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Humoral response in Treponema pallidum-infected guinea pigs: I. Antibody specificity.

Young male inbred strain 2 guinea pigs were infected intradermally with 8 X 10(7) Treponema pallidum extracted from a rabbit orchitis, and 5 months later reinfected with 10(7) T. pallidum. Ninety percent of the animals developed symptomatic lesions after initial infection but none on challenge. Immunoblotting of sera obtained at intervals after infection or reinfection showed antibodies against T. pallidum antigen (TP), nonpathogenic treponemes--T. phagedenis biotype Reiter (TR), T. refringens strain Noguchi (TN), and T. vincentii (TV)--as well as normal rabbit serum (NRS) and normal rabbit testes extract (NRT). Antibodies reacting with TP were detected as early as 17 days (five polypeptides) and steadily rose (at 3 months 17 polypeptides were seen). Cross-reacting antibodies to TR, TN, TV, or rabbit proteins decreased within 3 to 5 months. After reinfection, the antibodies to NRS increased more sharply than the anti-treponemal antibodies. Adsorption with TR and NRS of sera obtained after infection or reinfection produced a reduction of antibodies to TP by 75-87%.

Animals↗

Kinetics of pathogen-specific humoral response in Treponema pallidum-infected young and old inbred strain 2 guinea pigs.

The kinetics of the humoral response to pathogen-specific polypeptides was examined in Treponema pallidum-infected young (3-5 months old) and old (10-20 months old) inbred strain-2 guinea pigs. Sera collected before and at various times after infection were pooled and examined by immunoblotting and two serologic tests (ELISA and FTA) before and after sequential adsorption with CNBr-activated sepharose coupled to normal rabbit proteins and antigens from five nonpathogenic treponemal species. Prior to adsorption the kinetics of the humoral response to T. pallidum antigens did not seem to differ significantly between the two groups. After adsorption, however, a delay in the appearance of detectable antibodies and a milder response to various pathogen-specific polypeptides was observed in the older group. After adsorption, a sharp drop in ELISA-TP, ELISA-TR and FTA titres occurred in both groups. Six months post-infection, between 9 and 10 pathogen-specific polypeptides (97, 57, 47, 45, 43, 39, 37, 33, 17 and 15 kD) were recognized by both groups. The effect of age and levels of natural treponemal antibodies on the clinical symptoms of primary lesions and humoral response to pathogen-specific polypeptides is discussed.

Aging↗

Pathogen-specific humoral response in Treponema pallidum-infected humans, rabbits, and guinea pigs.

Molecular analysis of the humoral response to pathogen-specific polypeptides was done using sera from patients at different stages of syphilis and sera from Treponema pallidum-infected rabbits and guinea pigs collected at various times after infection. The sera were examined by ELISA, fluorescent treponemal antibody absorption test (FTA-ABS), and immunoblot before and after sequential adsorption with cyanogen bromide-activated Sepharose 4B coupled individually to five sonicated nonpathogenic treponemes. Guinea pig antisera were also adsorbed with rabbit proteins. After adsorption, sera from all three species reacted neither by ELISA nor by immunoblot with nonpathogenic treponemes, nor did guinea pig sera react with rabbit proteins. Regardless of the species of treponema used for initial adsorption, none by itself could effectively remove all cross-reactivities. Nine pathogen-specific polypeptides (15, 17, 33, 37, 39, 43, 45, 47, 97 kDa), eight (15-47 kDa) known as integral membrane or endoflagellar components, were commonly recognized by the tree species when chancre immunity developed. The substantial reduction in immune reactivity after adsorption suggests that most of these peptides consist of specific and common treponemal epitopes. Throughout the various stages of the human disease, pathogen-specific antibodies were exclusively of the IgG isotype.

Animals↗

Acute syphilitic blindness in an HIV-positive patient.

This case report describes a rare manifestation of syphilis. An HIV-positive patient with severe immunosuppression presented with rapid loss of vision due to eye involvement of Treponema pallidum infection. The ophthalmologic examination on admittance showed a chorioretinitis, an uveitis, and a swollen optical disc of both eyes. Reconstitution of vision after application of intravenous penicillin occurred within 3 weeks. Despite low HIV prevalence in central Europe, behaviors that promote transmission of HIV and other sexually transmitted diseases tend to increase. The clinical presentation of syphilis may be atypical and the rate of complications is often higher in HIV-infected patients. Thus, Treponema pallidum infection should be considered in immunosuppressed HIV-positive patients presenting with chorioretinitis. Patients should receive treatment similar to that for neurosyphilis if there is suspicion of involvement of the central nervous system.

Acute Disease↗

Morphologic analysis of enteric lesions in conventional and streptomycin-treated inbred C3H/HeN mice infected with Serpulina (Treponema) hyodysenteriae.

