Immune immobilization of Treponema pallidum: antibody and complement interactions revisited

1985 ◽  
Vol 31 (12) ◽  
pp. 1147-1151 ◽  
Author(s):  
Marianne Rice ◽  
T. J. Fitzgerald
Keyword(s):  
Divalent Cations ◽  
Treponema Pallidum ◽  
Immune Serum ◽  
Normal Serum ◽  
Rabbit Serum ◽  
Complement Cascade ◽  
Classical Complement Pathway ◽  
Heat Labile ◽  
Serum Igg ◽  
Labile Component

The Treponema pallidum immobilization test was designed for serodiagnosis of syphilis and is dependent upon specific antibody and a heat labile component of normal serum. Investigators have shown the component to be dependent upon divalent cations and it is presumed to be complement. Experiments were performed to reevaluate the interactions of antibody and complement and the mechanism of immobilization. The loss of treponemal motility was correlated to the loss of complement activity in the reaction mixture. When motility of treponemes incubated with immune serum IgG and complement had dropped to 50% (3.4 h), 72% of the available complement had been consumed. At the same time, treponemes incubated with normal serum IgG and complement were 82% motile and only 51% of the complement had been consumed. C6 deficient rabbit serum and C4 deficient guinea pig serum were used in conjunction with immune serum IgG to determine which components of the complement cascade were necessary for immobilization. Treponemes were not immobilized by either sera. Results suggest that the heat labile factor in normal sera is complement, that both early and late components of the complement cascade are necessary, and that the reaction proceeds via the classical complement pathway. Although T. pallidum is susceptible to the actions of antibody and complement, the organisms must interact with these components for at least 2 h before immobilization will result.

scholarly journals STUDIES ON TREPONEMA PALLIDUM AND SYPHILIS

1916 ◽  
Vol 23 (3) ◽  
pp. 323-328 ◽  
Author(s):  
Hans Zinsser ◽  
J. G. Hopkins
Keyword(s):  
Treponema Pallidum ◽  
Immune Serum ◽  
Normal Serum ◽  
Subsequent Publication

We believe that our experiments have shown that the serum of rabbits and sheep immunized with cultures of Treponema pallidum acquires spirochæticidal properties for these culture spirochætes. The normal serum of these animals also possesses spirochæticidal action if used in sufficient quantities, and the action of the immune serum represents probably an increase of the antibodies normally present. Both normal and immune spirochæticidal properties are destroyed by heating to 56°C. The spirochæticidal action of the immune serum can be reactivated by the addition of fresh normal serum of the same species, insufficient in amount to exert a spirochæticidal effect by itself. The structure of these spirochæticidal bodies, therefore, is entirely analogous to that of the well known bactericidal antibodies known to exist in antibacterial sera. We do not wish to have these results interpreted as applying to virulent spirochætes as well as to culture spirochætes. A subsequent publication will demonstrate why we specify this at present.

scholarly journals STUDIES ON THE SPECIFIC CHARACTERISTICS OF SYPHILITIC BLOOD PROTEINS

1931 ◽  
Vol 54 (6) ◽  
pp. 859-873 ◽  
Author(s):  
S. T. Walton
Keyword(s):  
Surface Tension ◽  
Serum Albumin ◽  
Blood Serum ◽  
Immune Serum ◽  
Normal Serum ◽  
Rabbit Serum ◽  
Blood Proteins ◽  
Buffer Solutions ◽  
Fresh Serum ◽  
Made In

These experiments show: 1. That the surface tension of normal blood serum is considerably lowered by standing undisturbed for a period of 1 hour (time-drop). 2. That the greatest time-drop recorded is with serum diluted approximately 10,000 times in fresh serum, and 50,000 times in heated serum. 3. That immune serum is not affected in the same manner by heat as is normal serum. Syphilitic serum and anti-sheep cell rabbit serum behave similarly in this respect. 4. That serum albumin is much more readily soluble in alkaline buffer solutions than globulin is, and that globulin from normal serum ionizes more than that from syphilitic serum. Further investigations are being made in an effort to determine why the proteins aggregate or dissociate under the influence of the factors under consideration.

