scholarly journals Double immunocytochemical staining in the study of antibody-producing cells in vivo. Simultaneous detection of cells producing monospecific antibodies and cells producing cross-reacting antibodies in the spleen of rabbits after injection of two related antigens.

1984 ◽  
Vol 32 (8) ◽  
pp. 844-848 ◽  
Author(s):  
N Van Rooijen ◽  
N Kors ◽  
R Van Nieuwmegen
Keyword(s):  
Gamma Globulin ◽  
Simultaneous Detection ◽  
Immunocytochemical Staining ◽  
Secondary Immune Response ◽  
Spleen Tissue ◽  
Specific Antibodies ◽  
Similarities And Differences ◽  
Human Gamma Globulin ◽  
Monospecific Antibodies

Rabbits were injected simultaneously with both human gamma globulin (HGG) and bovine gamma globulin (BGG). Sections of spleen tissue were prepared from spleen biopsies taken during the primary or secondary immune response, and incubated simultaneously with horseradish peroxidase (HRP)-HGG conjugate and alkaline phosphatase (AP)-BGG conjugate in order to detect cells containing specific antibodies against one or both of the antigens. After both HRP and AP cytochemistry, cells with a red-stained cytoplasm, cells with a blue-stained cytoplasm, and cells with a violet-stained cytoplasm were detected in the spleen. The red-stained cells had bound the HRP-HGG conjugate, indicating that these cells contained anti-HGG antibodies. The blue-stained cells had bound the AP-BGG conjugate, indicating that these cells contained anti-BGG antibodies. The violet-stained cells had obviously bound both the HRP-HGG conjugate and the AP-BGG conjugate, indicating that these cells contained antibodies cross-reacting with both antigens. Results are compared with earlier studies on the antigenic similarities and differences between HGG and BGG when used as antigens in rabbits.

scholarly journals Double immunocytochemical staining in the study of antibody-producing cells in vivo. Combined detection of antigen specificity (anti-TNP) and (sub)class of intracellular antibodies.

1985 ◽  
Vol 33 (3) ◽  
pp. 175-178 ◽  
Author(s):  
N Van Rooijen ◽  
N Kors
Keyword(s):  
Keyhole Limpet Hemocyanin ◽  
Staining Technique ◽  
Immunocytochemical Staining ◽  
Antigen Specificity ◽  
Specific Antibodies ◽  
Igm Antibodies ◽  
Rabbit Class ◽  
Spleen And Lymph Nodes ◽  
Combined Detection

Mice and rabbits were immunized with trinitrophenyl (TNP)-conjugated keyhole limpet hemocyanin (KLH). Cells producing specific antibodies against the hapten TNP were detected in vivo in spleen and lymph nodes using a TNP--alkaline phosphatase (AP) conjugate. Using horseradish peroxidase (HRP)-conjugated anti-mouse (sub)class (IgG2A, IgG2B, IgM) antibodies and anti-rabbit class (IgG, IgM) antibodies and a double immunocytochemical staining technique for simultaneous demonstration of the enzymes AP and HRP, we were able to determine both the antigen specificity (anti-TNP) and the (sub)class of intracellular antibodies produced by individual antibody-forming cells in vivo.

scholarly journals REGULATION OF ANTIBODY FORMATION BY SERUM ANTIBODY

1969 ◽  
Vol 130 (5) ◽  
pp. 1175-1186 ◽  
Author(s):  
Martin W. Graf ◽  
Jonathan W. Uhr
Keyword(s):  
Antibody Formation ◽  
Gamma Globulin ◽  
Regulatory Mechanism ◽  
Serum Antibody ◽  
Gel Filtration ◽  
Antibody Activity ◽  
Specific Antibodies ◽  
Human Gamma Globulin ◽  
Antibody Levels ◽  
Conventional Antibody

Rabbits were injected intravenously with bovine serum albumin (BSA) and bacteriophage T2 (T2). 2–3 wk later, anti-BSA was removed from such animals by a procedure which involved exposure of removed plasma to an immunoadsorbent (125I-BSA bound to bromoacetyl cellulose) and return of the adsorbed plasma to the animal. This resulted in removal of the majority of antibody activity to BSA without affecting antibody levels to T2. 1–2 days later, anti-BSA levels began to rise, and reached peak levels usually 5 days after the removal of antibody. Antibody levels to T2 did not change. No evidence was obtained that BSA was released from the immunoadsorbent into the circulation of the rabbits. Thus, only trace amounts of radioactivity were released into the plasma; most of the radioactivity was equally coprecipitable with BSA or human gamma globulin and their specific antibodies; the released material was not demonstrated to be immunogenic in primed rabbits; and the released material did not elute with BSA on gel filtration. The results are interpreted as evidence that serum antibody acts as a regulatory mechanism for antibody formation during the conventional antibody response to a metabolizable antigen.

