scholarly journals THE EFFECT OF SALICYLATES ON THE PRECIPITATION OF ANTIGEN WITH ANTIBODY

1943 ◽  
Vol 77 (2) ◽  
pp. 173-183 ◽  
Author(s):  
Alvin F. Coburn ◽  
Eleanor M. Kapp
Keyword(s):  
Immune System ◽  
Sodium Salicylate ◽  
Sodium Tungstate ◽  
Horse Serum ◽  
Rabbit Antiserum ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Egg Albumin ◽  
Immune Precipitation ◽  
Lyotropic Series

1. Sodium salicylate modifies the precipitation of normal rabbit serum protein by sodium tungstate, and partially inhibits the precipitation of horse serum euglobulin by rabbit antiserum. Sodium salicylate added to a system containing crystalline egg albumin and its antibody partly prevents the formation of precipitate, the degree of inhibition being related to the concentration of salicylate. 2. Precipitation in the equivalence zone is more readily prevented by salicylate than precipitation in the region of antibody excess, the immune system becoming progressively less sensitive to the action of salicylate as the excess of antibody becomes larger. 3. Formed precipitates were partly dissolved following resuspension in the presence of salicylate. 4. The salicylate effect on immune precipitation is reversible, and appears to be due to inactivation of antibody. 5. Salicylate was more effective in preventing specific precipitation than other anions of a lyotropic series tested.

scholarly journals THE EFFECT OF ANTISERUM, ALONE AND WITH HYDROCORTISONE, ON FOETAL MOUSE BONES IN CULTURE

1965 ◽  
Vol 121 (4) ◽  
pp. 551-560 ◽  
Author(s):  
Honor B. Fell ◽  
L. Weiss
Keyword(s):  
Lysosomal Enzymes ◽  
Protective Action ◽  
Rabbit Antiserum ◽  
Normal Serum ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Limb Bones ◽  
Mouse Tissues ◽  
Antigen Antibody ◽  
Indirect Action

1. The effects of normal rabbit serum and of rabbit antiserum to whole foetal mouse tissues, on the isolated limb bones of late foetal mice were studied in organ culture, and the influence of hydrocortisone on these effects was investigated. 2. Unheated normal serum caused slight loss of metachromatic material from the cartilage matrix, and some resorption of both cartilage and bone. 3. In unheated antiserum to foetal mouse tissues, the terminal cartilage was smaller and less metachromatic than in paired controls in normal serum, while osteoclasis was so intense that in many explants the bone had almost disappeared. The amount of necrosis varied with different batches of antiserum. 4. The changes produced by normal serum and antiserum could be largely prevented by heating the sera to 57°C for 45 minutes. 5. The effects could also be inhibited by the addition of hydrocortisone to the unheated sera; as little as 0.1 µg hydrocortisone per ml of medium had a well marked protective action. 6. It is suggested that (a) unheated antiserum causes a release of lysosomal enzymes with consequent breakdown of intercellular material, (b) this release is due to an indirect action on the lysosome via an increased permeability of the cell membrane, (c) hydrocortisone does not affect the antigen-antibody reaction, but inhibits the autolytic changes that normally follow this reaction, possibly by stabilising both the lysosomal and cell membranes.

scholarly journals STUDIES ON PHOTO-OXIDATION OF ANTIGEN AND ANTIBODIES

1941 ◽  
Vol 73 (2) ◽  
pp. 223-242 ◽  
Author(s):  
Hans Smetana ◽  
David Shemin
Keyword(s):  
Specific Antibody ◽  
Horse Serum ◽  
Amino Nitrogen ◽  
Rabbit Serum ◽  
Type I ◽  
Egg Albumin ◽  
Photo Oxidation ◽  
Precipitin Reaction ◽  
Antigenic Properties ◽  
Chemical Studies

