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S Faine

Publications and source records attributed to S Faine.

At least 55 records · Page 3Linked to original sources
PubMed1984

Adhesion of leptospires to mouse fibroblasts (L929) and its enhancement by specific antibody.

The adhesion of leptospires (Leptospira interrogans serovar. copenhageni L45) to mouse L-cells was studied by microscopic observations. Within 3 h of infection of monolayers many leptospires adhered to 95-100% of the cells, and intracellular leptospires were demonstrated by electron microscopy. No specific site of attachment on the cells or the leptospires was observed. Avirulent or dead leptospires adhered poorly but attachment of the saprophytic leptospire L. biflexa serovar. patoc occurred on cell and glass surfaces. After adhesion, microvilli on the cell surfaces disappeared within 6 h of infection and cell damage was observed after 12 h. The adhesion was greatly enhanced by the presence of specific antiserum at a subagglutinating concentration. No direct penetration by leptospires of the host cells was observed with transmission and scanning electron microscopy. It appears that (1) adhesion of leptospires to L-cells precedes cell damage, and (2) leptospires may enter cells either through damaged membranes, or by a phagocytosis-like mechanism.

Adhesiveness↗
PubMed1984

Response of syphilitic rabbits to reinfection with homologous and heterologous Treponema pallidum strains.

Rabbits infected intradermally with 10(3) Treponema pallidum (Melbourne 1) cells were examined for their susceptibility to reinfection with 10(2) T. pallidum cells (homologous or heterologous strains) at various intervals after the initial infection. At 2.5 weeks after infection, the rabbits were extremely sensitive to reinfection and developed syphilitic lesions significantly faster (i.e., shorter latent periods) than control rabbits that had not received the initial infection. This phenomenon may represent a state of immunosuppression or hypersensitivity in the infected rabbits. Whatever its etiology (at present unknown), it was a transient state since at 5 weeks after infection the rabbits were no longer different from control rabbits in their susceptibility to reinfection. They showed neither immunity (i.e., longer latent periods) nor immunosuppression or hypersensitivity (shorter latent periods) upon reinfection. At 6.5 weeks after infection, two of the three experimental rabbits were fully immune (no lesions upon reinfection), whereas the other rabbit exhibited immunosuppression or hypersensitivity upon reinfection. At 7.5 and 10 weeks after infection, all of the experimental rabbits were immune to reinfection. We conclude that syphilitic rabbits show a biphasic response to reinfection, consisting of an early phase of enhanced sensitivity to T. pallidum and a later phase of immunity to T. pallidum.

Animals↗
PubMed1984

Treponema pallidum does not synthesise in vitro a capsule containing glycosaminoglycans or proteoglycans.

Treponema pallidum was investigated for its ability to synthesise glycosaminoglycans or proteoglycans in vitro. Isolated viable T pallidum organisms were incubated with radiolabelled precursors of glycosaminoglycans, sodium 35S-sulphate and 3H-glucosamine (tritiated glucosamine). T pallidum failed to incorporate sodium 35S-sulphate but did incorporate 3H-glucosamine into a macromolecule which may be associated with the surface of the treponeme. This macromolecule was resistant to degradation by specific glycosaminoglycanases. We conclude that T pallidum does not synthesise a capsule containing glycosaminoglycans in vitro.

Chromatography, Gel↗
PubMed1984

Polyanions in syphilis: evidence that glycoproteins and macromolecules resembling glycosaminoglycans are synthesised by host tissues in response to infection with Treponema pallidum.

We investigated by means of radiolabelled precursors the source and nature of the polyanionic macromolecules present in rabbit tissues during active syphilis infection. Previous studies indicated that Treponema pallidum itself does not synthesise glycosaminoglycans, at least in vitro. In replicate experiments on unilaterally infected rabbits, tissue from the orchitic testis incorporated two to three times more 35S-sulphate and 3H-glucosamine (on a wet weight basis) than tissue from the non-orchitic contralateral testis. Incorporation of 35S-sulphate was independent of the number of viable T pallidum organisms present in the infested tissue, which suggested that incorporation represented biosynthesis by the host and not the treponeme. Testes from syphilitic rabbits two days after treatment with high doses (100 mg/kg) of penicillin incorporated less 35S-sulphate than untreated infected testes, but more than normal uninfected rabbit testes. This suggests that active syphilitic infection was necessary for maximum biosynthesis of the macromolecule(s) by host tissue. Hydrodynamic profiles showed incorporation of radiolabelled precursors into two distinct fractions of different sizes, which may represent a proteoglycan and a sulphated glycoprotein. Alcian blue staining of syphilitic testes at or after peak orchitis showed focal deposition of newly synthesised polyanionic components during peak orchitis and a more generalised fibrosis in testes after peak orchitis.

