STUDIES ON A THYROID STIMULATING FACTOR IN URINARY CHORIONIC GONADOTROPHIN PREPARATIONS

1967 ◽  
Vol 55 (4) ◽  
pp. 587-599 ◽  
Author(s):  
A. Burger ◽  
R. Kunz
Keyword(s):  
Biological Response ◽  
Chorionic Gonadotrophin ◽  
Rabbit Serum ◽  
Soluble Antigen ◽  
Globulin Fraction ◽  
Human Pituitary ◽  
Antibody Complex ◽  
Antigen Antibody ◽  
Stimulating Factor ◽  
Immune Rabbit

ABSTRACT Thyroid stimulating activity was demonstrated in human urinary chorionic gonadotrophin preparations (HCG) by the method of McKenzie (1958). The biological response of the material was similar to that of bovine pituitary thyrotrophin (TSH), but no neutralization of the activity occurred with antibodies against human pituitary TSH and, even more interesting, with antibodies against human urinary chorionic gonadotrophin preparations. The latter antibodies perfectly neutralized human pituitary TSH. The Gamma-G-globulins precipitated from a mixture of HCG and anti-HCG contained biological activity whereas no activity was recovered in the Gamma-G-globulin fraction when HCG was mixed with non immune rabbit serum. This may indicate the presence of a soluble antigen-antibody complex.

scholarly journals THE PROTECTIVE ACTION OF TYPE I ANTIPNEUMOCOCCUS SERUM IN MICE

1936 ◽  
Vol 64 (3) ◽  
pp. 377-383 ◽  
Author(s):  
Kenneth Goodner ◽  
Frank L. Horsfall
Keyword(s):  
Selective Adsorption ◽  
Protective Action ◽  
Horse Serum ◽  
Rabbit Serum ◽  
Type I ◽  
Antibody Complex ◽  
Antigen Antibody

1. The addition of small amounts of cholesterol and of cephalin reduces markedly the protective action of antipneumococcus horse serum. 2. These lipids do not affect the protective action of antipneumococcus rabbit serum. 3. These findings may be explained (a)by the selective adsorption of lipid on the antigen-antibody complex, and (b) by certain lipid antagonisms. 4. The failure of large amounts of immune horse serum to protect mice against pneumococcus infection is explicable on the basis of selective participation of lipids dependent upon the species from which the antibody is derived. 5. The lipids modify the results of protection tests only through participation in the process of specific sensitization.

scholarly journals THE UNLABELED ANTIBODY ENZYME METHOD OF IMMUNOHISTOCHEMISTRY PREPARATION AND PROPERTIES OF SOLUBLE ANTIGEN-ANTIBODY COMPLEX (HORSERADISH PEROXIDASE-ANTIHORSERADISH PEROXIDASE) AND ITS USE IN IDENTIFICATION OF SPIROCHETES

1970 ◽  
Vol 18 (5) ◽  
pp. 315-333 ◽  
Author(s):  
LUDWIG A. STERNBERGER ◽  
PAUL H. HARDY ◽  
JOHN J. CUCULIS ◽  
HOWARD G. MEYER
Keyword(s):  
Horseradish Peroxidase ◽  
Rabbit Antiserum ◽  
Antibody Fragment ◽  
Soluble Antigen ◽  
Simple Method ◽  
Antibody Complex ◽  
High Yields ◽  
Antigen Antibody ◽  
Unlabeled Antibody ◽  
Specific Rabbit

Antigen was identified histochemically without the use of labeled antibodies by the sequential application of (a) specific rabbit antiserum, (b) sheep antiserum to rabbit immunoglobulin G, (c) specifically purified, soluble horseradish peroxidase-anti-horseradish peroxidase complex (PAP), (d) 3,3'-diaminobenzidine and hydrogen peroxide and (e) osmium tetroxide. A simple method for preparation of high yields of PAP consisted of precipitation of antibody from specific rabbit antiserum with horseradish peroxidase (PO) at equivalence, solubilization of the washed precipitate with excess PO at pH 2.3, 1°C, followed by immediate neutralization and separation of PAP from PO by half-saturation with ammonium sulfate. The ratio of PO to anti-PO in PAP was 3:2 irrespective of the source of antiserum. PAP was heterogeneous on electrophoresis, homogeneous on sedimentation, diffusion and electron microscopy and consisted of pentagons with diameters of 205 Å. s20, w, 11.98 x 10–13; d20, w, 2.48 x 10–7; molecular weight by sedimentation velocity, 429,000, and equilibrium, 413,000. Sensitivity and specificity of immunohistochemical staining of spirochetes was about 100- to 1000-fold that of immunofluorescence. The unexpected ratio of PO to anti-PO is presumed to be due to stabilization by the pentagonal shape in which three corners are suspected to be PO and two antibody fragment Fc.