1. Quantitative precipitin studies indicate that progressive photo-oxidation progressively destroys the antigenic function of egg albumin. 2. Quantitative precipitin reactions of antisera (anti-egg albumin rabbit serum and antipneumococcus Type I horse serum) demonstrate that progressive photo-oxidation causes progressive lowering of the potency of the sera. 3. Quantitative precipitin reactions of the photo-oxidized globulin gamma fraction of anti-egg albumin rabbit serum and of Felton solution of antipneumococcus Type I horse serum show that these specific antibody fractions behave similarly to antibodies in whole sera. 4. Egg albumin whose precipitin reaction is destroyed by photo-oxidation no longer causes anaphylaxis in guinea pigs and does not produce precipitins in rabbits. 5. Chemical studies of progressively photo-oxidized egg albumin show a progressive destruction of tryptophane and histidine while tyrosine remains intact and cystine is reversibly oxidized. Sulfhydryl groups can no longer be demonstrated in photo-oxidized egg albumin whose antigenic characteristics are greatly weakened. 6. Similar studies on the globulin gamma fraction of anti-egg albumin rabbit serum and on Felton solution show no diminution of these amino acids in photo-oxidized material whose antigenic properties are destroyed. 7. The non-coagulable nitrogen and the amino nitrogen of egg albumin, antisera, and their specific antibody fractions show but an insignificant increase during photo-oxidation, indicating that the loss of the precipitin reaction is not due to splitting of the respective protein molecules. 8. Electrophoretic studies of egg albumin, antisera, and their specific antibody fractions show that photo-oxidation causes a marked alteration of the pattern of these substrates. 9. Photo-oxidation of proteins causes the formation of aggregates, indicating denaturation. 10. Hematoporphyrin migrates with the albumin fraction of unaltered as well as the photo-oxidized anti-egg albumin rabbit serum and pneumococcus Type I horse serum; in isolated proteins such as egg albumin, globulin gamma, or Felton solution, etc., the dye moves independently of the protein; after progressive photo-oxidation Hp becomes progressively fixed to the protein. Eosin behaves similarly to hematoporphyrin.

scholarly journals QUANTITATIVE EXPERIMENTS WITH ANTIBODIES TO A SPECIFIC PRECIPITATE. I

1941 ◽  
Vol 73 (1) ◽  
pp. 125-140 ◽  
Author(s):  
Henry P. Treffers ◽  
Michael Heidelberger
Keyword(s):  
Horse Serum ◽  
Serum Antibody ◽  
Protein Molecule ◽  
Antigenic Specificity ◽  
Rabbit Serum ◽  
Type I ◽  
Egg Albumin ◽  
Homologous Antigen ◽  
Antibody Function ◽  
Immune Rabbit

1. Rabbits were injected with the washed specific precipitate from Type II antipneumococcus horse serum. Antibody in the resulting antiserum was determined by the quantitative agglutinin method using various specific precipitates as antigens. 2. Suspensions of Types I and II antipneumococcus horse specific precipitates, as well as the specific precipitates derived from Type VIII Pn (anti-C portion), and H. influenzae horse antisera were found to remove the same amount of antibody from the immune rabbit serum. 3. Purified antibody solutions prepared by dissociation methods from Types I and II antipneumococcus horse sera were found to remove the same quantity of antibody as did the homologous specific precipitates. 4. Specific precipitates from anti-crystalline egg albumin and anti-diphtheria horse sera were found to remove only a fraction of the antibody. The reasons for this are discussed. 5. A specific precipitate prepared from pepsin-digested Type I anti-pneumococcus horse serum removed all of the antibody to the homologous antigen from the rabbit anti-precipitate serum, but followed a different quantitative course. 6. From the quantitative course of these reactions and from experiments with specific precipitates from anti-Pn rabbit and pig sera it is concluded that the only antigenic specificity demonstrable for the antibodies investigated was that due to their common origin, and that the groupings responsible for their antibody function constitute either a small part of the total protein molecule or else are non-antigenic.

scholarly journals Fluorescence cytophotometric analysis of megakaryocytic ploidy in culture: studies of normal and thrombocytopenic mice

Blood ◽  
1984 ◽  
Vol 64 (6) ◽  
pp. 1193-1199 ◽  
Author(s):  
C Chatelain ◽  
SA Burstein
Keyword(s):  
Horse Serum ◽  
Fluorescence Emission ◽  
Inverse Correlation ◽  
Cell Loss ◽  
Pokeweed Mitogen ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Cytophotometric Analysis ◽  
And Control

