum and fibronectin was demonstrated. Treatment of host cells with only antifibronectin sera and not anticollagen or antilaminin sera, inhibited treponemal cytadsorption. Incubation of fibronectin-coated coverslips with monoclonal antibody to the cell-binding domain of fibronectin reduced treponemal attachment to the same extent as antifibronectin serum. Both iodinated fibronectin and iodinated cell-binding domain bound to T. pallidum in a saturable manner. Specificity of the T. pallidum association with the cell-binding domain was the most effective inhibitor of the binding of either radioiodinated cell-binding domain or fibronectin to T. pallidum. Scatchard analysis gave Kd on the order of 10(-7) M for both cell-binding domain and fibronectin binding to T. pallidum, consistent with the high affinity interaction of these organisms with host cell surfaces. Finally, the same level of attachment of treponemes was achieved on coverslips coated with cell-binding domain as that observed for organisms incubated with fibronectin, indicating that the cell-binding domain polypeptide is functionally identical to fibronectin in mediating T. pallidum adherence.     

EXPERIMENTS ON THE PRODUCTION OF WASSERMANN REAGINS BY MEANS OF TRYPANOSOMES

Since it is known that positive Wassermann reactions prevail in trypanosomiasis of rabbits, similar to those in syphilis, trypanosomes were used for an inquiry into the cause of this reaction. Injections with dead trypanosomes into rabbits proved that these microbes are highly active antigens and suffice in themselves to produce strongly positive Wassermann sera, in analogy to the findings reported by F. Klopstock with Spirochæta pallida. Although a number of questions require further study, yet it seems likely that this antigenic activity of the microbes or their products plays a part in the production of the Wassermann reagins in infections with spirochetes and trypanosomes.     

ANTIBODY FORMATION AGAINST TREPONEMA PALLIDUM—AGGLUTINATION

It has been shown by our experiments that the serum of rabbits treated with emulsions of Treponema pallidum contains agglutinating substances. Normal rabbit serum also possesses agglutinating power for this organism, but, as in the case of normal bacterial agglutinins, to an extent very much inferior to that possessed by the sera of immunized animals. Normal human sera will agglutinate similar pallidum emulsions, as will the sera of certain syphilitic patients with positive Wassermann reactions. Whether or not there is a quantitative difference of diagnostic value between the sera of normal human beings and those of syphilitics remains to be seen. The sera of rabbits immunized with strain A agglutinate Noguchi''s strain 9 in dilutions as high as 1 to 500. We regard as the most important result of these experiments the demonstration of definite antibodies in the circulation of animals treated with dead emulsions of Treponema pallidum. Since it is our belief that the agglutinating effect is due to an antibody essentially the same as that which produces bactericidal, precipitating, and opsonic effects, i. e., that there is probably one type of antibody only, we believe that the demonstration of agglutinins establishes the fact that in syphilis as in bacterial diseases the host responds by the formation of antibodies or sensitizers specific for the treponema. Spirocheticidal experiments with these sera, both in vitro and in vivo, are in progress.     

Evaluation of Sch 29482 in experimental syphilis and comparison with penicillin G benzathine in disseminated disease and localized infection

The present study was designed to assess the in vivo activity of Sch 29482, a new penem antibiotic, against disseminated and localized Treponema pallidum infections in rabbits. Animals were inoculated either intravenously or intradermally. Randomized groups then received 25 or 50 mg of Sch 29482 per kilogram of body weight twice a day for 7 days, two weekly injections of 200,000 U of penicillin G benzathine for comparative purposes, or no antibiotic therapy. In both infection models, striking differences were noted between the untreated control rabbits and rabbits receiving penicillin G benzathine or high-dose Sch 29482. Intravenously infected rabbits did not develop disseminated lesions or orchitis, and chancres produced by intradermal infection regressed and healed rapidly after both treatment regimens. Infectivity studies also suggested that high-dose Sch 29482 and penicillin G benzathine were effective since the testes and lymph nodes of treated animals were free of infectious organisms. Treatment of animals with the lower dose of Sch 29482 represented borderline or suboptimal therapy, with a failure rate of one in four for each infection model.     

经修饰的重组梅毒螺旋体膜蛋白及其应用

目的　寻找高灵敏度、高特异性、检测结果定量、简便易行的梅毒螺旋体抗体酶联免疫检测方法。　方法　采用重组技术 ,在现有已公布的梅毒螺旋体基因组中筛选出公认抗原性最强的三个基因片段 Tpp1 5、Tpp1 7、Tpp47进行克隆。为了获得水溶性高、特异性强、可标记性好的重组抗原蛋白 ,有针对性地设计引物以便对表达的梅毒螺旋体膜蛋白进行修饰和改造 ,同时在收获高效表达的目的蛋白后采取了一系列针对性切割和高特异性纯化。以纯化的重组梅毒螺旋体膜蛋白 (r-KTp1 5 ,r-KTp1 7及 r-KTp47)为抗原 ,建立了诊断梅毒螺旋体抗体的双抗原夹心酶联检测方法 (DAGS ELISA)。　结果　在大肠杆菌中获得了重组梅毒螺旋体膜蛋白的高效表达 ,经修饰纯化之后的目标蛋白水溶性提高 ,特异性增强 ,可标记性优于常规不修饰的重组蛋白。应用该三种蛋白为抗原建立的双抗原夹心法血清学诊断试剂盒共检测各期梅毒阳性患者 2 1 0例 ,灵敏度达 98.6% (2 0 7/2 1 0 ) ,检测正常献血者 1 3 2 7例 ,特异性达 1 0 0 .0 % (1 3 2 7/1 3 2 7) ,优于常规梅毒血清学初筛诊断方法 RPR及 TRUST法 ,与 TPHA法的符合率为 1 0 0 .0 %。　结论　以双抗原夹心法酶联免疫诊断试剂盒在梅毒血清学初筛实验室进行梅毒螺旋体抗体的筛查 ,在灵敏度、特异     