scholarly journals STUDIES IN THE SEROLOGY OF SYPHILIS

1930 ◽  
Vol 52 (5) ◽  
pp. 739-746 ◽  
Author(s):  
Harry Eagle
Keyword(s):  
Complement Fixation ◽  
Essential Feature ◽  
Forthcoming Paper ◽  
Red Cells ◽  
Globulin Fraction ◽  
Serum Globulin ◽  
Flocculation Test ◽  
Antibody Complex ◽  
Wassermann Reaction ◽  
Antigen Antibody

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.

scholarly journals THE PNEUMOCOCCAL CAPSULAR SWELLING REACTION, STUDIED WITH THE AID OF THE ELECTRON MICROSCOPE

1943 ◽  
Vol 78 (5) ◽  
pp. 327-332 ◽  
Author(s):  
Stuart Mudd ◽  
Ferdinand Heinmets ◽  
Thomas F. Anderson
Keyword(s):  
Capsular Polysaccharide ◽  
Specific Antigen ◽  
Immune Serum ◽  
Rabbit Serum ◽  
Surface Deposit ◽  
Specific Serum ◽  
Antibody Complex ◽  
Wall Interaction ◽  
Antigen Antibody ◽  
Serum Components

Electron micrographs indicate, in harmony with previous findings, that the pneumococcal capsule is a gel of low density outside of and closely applied to the bacterial cell wall. Interaction with homologous immune rabbit serum greatly increases the thickness and density of this capsular gel; the increase in thickness of the specifically swollen pneumococcal capsule may exceed by 25-fold the thickness of the surface deposit caused by rabbit immune serum on the cell walls and flagella of homologous non-capsulated bacteria. Conclusions drawn from these and earlier data are that homologous immune serum permeates the pneumococcal capsular gel; the specific antibody combines with the capsular polysaccharide; non-specific serum components are secondarily adsorbed to or combined with the specific antigen-antibody complex. The relatively low antibacterial titers characteristic of pneumococcal antisera can be explained in part by the permeation of the capsule by antiserum, in part by the high combining capacity of pneumococcal carbohydrate for antibody (17).

The absorption of antibody from immune sera and from mixtures of sera by the gut of the young rat

1958 ◽  
Vol 148 (930) ◽  
pp. 92-103 ◽  
Keyword(s):  
Marked Effect ◽  
Immune Serum ◽  
Maximum Amount ◽  
Rabbit Serum ◽  
Antibody Concentration ◽  
Globulin Fraction ◽  
Rabbit Antibody ◽  
Rat Serum ◽  
Old Rats ◽  
Immune Rabbit

The amount of antibody transmitted to the circulation from immune rat serum administered by mouth in a single dose to 12-day-old rats is directly proportional both to the antibody concentration and to the volume of serum administered for doses up to 0⋅10 ml. The amount of antibody transmitted does not increase with larger doses and is limited to that sufficient to produce a mean circulating titre of 1/32 of that of the immune serum. The uptake of the antibody-containing globulin fraction from a single feed is sufficient to account for the whole of the increase in this fraction of the young rats’ serum protein over a period of at least 9 h. The proportion of antibody transmitted to the circulation from immune rabbit serum administered by mouth declines nearly linearly with increasing volume of dose up to the maximum that could be administered of 0⋅60 ml. The proportion of rabbit antibody transmitted is approximately 1/8 of that of rat antibody from corresponding doses of immune serum up to 0⋅10 ml. Admixture of certain heterologous sera with the immune rat or rabbit serum administered by mouth interferes with the transmission of antibody to the circulation to an extent that cannot be accounted for by the dilution. Human, ox, guinea-pig and rabbit sera have a marked effect, whereas sheep, mouse, hamster and rat sera have little or no effect. Interference with the transmission of rat antibodies results even when the volume of the dose of mixed sera is less than 0⋅10 ml. Although interference decreases the proportion of antibody transmitted to the circulation it does not reduce significantly the maximum amount that can be transmitted.