Alcian Blue↗
PubMed1984

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.

Animals↗
PubMed1984

Phagocytosis of virulent and avirulent leptospires by guinea-pig and human polymorphonuclear leukocytes in vitro.

Chemiluminescence (CL) and electron microscopy were used to study the phagocytosis of both virulent and avirulent strains of Leptospira interrogans serovar copenhageni by guinea-pig and human polymorphonuclear leukocytes (PMN). A significant CL response was observed when guinea-pig PMN were incubated with virulent leptospires in the presence but not in the absence of specific immune serum. This response was markedly enhanced by the addition of guinea-pig complement. Phagocytosis was confirmed by the observation of intracellular leptospires in guinea-pig PMN by electron microscopy. The phagocytosis of avirulent leptospires by guinea-pig PMN and of both virulent and avirulent leptospires by human PMN required the presence of both specific immune serum and complement. Thus the ability of leptospires to resist phagocytosis by PMN in the absence of immune serum does not appear to be a major determinant of virulence.

Animals↗
PubMed1983

Effect of serum concentration and metabolic inhibitors on the attachment of Treponema pallidum to rabbit cells.

The effect of host-cell metabolism on the attachment of Treponema pallidum to mammalian cells in vitro was studied. The growth of baby rabbit genital organ (BRGO) cells was enhanced by increasing the concentration of serum ("serum shift-up") in the growth medium. Cells starved for 24 h in serum-free medium showed a burst of DNA synthesis when shifted to fresh medium containing 20% serum. In aerobic conditions, they were much more heavily coated with attached T. pallidum than cells shifted to 20% serum after maintenance at serum concentrations of 2.5%, 5% or 10%. This effect was very pronounced during the first few hours of co-incubation. In microaerophilic conditions, the extent of T. pallidum adherence also paralleled the increase in DNA synthesis by BRGO cells. Cycloheximide and methotrexate greatly inhibited DNA and protein synthesis in BRGO cells, but did not affect the motility of T. pallidum. When BRGO cell metabolism was inhibited by these two drugs, attachment of T. pallidum was significantly decreased. These results indicate that T. pallidum attaches best to actively growing BRGO cells in tissue culture. This may explain the apparently preferential parasitism of actively growing tissues by T. pallidum in syphilis in man.

Adhesiveness↗
PubMed1983

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.

Animals↗
PubMed1983

A pomona serogroup-specific, agglutinating antigen in Leptospira, identified by monoclonal antibodies.

Monoclonal antibodies were produced by hybridoma cell lines derived by fusion of mouse NS-1 myeloma cells with splenocytes from mice immunized with Leptospira interrogans serovar pomona. One hybridoma (A3) produced an IgG2a antibody which agglutinated all leptospires of the Pomona serogroup but not leptospires representative of serovars of any other serogroup. The antibody precipitated in immunodiffusions with either alkali- or phenol-extracted lipopolysaccharide and also with the TM antigen of Yanagawa et al., indicating a common determinant on both antigens. A3 antibody opsonized viable leptospires for phagocytosis by mouse macrophages in vitro.

Animals↗
PubMed1983

A monoclonal IgM smooth muscle antibody reactive with fibroblast stress fibres produced by immunization with Treponema pallidum.

A monoclonal IgM smooth muscle antibody secreted by a hybrid (MMI-1) of mouse plasmacytoma NS-1 with spleen cells from mouse immunized with Treponema pallidum was detected by indirect immunofluorescence tests on frozen tissue sections and on acetone fixed monolayers of rat and human fibroblasts. The antibody did not react with acetone fixed smears of T. pallidum but reacted with smooth muscle fibres and with striations of skeletal and cardiac muscle. In non-muscle cells, the antibody stained liver in a 'polygonal' pattern, thymus with accentuated staining of the thymic medulla, renal glomeruli and the brush border and peritubular fibrils of renal tubules. In fibroblast monolayers, the antibody stained stress fibres in an interrupted pattern. Immunoblotting with muscle proteins and the antibody showed labelling of a 100K molecule. The cellular distribution of the mouse monoclonal antibody is similar to that obtained with anti-actin antibody suggesting that the corresponding antigen may be an actin binding protein.

Actins↗
PubMed1983

Species- and genus-specific antigens in Leptospira, revealed by monoclonal antibodies and enzyme immunoassay.