ACUTE VASCULAR LESIONS IN MICE FOLLOWING INJECTIONS OF PNEUMOCOCCI

Mice dying several days after injections of pneumococci, both living and dead, frequently show at autopsy large intrathoracic hemorrhages. The histological study of the thoracic organs indicates that there occurs in each case a sharply circumscribed, acute degeneration of the wall of some large vessel, usually the ascending aorta or one of the pulmonary arteries. This degenerated portion is torn out by the pressure of the blood with almost complete disappearance of the vessel wall, leading to a gross hemorrhage. A similar change is occasionally found in the walls of the veins which contain cardiac instead of smooth muscle. We have found this lesion only in mice which had been recently inoculated with pneumococci. Negative cultures at autopsy, the lack of inflammatory reaction, and the occurrence of the conditions after injection of dead pneumococci suggest the cause to be a toxic degeneration of the vessel wall brought about by the poisons of the injected organisms.     

Electroimmunodiffusion reaction in the diagnosis of syphilis (author's transl)]

A group of 800 human sera, obtained from syphilitic and nonsyphilitic individuals were tested in the counterimmunoelectrophoresis (CIE) technique with Reiter protein antigen. The sera were assayed for comparison in the following treponemal and nontreponemal tests: standard serologic tests for syphilis with cardiolipin; complement fixation (Kolmer 1/5 vol) with Reiter protein antigen (RPCF); T. pallidium immobilization (TPI) test; haemoagglutination test for T. pallidum antibodies (TPHA). The sensitivity and specificity of CIE test, its simplicity, the low cost, the possibility of practical application as a routine test for syphilis serology were discussed.     

DISTINCTIVE CHARACTERISTICS OF INFECTIONS PRODUCED BY TREPONEMA PERTENUE IN THE RABBIT

From a study of rabbits inoculated intratesticularly with two strains of treponemata derived from patients suffering from clinical yaws, it was found that a characteristic feature of the reaction to the infection was a well marked periorchitis of a granular or finely nodular type with or without a diffuse involvement of the tunic. While lesions of the testicular parenchyma also occurred they were relatively inconspicuous and consisted either of a minor diffuse orchitis which was usually followed by atrophy and fibrosis of the organ, or of small nodules; or there might be a combination of diffuse and nodular lesions. The granular periorchitis could be recognized clinically almost as soon as any change could be detected in the testicle, that is, about 3 weeks after inoculation; in the following month the covering of the testicle became studded with numerous tiny indurated nodules. Subsequently regression and healing took place and in the majority of animals no lesions were found 3 months after inoculation. In some animals residual lesions persisted for as long as 6 months. The granular periorchitis was a practically constant feature of the infection and was unlike any lesion of the tunic observed in experimental syphilis of the rabbit. Treponemata were numerous in the lesions of the tunic and somewhat less so in the testicle itself. Dissemination of organisms to the uninoculated testicle and to the inguinal lymph nodes was demonstrated by animal inoculation although the clinical signs of a metastatic orchitis and periorchitis were slight and a lymph adenitis was an inconstant feature of the infection. Generalized lesions in remote parts of the body, similar to those occurring in experimental syphilis of the rabbit, were not observed.     

FURTHER OBSERVATIONS ON THE RELATION OF THE EYE TO IMMUNITY IN EXPERIMENTAL SYPHILIS : III. THE INFLUENCE OF A NON-SPECIFIC INFLAMMATORY REACTION IN THE CORNEA ON THE DEVELOPMENT OF IMMUNITY IN THAT TISSUE AFTER INTRATESTICULAR INOCULATION

Two experiments are reported in which rabbits originally inoculated with syphilis and treated late in the course of the disease (174th to 210th day) were reinoculated subsequently in both corneas with the homologous strain of syphilitic virus. In each animal one cornea was inoculated with dead tubercle bacilli prior to reinoculation with the syphilitic virus. This procedure was carried out in order to bring about a non-specific inflammatory reaction with resultant vascularization, the intention being to find out if such vascularization would render the cornea more resistant to inoculation with the homologous strain of syphilitic virus. The results of both experiments were similar and while they were not conclusive, they indicated that there was a tendency for corneas which had been injected with dead tubercle bacilli to be more refractory to a subsequent inoculation with homologous syphilitic virus than the corneas of the same animals that had not been injected with dead tubercle bacilli. This tendency may be interpreted as suggestive evidence for the view that in the syphilitic rabbit there develop circulating antibodies toward the homologous strain of T. pallidum.