FURTHER OBSERVATIONS ON THE IMMUNOLOGICAL BEHAVIOUR OF HUMAN PITUITARY AND CHORIONIC GONADOTROPHINS

1964 ◽  
Vol 29 (3) ◽  
pp. 263-271 ◽  
Author(s):  
G. ILLEI ◽  
PAMELA M. MORITZ
Keyword(s):  
Human Serum ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Normal Human Serum ◽  
Globulin Fraction ◽  
Human Pituitary ◽  
Human Menopausal Gonadotrophin ◽  
Normal Human ◽  
Antigenic Factor

SUMMARY Antisera were raised to human chorionic gonadotrophin (HCG) and human menopausal gonadotrophin (HMG). Normal human serum and human pituitary and chorionic gonadotrophins reacted with both antisera in immunological experiments. After absorption with human serum, the γ-globulin fractions of the antisera were isolated, and only the reaction to the hormone preparations remained. Immunoelectrophoresis revealed a common antigenic factor in the α2-β globulin region. The γ-globulin fraction of HCG antiserum seems to be suitable for the assay of HCG, while that of the HMG antiserum might be employed for the detection of HMG.

scholarly journals The conglutination phenomenon. IV. An experimental investigation of the factors determining the adsorption of complement by an antigne-antiserum mixture

1950 ◽  
Vol 48 (1) ◽  
pp. 73-95 ◽  
Author(s):  
A. M. Blomfield ◽  
R. R. A. Coombs ◽  
N. H. Hole
Keyword(s):  
General Pattern ◽  
Complex Problem ◽  
Specific Factor ◽  
Soluble Antigen ◽  
Particulate Antigen ◽  
Two Factors ◽  
Initial Intervention ◽  
Antibody Complex ◽  
The One ◽  
Antigen Antibody

1. Salmonella antisera from a number of animals, including man, have been examined by conglutinating complement-absorption and haemolytic complement-fixation tests for the presence of anitbodies. Both a soluble antigen and a particulate antigen of washed suspended orgainsms have been used. The behavior of the antisera was comparable with that previously recorded with the mallein antimallein system, in so far as the reactions obtained with the different complements followed the same general pattern.2. Two factors were found to explain the failure of certain antisera, in conjunction with the antigen, to adsorb certain complements. On the one hand, a few antibodies themselves appeared incapable of fixing a particular complement. On the other hand, although some antisera failed to fix specific complements, their antibodies, isolated from the serum, were able to fix the complements concerned. In these cases it appeared that the heated serum contained a non-specific factor that prevented the adsorption of the complements.3. The existence of such non-specific factors in certain heat-inactivated sera is illustrated in Figs. 11–14. An examination of the mechanism involved does not appear to implicate the procomplementary effect of sera, although this effect in itself is a complex problem requiring further elucidation. The few experiments carried out indicate that the factor in the inactivated serum does not come into play and inhibit the adsorption of complement by an antigen-antibody complex without the initial intervention of some component of complement. Only after this has occurred is the inhibitory factor able to block any further complement adsorption.4. We have discussed the implications of these experiments and their possible relationships to the complementoid phenomena described by previous workers.

Visualization of Antibody Transport in Rat Visceral Yolk-Sac Placenta

1982 ◽  
Vol 40 ◽  
pp. 246-247
Author(s):  
William P. Jollie
Keyword(s):  
Phosphate Buffer ◽  
Yolk Sac ◽  
Female Rats ◽  
Thin Sections ◽  
Visceral Yolk Sac ◽  
Antibody Complex ◽  
Cytochemical Reaction ◽  
Hind Foot ◽  
Antigen Antibody ◽  
Yolk Sac Placenta

A technique has been developed for visualizing antibody against horseradish peroxidase (HRP) in rat visceral yolk sac, the placental membrane across which passive immunity previously has been shown to be transferred from mother to young just prior to birth. Female rats were immunized by injecting both hind foot pads with 1 mg HRP emulsified in complete Freund's adjuvant. They were given a booster of 0.5mg HRP in 0.1 ml normal saline i.v. after one week, then bred and autopsied at selected stages of pregnancy, viz., 12, 1 7 and 22 days post coitum, receiving a second booster, injected as above, five days before autopsy. Yolk sacs were removed surgically and fixed immediately in 2% paraformaldehye, 1% glutaraldehye in 0.1 M phosphate buffer with 0.01% CaCl2 at pH 7.4, room temperature, for 3 hr, rinsed 3X in 0.1 M phosphate buffer plus 5% sucrose, then exposed to 1 mg HRP in 1 ml 0.1 M phosphate buffer at pH 7.4 for 1 hr. They were refixed in aldehydes, as above, for 1 5 min (to assure binding of antigen-antibody complex). Following buffer washes, the tissues were incubated in 3 mg diaminobenzidine tetrahydrochloride and 0.01% H2O2 in 0.05 M Tris-HCl buffer for 30 min. After brief buffer washes, they were postfixed in 2% OsO4. in phosphate buffer at pH 7.4, 4°C for 2 hr, dehydrated through a graded series of ethanols, and embedded in Durcupan. Thin sections were observed and photographed without contrast-enhancement with heavy metals. Cytochemical reaction product marked the site of HRP (i.e., antigen) which, in turn, was present only where it was bound with anti-HRP antibody.

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