Abstract A system for the accurate and rapid measurement of the ploidy of cultured megakaryocytes derived from megakaryocytic colony-forming cells (CFU-M) has been developed. Thirty thousand murine marrow cells per milliliter were cultured for varying time periods in agar in the presence of horse serum and pokeweed mitogen-stimulated spleen cell- conditioned medium (PWM-SCM). To ensure the inclusion of all the megakaryocytic cells in the analysis, entire agar discs were transferred onto glass slides and dried. Cells of the megakaryocytic lineage were identified by staining for acetylcholinesterase (AchE) for two hours. Subsequently, the nuclei of the cells were stained using 1.7 X 10(-5) mol/L chromomycin A3, a specific DNA-binding fluorochrome. Megakaryocytic colonies (greater than or equal to 2 AchE+ cells) were located under transmission light. The fluorescence emission of each cell of the colony was then measured by a photometer interfaced with a computer. The mean fluorescence emission of about 20 random granulocytes per slide was used as a 2N standard. There was no significant cell loss, quenching of fluorescence by AchE staining, or overlapping of colonies or cells. Approximately 100 megakaryocytes per hour could be analyzed. Modal ploidy of cultured megakaryocytes increased from 2N to 32N between days 3 and 6 in culture. Varying concentrations of PWM-SCM from 5% to 20% did not affect the ploidy distribution when examined at day 5. The heterogeneity of the ploidy of cells within colonies increased continuously with increasing cell numbers per colony. Clonal analyses of mean ploidy and ploidy heterogeneity did not show distinct types or classes of colonies; rather, the data show that megakaryocytic colonies are structured as a continuum. An inverse correlation was found between the number of cells constituting the colonies and their mean DNA content. To determine if short-term in vivo exposure of CFU-M to a thrombocytopenic environment could affect the ploidy of their progeny, mice were given rabbit antimouse-platelet serum while control animals were given normal rabbit serum. Twenty-four hours after injection, marrow derived from these animals was cultured. At day 5, the ploidy distributions and ploidy heterogeneity were identical in both treated and control groups. Thus, factor(s) that promote CFU-M proliferation do not affect megakaryocytic endoreduplication, while stimuli that acutely influence megakaryocytic ploidy in vivo do not determine the ultimate ploidy potential of megakaryocytes derived from a CFU-M.

scholarly journals Analysis of human myelogenous leukemia cells in the fluorescence- activated cell sorter using a tumor-specific antiserum

Blood ◽  
1983 ◽  
Vol 61 (5) ◽  
pp. 858-866 ◽  
Author(s):  
AJ Malcolm ◽  
PM Logan ◽  
RC Shipman ◽  
R Kurth ◽  
JG Levy
Keyword(s):  
Bone Marrow ◽  
Acute Myelogenous Leukemia ◽  
Rabbit Antiserum ◽  
Lymphocytic Leukemia ◽  
Myelogenous Leukemia ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Cell Sorter ◽  
Chronic Granulocytic Leukemia ◽  
Acute Myelogenous

Abstract The properties of a rabbit antiserum (anti-AML) raised to a purified protein from membranes of human acute myelogenous leukemia (AML) cells is described. Bone marrow and peripheral blood leukocytes (PBL) from either normal individuals or patients with either myeloproliferative or other disorders were analyzed in a fluorescence-activated cell sorter (FACS IV) after labeling with anti-AML, normal rabbit serum (NRS), or antiserum raised to normal human membrane antigens. Of 40 cell samples from patients with acute myelogenous leukemia, 39 reacted strongly with the anti-AML antiserum. Similarly, all of 19 specimens from patients with chronic granulocytic leukemia reacted with the anti-AML. When 42 bone marrow or PBL samples from patients with a variety of lymphoproliferative disorders were examined, 2 specimens reacted with the antiserum, both from individuals with diagnoses of acute lymphocytic leukemia (ALL). None of the 14 normal bone marrow or PBL donor specimens tested reacted with the antiserum. It was also found that essentially all samples from patients in clinical remission from AML had high numbers of cells reactive with the anti-AML. When cells from such individuals were labeled and sorted on the FACS IV, it was found that cells fluorescing strongly with the anti-AML contained cells of both myeloid and lymphoid origin. The implications of these results are discussed.

scholarly journals ENHANCEMENT OF THE OPSONIZING AND AGGLUTINATING POWERS OF ANTIPNEUMOCOCCUS SERUM BY SPECIFIC PRECIPITATING SERUM