THE METABOLISM OF ANTIGEN IN VIVO: II. AZOPROTEIN

1964 ◽  
Vol 42 (6) ◽  
pp. 833-839
Author(s):  
A. Larose ◽  
B. Rose ◽  
M. Richter
Keyword(s):  
Intravenous Injection ◽  
Experimental Animal ◽  
Gamma Globulin ◽  
Double Diffusion ◽  
Single Intravenous Injection ◽  
Serum Fractions ◽  
Human Gamma Globulin

Experiments were carried out to detect precipitating or non-precipitating antigenic fragments in the serum or spleen of rabbits given a single intravenous injection of arsanil azo – human gamma-globulin. The results, using a number of different methods for the detection of antibody (double diffusion, immunoelectrophoresis, inhibition of hemagglutination by serum fractions obtained by elution of paper electropherograms of the antiserum), failed to provide any evidence for the presence of antigenic fragments either in the circulation or in the spleen of the experimental animal.

Effect of ragweed hyperimmune human gamma globulin on in vivo and in vitro parameters of ragweed sensitivity

1976 ◽  
Vol 57 (4) ◽  
pp. 335-341 ◽  
Author(s):  
Elliot Rubinstein ◽  
Yosuke Fujita ◽  
Tadao Okazaki ◽  
Daniel Tripodi ◽  
Robert E. Reisman ◽  
...  
Keyword(s):  
Gamma Globulin ◽  
Human Gamma Globulin

scholarly journals Chronic Lymphocytic Leukemia, Hypogammaglobulinemia and Autoimmune Hemolytic Anemia— An Experiment of Nature

Blood ◽  
1960 ◽  
Vol 15 (5) ◽  
pp. 748-757 ◽  
Author(s):  
ANTHONY V. PISCIOTTA ◽  
LOUIS F. JERMAIN ◽  
JEAN E. HINZ
Keyword(s):  
Hemolytic Anemia ◽  
Autoimmune Hemolytic Anemia ◽  
Gamma Globulin ◽  
Lymphocytic Leukemia ◽  
Chronic Lymphatic Leukemia ◽  
Lymphatic Leukemia ◽  
Normal Human ◽  
Human Gamma Globulin

Abstract 1. The case study is presented of a 75 year old man who had chronic lymphatic leukemia, autoimmune hemolytic anemia and hypogammaglobulinemia. The positive antiglobulin reaction with serum made from gamma globulin and the neutralization of the antiglobulin reaction with human gamma globulin demonstrated that this patient’s erythrocytes were coated with gamma globulin. 2. There was a normal survival time of I131-labeled normal human gamma globulin, suggesting defective synthesis of gamma globulin. Failure to demonstrate radioactivity on the patient’s erythrocytes when I131-labeled normal gamma globulin was given signified that normal human gamma globulin has no affinity in vivo for the patient’s red cells and that the erythrocyte-coating protein was derived from a source endogenous to the patient. 3. These relationships favor an immunologic mechanism in the development of an antiglobulin reaction in this patient.

scholarly journals THE USE OF PRECIPITIN ANALYSIS IN AGAR FOR THE STUDY OF HUMAN STREPTOCOCCAL INFECTIONS

1958 ◽  
Vol 108 (3) ◽  
pp. 385-410 ◽  
Author(s):  
Seymour P. Halbert
Keyword(s):  
Gamma Globulin ◽  
Group A Streptococcus ◽  
Streptococcal Infections ◽  
High Potency ◽  
Streptolysin O ◽  
Combined Use ◽  
Group A ◽  
Human Gamma Globulin ◽  
Human Infections

As evidenced by precipitin analysis with pooled human gamma globulin, at least 12 distinct antigens were produced in cultures by one strain of Group A streptococcus (C203S). It was suggested on this basis, that these antigens were produced in vivo during human infections. By the combined use of continuous flow electrophoresis on paper curtains, and column chromatography with calcium phosphate gels, five of these have been isolated in a probable high state of purity. One of the components was obtained from culture filtrates of a Group C streptococcal strain. Three of the purified antigens have been tentatively identified as streptolysin "O", diphosphopyridinenucleotidase, and proteinase precursor. The latter could be very readily crystallized, and appears "identical" with that described by Elliott. The DPNase was of extremely high potency, 1 mg. being capable of destroying 12.6 gm. of DPN in 7½ minutes at 37°C. The identity of the other two components is uncertain as yet. They are distinct from each other and the above products immunologically, and are not related to the "C" carbohydrate. The applicability of these methods for the analysis of infectious diseases generally was discussed.

scholarly journals Double immunocytochemical staining in the study of antibody-producing cells in vivo. Detection of specific antibody-producing cells in the spleen and simultaneous determination whether or not they produce immunoglobulin G antibodies.