Oral administration of streptomycin is known to enhance the susceptibility of mice to enteric pathogens by altering the indigenous flora. We examined the effect of oral streptomycin treatment on the susceptibility of inbred C3H/HeN mice to infection with Serpulina (Treponema) hyodysenteriae. A total of 56 mice were randomly divided into four groups (A-D) of 14 each. From days 0 to 7, mice in groups A and B received streptomycin in their drinking water and mice in groups C and D served as controls. On day 7, mice in groups A and C were inoculated intragastrically with S. hyodysenteriae serotype 4, strain A1, and groups B and D served as uninoculated controls and received sterile trypticase soy broth. Clinical signs were monitored daily and body weights were recorded weekly. Mice were euthanized and necropsied for bacteriologic and histopathologic examinations on day 7 (2/group) and on days 14, 21, 28, and 35 (3/group) of the experiment. Soft fecal pellets were noticed in infected groups (A and C), but no significant differences in body weights were observed between groups (P greater than 0.05). Macroscopic changes were noted only in infected groups (A and C) beginning on day 21 of the experiment and consisted of catarrhal typhlitis, cecal emptiness, and atrophy. Histologically, the cecum and colon of mice in groups A and C had goblet cell hyperplasia, which preceded crypt epithelial cell hyperplasia, inflammatory cell infiltrates, and focal necrosis of mucosal epithelium. S. hyodysenteriae was reisolated from 10 of 12 mice in each infected group (A and C) from day 14 (7th day postinoculation) through day 35 (28th day postinoculation).(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗

Antigenic evidence for host origin of exudative fluids in lesions of Treponema pallidum-infected rabbits.

Mucoid fluid accumulating within syphilitic lesions has been considered to be of Treponema pallidum origin. To test this assumption, we examined testicular exudative fluids from T. pallidum-infected rabbits for the presence of T. pallidum antigens by various sensitive immunochemical methods, including Western blot analysis. Antigenic analysis of these fluids revealed host components but not treponemal antigens. Prolonged immunization of rabbits, guinea pigs, and a goat with this material in complete Freund adjuvant elicited low titers (fluorescent-treponemal-antibody test titer, less than or equal to 10) of antitreponemal antibodies in the rabbits and guinea pigs but not in the goat. The data suggest that these mucoid fluids are of host origin. The presence of mucopolysaccharides in these fluids may be related to the infective process. The possible mechanism by which mucopolysaccharides protect T. pallidum from immune mechanisms and its potential relationship to the pathogenesis of the disease are discussed.

Animals↗

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.

Animals↗

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↗

Effect of Treponema pallidum-infected testis supernatants on the cellular response of normal rabbit lymphocytes.

This report presents evidence indicating that both Treponema pallidum and the T. pallidum-free supernatant from T. pallidum-infected rabbit testes exert an immunosuppressive effect on the spontaneous proliferation and mitogenic response of normal rabbit lymphocytes. This effect seemed to be on the level of clonal expansion since antigenic recognition and blastogenic factor production occurred in spite of the presence of the immunosuppressive substance(s). The implication of this finding for T. pallidum multiplication in rabbit testes is discussed.

Animals↗

Ultrastructure of cells of Treponema pertenue obtained from experimentally infected hamsters.

Cells of Treponema pertenue Gauthier obtained by elution from skin lesions and lymph nodes of experimentally infected hamsters were studied in the electron microscope by means of negative staining. The cells were also examined in thin secions of skin biopsies and lymph nodes. Fimbriae were observed on the negatively stained cells. Until now fimbriae have not been demonstrated on negatively stained cells of other species of Treponema, but at present only one straon of T. pertenue has been studied in our laboratory. Otherwise, the ultrastructure of the T. pertenue cells was found to be very similar, if not identical to, the substructural details observed in cells of T. pallidum and T. cuniculi. In thin sections of skin biopsies, treponemes were observed in the intercellular spaces between cells of the stratum basale and the stratum spinosum layers of the epidermis.

Animals↗

Adoptive transfer of anti-syphilis immunity with lymphocytes from Treponema pallidum-infected guinea pigs.

Spleen and lymph node cells taken from strain 2 and strain 13 guinea pigs at the peak of their primary immune response to cutaneous syphilitic infection could transfer partial protection to symptomatic disease to normal syngeneic recipients challenged with the Nichols strain of Treponema pallidum. These recipients of immune cells had significantly fewer treponemes disseminating to the regional lymph nodes and developed fewer and less severe cutaneous lesions that resolved faster than those in guinea pigs that had been infused with normal lymphoid cells. Immune donor cells also had the capacity to transfer specific delayed-type hypersensitivity responses for T. pallidum antigens. Both T and B cells were effective in conferring anti-syphilis immunity which was associated with the almost immediate development and persistence of substantially elevated levels of circulating anti-treponemal antibody in the protected recipients. Our findings in this adoptive transfer system provide the first direct experimental evidence implicating both cellular and humoral components of the immune response as important effector mechanisms in host resistance to the pathogenic spirochete causing venereal syphilis.

Animals↗

Adoptive transfer of immunity to Treponema pallidum Nichols infection in inbred strain 2 and C4D guinea pigs.

T lymphocytes purified from lymph nodes and spleens of chancre-immune, inbred strain 2 guinea pigs, when infused into syngeneic guinea pigs, conferred protection against challenge with Treponema pallidum subsp. pallidum Nichols. No protection was conferred by similar injections of cell suspensions from normal guinea pigs or guinea pigs immunized with T. phagedenis biotype Reiter or T. pallidum-free testis supernatants from infected rabbits. Similar results were obtained with homozygous C4D guinea pigs. After several months of infection, 2 of 11 strain 2 and 1 of 8 strain C4D recipients of T. pallidum-immune cells developed an erythematous reaction of short duration at the injection site; 2 of these recipients were positive for T. pallidum. Throughout the experimental period the humoral response to treponemal antigens was substantially lower in the adoptively immune guinea pigs than in various unprotected control groups. Passive immunity to infection with T. pallidum, however, seems to be dose related, since asymptomatic infection persisted for as long as 3 months after challenge in strain 2 guinea pigs transfused with 10(8) T. pallidum-immune lymphocytes, but not in C4D recipients of twice as many immune cells.

Animals↗