1920 ◽  
Vol 32 (3) ◽  
pp. 283-293 ◽  
Author(s):  
Ida W. Pritchett
Keyword(s):  
Horse Serum ◽  
Test Tube ◽  
Immune Serum ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Type I ◽  
Type Ii ◽  
Normal Horse Serum ◽  
Serum Type

1. No demonstrable antiopsonins are formed in rabbits following the intravenous injection of monovalent pneumococcus horse sera, Types I, II, and III. 2. The serum of rabbits injected with immune pneumococcus horse serum, Type I, II, or III, or with normal horse serum, when mixed in the proportion of 1:4 with Type I or Type II pneumococcus horse serum, can greatly augment, in vitro, the opsonization and agglutination of Type I and Type II pneumococci by the homologous immune horse sera. No similar effect is obtained with Type III serum and pneumococci. 3. The increase in opsonization and agglutination is dependent upon (a) specific sensitization of the pneumococci by the homologous immune serum and (b) the presence of the precipitating serum. In the absence of sensitization, as when a heterologous or normal horse serum is employed, opsonization and agglutination do not occur, even though a precipitating mixture is provided. The substitution of normal rabbit serum for the precipitating rabbit serum gives opsonization and agglutination in dilutions slightly higher than are effected with salt solution only, due possibly to the more favorable medium created for the leucocytes by the addition of 25 per cent of whole rabbit serum. 4. Different methods of combining the immune horse serum, precipitating rabbit serum, and pneumococci yield very similar results, preliminary sensitization of the bacteria before precipitation, or precipitation in the rabbit-horse serum mixture before the addition of the pneumococci for sensitization causing little if any difference in result from that obtained when immune horse serum, precipitating rabbit serum, and pneumococci are all mixed and incubated together. 5. This increased opsonization in the test-tube does not seem to be paralleled by increased protective power, or at any rate such protection is not readily demonstrated.

scholarly journals QUANTITATIVE EXPERIMENTS WITH ANTIBODIES TO A SPECIFIC PRECIPITATE

1942 ◽  
Vol 75 (2) ◽  
pp. 135-150 ◽  
Author(s):  
Henry P. Treffers ◽  
Dan H. Moore ◽  
Michael Heidelberger
Keyword(s):  
Horse Serum ◽  
Rabbit Antiserum ◽  
Cross Reaction ◽  
Rabbit Serum ◽  
Type Ii ◽  
Rabbit Antibody ◽  
Serum Albumins ◽  
Normal Horse Serum ◽  
Antigenic Properties ◽  
The Cross

1. Rabbit antisera to a Type II pneumococcus specific precipitate from horse serum were tested with fractions prepared by ultracentrifugation and electrophoresis of normal and immune horse serum. 2. In one instance a rapidly sedimenting protein from normal horse serum had nearly the same quantitative antigenic properties toward the anti-antibody rabbit serum as did the purified pneumococcus antibody solutions previously reported. In another instance a comparable fraction removed only a part of the rabbit antibody. 3. Electrophoretic γ-globulin from an immune horse serum had quantitatively the same antigenic properties as did antibody solutions prepared by salt-dissociation of specific precipitates. 4. Electrophoretic γ-globulin from normal horse serum differed in its antigenic behavior from γ-globulin containing antibody. The data are compared with the antigenic properties of acid and alkali treated pneumococcus specific polysaccharides toward antipneumococcus horse sera. An interpretation in terms of polymers is suggested. 5. The cross-reaction of goat serum γ-globulin against the anti-antibody serum is reported and the extent of the reaction compared with those of goat and horse serum albumins against a rabbit antiserum to the latter.

scholarly journals LOCALIZATION OF MYOGLOBIN IN HUMAN SKELETAL MUSCLE USING FLUORESCENT ANTIBODY TECHNIQUE

1967 ◽  
Vol 15 (8) ◽  
pp. 436-441 ◽  
Author(s):  
LAWRENCE J. KAGEN ◽  
RAYA GUREVICH
Keyword(s):  
Skeletal Muscle ◽  
Human Skeletal Muscle ◽  
Fluorescent Antibody ◽  
Intracellular Localization ◽  
Rabbit Antiserum ◽  
Rabbit Serum ◽  
Fluorescent Antibody Technique ◽  
Normal Rabbit Serum ◽  
Antibody Technique ◽  
Human Myoglobin