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

1984 ◽  
Vol 160 (5) ◽  
pp. 1404-1420 ◽  
Author(s):  
S A Jones ◽  
K S Marchitto ◽  
J N Miller ◽  
M V Norgard
Keyword(s):  
Treponema Pallidum ◽  
Immune Serum ◽  
Host Cells ◽  
Rabbit Serum ◽  
Functional Activities ◽  
Molecular Weights ◽  
Rabbit Igg ◽  
Major Surface

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.

scholarly journals PROTECTIVE ANTIBODIES IN THE SERUM OF SYPHILITIC RABBITS

1939 ◽  
Vol 69 (6) ◽  
pp. 867-890 ◽  
Author(s):  
Thomas B. Turner
Keyword(s):  
Protective Action ◽  
Treponema Pallidum ◽  
Immune Serum ◽  
Normal Serum ◽  
Definite Evidence ◽  
Homologous Strain ◽  
Protective Antibodies ◽  
Partial Suppression ◽  
Series Of Experiments ◽  
Suppressive Action

1. When an emulsion containing virulent Treponema pallidum is added to serum from normal rabbits and from untreated immune syphilitic rabbits that have been infected with a homologous strain of T. pallidum the mixture incubated at 37°C., and injected intracutaneously into normal rabbits, typical syphilitic lesions commonly develop at the sites of inoculation of the normal serum-spirochete mixture, while at the sites of inoculation of immune serum-spirochete mixtures usually either no lesion develops or else the incubation period of the resulting lesions is shorter and the lesions remain smaller than those produced by normal serum-spirochete mixtures. 2. In a series of preliminary experiments, of 56 areas inoculated with serum-spirochete mixtures, in 42 the suppressive action of the syphilitic serum was manifest, in 10 areas questionable evidence of protection was noted, and in 4 areas there was no evidence that the syphilitic serum had exerted a suppressive or protective action. 3. The protective action of syphilitic serum seems to have been lessened by heating to 56°C. 4. The results of the protection test in three other series of experiments were as follows: (a) Of 12 areas in 6 rabbits inoculated with normal serum-spirochete mixtures typical syphilitic lesions developed, while in the same number of areas inoculated with immune serum-spirochete mixtures there was complete or partial suppression of lesions in all. (b) Of 45 areas inoculated with serum from 10 different immune syphilitic rabbits, definite evidence of protection was observed in 37, questionable evidence in 5, and no evidence of protection in 3. (c) Of 8 areas in 4 rabbits inoculated with immune serum-spirochete mixtures no lesions developed during the period of observation, while of 8 areas in the same rabbits inoculated with one of two normal serum-spirochete mixtures typical syphilitic lesions developed in each.

scholarly journals Immunity to the group B streptococci: interaction of serum and macrophages with types Ia, Ib, and Ic.

1976 ◽  
Vol 143 (5) ◽  
pp. 1186-1198 ◽  
Author(s):  
B F Anthony
Keyword(s):  
Group B Streptococcus ◽  
Immune Serum ◽  
Normal Serum ◽  
Rabbit Serum ◽  
Group B Streptococci ◽  
Opsonic Activity ◽  
Heat Stable ◽  
Group B ◽  
Antigen Antibody

The opsonization and phagocytosis of group B streptococci of types Ia, Ib, and Ic were studied in vitro by measuring the uptake of radioactivity by coverslip cultures of rabbit alevolar macrophages during incubation with radiolabeled, nonviable bacteria which had been exposed to rabbit serum. The uptake of counts per minute was quantitative, reproducible, and reversibly inhibited by cold, indicating that it was largely a measurement of phagocytic ingestion rather than of attachment of bacteria-immunoglobulin complexes to macrophage membranes. Moreover, suspended macrophages killed approximately 90% of viable streptococci in the presence of specific antiserum. The opsonic activity of immune serum was heat stable, and phagocytosis of streptococci was insignificant after incubation with normal serum and antiserum to some heterologous group B streptococci. By absorption studies, it was possible to identify the effect of antibodies to specific bacterial antigens. Phagocytosis of streptococci containing the corresponding antigens was maximal after opsonization with homologous or heterologous sera containing antibody to IaCHO, IbCHO, or Ibc protein. Phagocytosis of all three serotypes was intermediate when opsonization could be attributed to anti-IabcCHO. The opsonization of a specific group B streptococcus is complex and may involve two or more antigen-antibody systems.