1984 ◽  
Vol 32 (7) ◽  
pp. 677-680 ◽  
Author(s):  
N Van Rooijen ◽  
N Kors ◽  
R Van Nieuwmegen
Keyword(s):  
Alkaline Phosphatase ◽  
Immunocytochemical Staining ◽  
Igg Antibodies ◽  
Specific Antibodies ◽  
In Vivo Detection ◽  
Rabbit Igg ◽  
Present Technique ◽  
Antibody Class ◽  
Ig G

Rabbits were primed intravenously with human serum albumin (HSA) and boosted with the same antigen 2 months later. Cells producing specific antibodies against HSA could be detected in vivo and it could be determined whether or not they belonged to the immunoglobulin (Ig) G class using a combined peroxidase (HRP) and alkaline phosphatase (AP) immunocytochemical technique. HRP-HSA conjugate was used for detection of anti-HSA-producing cells and AP-sheep anti-rabbit IgG (SRIgG) was used to determine the IgG class of the antibodies produced by these cells in the same spleen section. After performing both HRP and AP cytochemistry, cells with a red-stained cytoplasm represent anti-HSA-producing cells not stained for their antibody class and cells with a blue-stained cytoplasm represent cells producing IgG antibodies not directed against HSA. Cells with a double-stained cytoplasm represent cells producing anti-HSA antibodies belonging to the IgG class. We also attempted to determine whether or not part of the anti-HSA-producing cells belonged to the IgM class using AP-sheep anti-rabbit IgM (SRIgM). In this case no double-stained cells were detected, indicating that the affinity of intracellular IgM-anti-HSA antibodies is too low to allow detection using the present technique.

scholarly journals Regulation of the immune response. I. The potentiation of in vivo and in vitro immune responses by Fc fragments

1980 ◽  
Vol 152 (1) ◽  
pp. 113-123 ◽  
Author(s):  
EL Morgan ◽  
SM Walker ◽  
ML Thoman ◽  
WO Weigle
Keyword(s):  
Immune Response ◽  
Immune Responses ◽  
Gamma Globulin ◽  
Adjuvant Effect ◽  
Sheep Erythrocytes ◽  
Human Gamma Globulin

Fc fragments derived from human and murine Ig were found to be potent adjuvants when administered with antigen. Both the in vivo and in vitro anti- sheep erythrocytes (SRBC) responses were significantly enhanced by Fc fragments. The adjuvant effect was shown to be extremely dependent upon the dose of antigen used, with the greatest enhancement occurring when suboptimal doses of antigen are employed. The anti-genicity of the Fc molecule was not related to its adjuvanticity because homologous Fc was as potent an adjuvant as heterologous Fc. Moreover, human Fc fragments enhanced anti-SRBC responses in mice which were tolerant to human gamma globulin.

THE METABOLISM OF ANTIGENS IN VIVO: I. PROTEIN ANTIGENS

1964 ◽  
Vol 42 (6) ◽  
pp. 821-832
Author(s):  
M. Richter ◽  
C. Larose ◽  
B. Rose
Keyword(s):  
Human Serum Albumin ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Gamma Globulin ◽  
Double Diffusion ◽  
Protein Antigens ◽  
Single Intravenous Injection ◽  
Human Gamma Globulin ◽  
Antigen Interaction

Attempts were made to detect antigenic fragments in the sera or spleens of rabbits given a single intravenous injection of bovine serum albumin, human gamma-globulin, or human serum albumin. The results obtained, using a number of different methods for the detection of antibody–antigen interaction (double diffusion in gel, immunoelectrophoresis, inhibition of hemagglutination), failed to provide any evidence in favor of the presence of antigenic fragments either in the circulation or in the spleen of the experimental animal.

Protective effects of S-sulfonated human gamma globulin against experimental infections in mice

1980 ◽  
Vol 30 (2) ◽  
pp. 329-336
Author(s):  
H Tomioka ◽  
Y Iwamura ◽  
Y Suzuki ◽  
S Ohtomo ◽  
Y Hashimoto
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Gamma Globulin ◽  
Protective Effects ◽  
Protective Activity ◽  
E Coli ◽  
Experimental Infections ◽  
Human Gamma Globulin

S-sulfonated gamma globulin (GGS), newly developed as a safe drug for intravenous use, was studied for its protective effects against some experimental infections in mice. Gamma globulin showed a good protective activity against infections due to Streptococcus pneumoniae and Escherichia coli and was moderately active against infections due to Staphylococcus aureus and Pseudomonas aeruginosa. In most cases, the potency of GGS was almost the same as that of original native gamma globulin. The duration of GGS activity in vivo was found to be comparable to that of native gamma globulin and much higher than that of pepsin-digested gamma globulin. In the control of infection due to E. coli, specific antibody was found to play a central role in the antibacterial action of GGS. When GGS was administered in combination with the antibiotics gentamicin and cefazolin for the control of infections due to S. pneumoniae or E. coli, a clear synergistic effect was observed.

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