Rabbit antiserum to human myoglobin was used with the indirect fluorescent antibody technique to localize this protein in human skeletal muscle. Specific fluorescence was noted, in rapidly frozen and acetone-fixed sections, to be located at the transverse striations, at the sarcolemmal regions and at certain fibrillar structures within the cell. The antibody fluorescence reaction was shown to be specific for myoglobin, and was not produced by normal rabbit serum of serum of rabbits immunized with bacterial antigens. The reaction was abolished by prior absorption of the antimyoglobin serum with myoglobin, and was found to be absent in tissues deficient in myoglobin (lung, kidney, spleen, liver and uterus). Omission of acetone fixation or delayed freezing resulted in leakage of myoglobin from the cell and loss of specific intracellular localization. Sarcolemmal localization appeared to be somewhat more stable.

NEGATIVE FEEDBACK OF SECRETION OF FOLLICLE-STIMULATING HORMONE BY THE PITUITARY GLAND OF DEVELOPING MALE RATS

1980 ◽  
Vol 87 (3) ◽  
pp. 401-407 ◽  
Author(s):  
A. R. SHETH ◽  
GEETA R. VANAGE ◽  
A. Y. VAZE ◽  
A. N. THAKUR
Keyword(s):  
Binding Capacity ◽  
Rabbit Antiserum ◽  
Male Rats ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Human Seminal Plasma ◽  
In Vitro Binding ◽  
Vitro Binding ◽  
Age Dependent

Rabbit antiserum to human seminal plasma inhibin (hSPI) was administered subcutaneously to developing male rats of 5, 10, 14, 17 and 24 days of age and the size of the endogenous FSH rise in serum was measured. The FSH levels were threefold higher on day 9 and 1·5-fold higher on days 14 and 18 when compared with levels in control rats treated with normal rabbit serum. Furthermore, the in-vitro binding capacity of pituitary plasma membrane to 125I-labelled hSPI declined with increase in age of the rats. Thus, the results of the present study suggest that the sensitivity of the testicular inhibin–FSH feedback relationship is related to age-dependent changes in pituitary binding of inhibin.

scholarly journals INFLUENCE OF EXTRANEOUS PROTEIN AND VIRUS CONCENTRATION ON THE INACTIVATION OF THE RABBIT PAPILLOMA VIRUS BY X-RAYS

1941 ◽  
Vol 74 (5) ◽  
pp. 463-487 ◽  
Author(s):  
William F. Friedewald ◽  
Rubert S. Anderson
Keyword(s):  
Rabbit Serum ◽  
Virus Concentration ◽  
Normal Rabbit Serum ◽  
X Rays ◽  
Papilloma Virus ◽  
Egg Albumin ◽  
Buffer Solutions ◽  
Dilute Suspensions ◽  
Antiviral Antibody ◽  
Extraneous Material

The pronounced resistance to the x-rays manifested by the papilloma virus in ordinary suspensions is due to the protecting influence of extraneous matter and also in considerable degree to the amount of virus present in the preparation. Two to 4 million r were required to inactivate the virus contained in the crude papilloma extracts prepared for the present work, whereas 100,000 r or less was enough to inactivate comparable concentrations of virus after extraneous matter had been excluded by repeated differential centrifugation. The addition of normal rabbit serum or crystalline egg albumin to purified suspensions of virus was found to increase greatly the amount of irradiation required to inactivate the virus. Furthermore the percentage destruction of virus by a given amount of irradiation increases as the concentration is decreased by dilution with saline or buffer solutions. As little as 3,000 r will inactivate much of the virus in very dilute suspensions. The complement-binding antigen of papilloma virus suspensions is also inactivated by x-rays, but requires a somewhat larger amount of irradiation than necessary to destroy the infectivity of the suspensions. The effects of irradiation on the antiviral antibody present in the blood of animals which have become immune to the virus—an antibody that specifically fixes complement in mixture with the papilloma virus—are also conditioned by extraneous material. 250,000 to 500,000 r had only a slight effect on the antibody in whole serum, while this amount of irradiation completely inactivated comparable amounts of antibody in preparations partially purified by precipitation with ammonium sulfate. As a whole the findings indicate that under certain conditions of purity and concentration most of the radiation does not act by direct hits on virus or antibody particles, but indirectly by ionizing or exciting some other molecules present in the exposed suspension, which then react with the virus or antibody molecules.

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