scholarly journals THE ENHANCEMENT OF BACTERIAL PHAGOCYTOSIS BY SERUM

1969 ◽  
Vol 129 (6) ◽  
pp. 1275-1290 ◽  
Author(s):  
Richard B. Johnston ◽  
Martin R. Klemperer ◽  
Chester A. Alper ◽  
Fred S. Rosen
Keyword(s):  
Normal Serum ◽  
Phagocytic Index ◽  
Peptidase Activity ◽  
Complement Components ◽  
Serum Factors ◽  
Heat Labile ◽  
Dye Reduction ◽  
Sequential Addition ◽  
Enzymatic Inactivation

The role of serum factors in the phagocytosis of pneumococci was studied employing a spectrophotometric assay which measures reduced nitro blue tetrazolium (NBT) dye. Dye reduction occurs within the phagocyte shortly after bacterial ingestion as measured by the phagocytic index technique and by the uptake of 125I-pneumococci. Bacteria prepared with γG antibody were not phagocytosed unless a small volume of fresh normal serum was added. Using fresh sera deficient in single complement components, it was demonstrated that the first four components are necessary for optimal bacterial phagocytosis. When highly purified complement components were added to the antibody-coated pneumococci, enhancement of phagocytosis was achieved only with the sequential addition of C1, C4, C2, and C3. Evidence has been presented that human C3 bound to an immune complex exhibits peptidase activity and that this activity is essential for phagocytosis. A heat-labile, dialyzable serum cofactor which enhances C3 peptidase activity enhanced the phagocytosis of pneumococci prepared with purified complement components. A second phagocytosis-promoting cofactor, which is not a complement component, was found to be a heat-labile, 5–6S, beta pseudoglobulin. This protein may stabilize C3 peptidase activity or inhibit enzymatic inactivation of C3.

scholarly journals Regulation of complement functional efficiency by histidine-rich glycoprotein

Blood ◽  
1992 ◽  
Vol 79 (11) ◽  
pp. 2973-2980 ◽  
Author(s):  
NS Chang ◽  
RW Leu ◽  
JA Rummage ◽  
JK Anderson ◽  
JE Mole
Keyword(s):  
Divalent Cations ◽  
Alternative Pathway ◽  
Normal Serum ◽  
Peptide Sequence ◽  
Poly I:C ◽  
Complement Alternative Pathway ◽  
Nylon Membrane ◽  
Complement Components ◽  
S Protein ◽  
Functional Efficiency

The modulation of complement functional efficiency by serum histidine- rich glycoprotein (HRG) was investigated. Addition of exogenous HRG to prewarmed diluted serum, followed immediately by sensitized sheep erythrocytes (EA), resulted in enhanced hemolysis. However, when HRG was incubated with diluted serum for 10 minutes at 37 degrees C, inhibition of hemolysis occurred. The biphasic modulation of complement function was also obtained with the complement alternative pathway when HRG was added to diluted serum for hemolysis of rabbit erythrocytes. Partial reduction of complement functional activity was shown when serum was absorbed by an HRG-Sepharose 6MB column. Western blot analysis showed that complement C8, C9, factor D, and S-protein in diluted serum were bound by nylon membrane-immobilized HRG. However, by immunoprecipitation of relatively undiluted serum with anti-HRG IgG beads, HRG was found to coprecipitate with S-protein and plasminogen, which suggested that HRG may complex with these proteins in serum. In functional tests, HRG inhibited C8 hemolytic activity, probably by preventing C8 binding to EAC1–7 cells. HRG also enhanced polymerization of purified C9 as well as the generation of a 45-Kd C9 fragment. Such an effect was even more pronounced in the presence of divalent cations with the reaction mixtures of C9 and HRG. Partial dimerization of C9 was shown when exogenous HRG was added to normal serum. In contrast, polymerization of serum C9 was inhibited by exogenous HRG during poly I:C activation of serum or incubation under low ionic strength conditions. HRG was further shown to inhibit factor D-mediated cleavage of factor B when bound by cobra venom factor. The molecular basis by which HRG regulates serum complement function is not clear. Hypothetically, the tandem repetitions of a consensus histidine-rich penta-peptide sequence in HRG may provide a highly charged area that interacts with complement components.

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