Mouse antisera to Leptospira interrogans serovar pomona or L. biflexa serovar patoc agglutinated homologous leptospires but not leptospires of serovars from any heterologous serogroup tested. When tested by enzyme immunoassay (EIA), pomona and patoc antisera reacted with all serovars tested from L. interrogans, L. biflexa or L. illini species. A non-agglutinating hybridoma-derived monoclonal antibody reacted by EIA only with serovars of the pathogenic L. interrogans species but not with L. biflexa nor L. illini. The results demonstrated the presence of both genus and species specific non-agglutinating leptospiral antigens which could be detected only by EIA.

Antibodies, Monoclonal↗
PubMed1982

Bovine IgM and IgG response to Leptospira interrogans serovar hardjo as measured by enzyme immunoassay.

The enzyme-linked immunosorbent assay (ELISA) was used to detect specific IgG and IgG antibodies in the sera of cattle infected or immunized with Leptospira interrogans serovar hardjo. IgM appeared first but was quickly followed by IgG which persisted longer than IgM. The levels of antibody detectable by ELISA and by the microscopic agglutination test (MAT) did not correlate, suggesting that the two techniques measured different antigen--antibody systems. The transient nature of the IgM response as measured by ELISA indicates potential usefulness as a serodiagnostic test for detecting current leptospiral infections.

Agglutination Tests↗
PubMed1982

Evidence that leptospiral lipopolysaccharide is not an important protective antigen.

CBA/N mice, which could not produce antibodies against lipopolysaccharide (LPS) from either Escherichia coli or Leptospira interrogans serovar pomona, produced levels of agglutinating antibodies against leptospires similar to those produced by immunologically normal CBA mice. CBA/N mice were thus resistant to acute leptospiral infection and CBA/N immune serum passively protected immunosuppressed mice from infection. The results suggest that antibodies against LPS are not important in protection against experimental leptospiral infection in mice.

Animals↗
PubMed1982

Immunogenicity of leptospiral vaccines grown in protein-free medium.

Leptospira interrogans serovars pomona and hardjo were adapted to grow in a chemically-defined, protein-free (PF) medium. Formolised monovalent vaccines of serovars pomona and hardjo and a bivalent mixture of the two were prepared from PF cultures. Live PF cultures and the vaccine preparations retained their agglutinating antigens and their immunogenicity when tested in rabbits and guinea-pigs. The vaccines were not pyrogenic and dermal reactions were slight.

Agglutinins↗
PubMed1982

The role of macrophages in the protection of mice against leptospirosis: in vitro and in vivo studies.

Balb/c mice are naturally resistant to infection with Leptospira interrogans serovar copenhageni, but leptospires were not phagocytosed by mouse peritoneal macrophages in vitro without added specific antibody. Similar results were obtained irrespective of whether leptospires were viable or killed, virulent or avirulent, or whether macrophages were obtained from normal mice, immunized mice or mice previously infected with BCG. Suppression or stimulation of macrophage function in vivo did not affect the outcome of infection of immunosuppressed mice with leptospires; specific antibody was essential for protection from infection.

Animals↗
PubMed1982

The sensitivities of different immunoassays for detecting leptospiral antigen.

The detection of leptospiral antigen in biological fluids is important for the diagnosis of leptospirosis in animals and man. However the sensitivity of dark field microscopy, the usual detection method, is often inadequate. A comparison was made between the sensitivities of several immunological techniques for detecting Leptospira interrogans serovar hardjo. By staphylococcal coagglutination 10(8) leptospires per ml could be detected and by countercurrent immunoelectrophoresis 10(7) per ml. The best sensitivity obtained by enzyme-linked immunosorbent assay was 10(5) leptospires per ml, and by radioimmunoassay 10(4) to 10(5) per ml. Radioimmunoassay offers the prospect of improved diagnosis of leptospirosis through the detection of leptospiral antigen.

Agglutination Tests↗
PubMed1981

The enzyme-linked immunosorbent assay (ELISA) as a serological test for detecting antibodies against Leptospira interrogans serovar hardjo in sheep.

The enzyme-liked immunosorbent assay (ELISA) was compared with the standard microscopic agglutination test (MAT) as a method for detecting antibodies against Leptospira interrogans serovar hardjo in sheep. Peak antibody levels detected by the 2 tests occurred at different times following experimental infection of sheep. In serums from flocks of sheep with naturally acquired infection there was a 95% correlation between MAT and ELISA with respect to the presence or absence of antibody to serovar hardjo, although the levels of correlation of the titres of the 2 tests was low. The 2 tests appeared to measure different antigen-antibody systems. The ELISA would be a useful test for screening large numbers of serums for antibodies to L. interrogans serovar hardjo.

Agglutination Tests↗