Antigen-Antibody Reactions In Vivo

Antigen-Antibody Reactions In Vivo

Methods in Immunology and Immunochemistry, Vol. 5

1st Edition - December 28, 1976

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  • Editors: Curtis A. Williams, Merrill W. Chase
  • eBook ISBN: 9781483220604

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Methods in Immunology and Immunochemistry, Volume V: Antigen-Antibody Reactions In Vivo deals primarily with immune phenomena in tissues or in cell preparations. This book covers a variety of topics, including anaphylaxis, tolerance, immune suppression with chemical agents, radiation effects, antibody synthesis in vitro, immunological methods, and applied electron microscopy. Organized into 10 chapters, this volume begins with an overview of systemic anaphylaxis investigations in other more resistant species. This text then presents the analysis of mechanisms involved in the pathogenesis of the Arthus phenomenon, which shed light on the understanding of other lesions of hypersensitivity. Other chapters consider the effects of antigen–antibody interaction on connective tissue. This book discusses as well the degree and duration of acquired tolerance. The final chapter deals with the application of electron microscopy in the elucidation of the mechanisms of immune reactions. This book is a valuable resource for immunologists, students, and research workers.

Table of Contents

  • Contributors to Volume


    Contents of Other Volumes

    Chapter 19. Anaphylaxis

    A. Systemic Anaphylaxis

    Active Anaphylaxis: Antigens

    Procedures with Guinea Pigs







    Passive Anaphylaxis: Principles

    Procedures with Various Species

    B. Local Anaphylaxis

    Active Cutaneous Anaphylaxis

    Passive Cutaneous Anaphylaxis

    General Procedures

    Special Procedures for PCA in Guinea Pigs

    Procedures for Mice, Rats, Rabbits

    Semiquantitative Passive Cutaneous Anaphylaxis

    Prausnitz-Küstner Skin Tests in Man for Reagins (IgE Antibody)

    Passive Transfer of Human Reagins into the Monkey

    C. Anaphylaxis in Isolated Tissues and Cells

    1. Anaphylaxis in Isolated Tissues

    2. Anaphylactic Responses with Isolated Lung Slices

    3. Anaphylactic Release of Histamine from Rat Mast Cells

    4. Responses of Isolated Leukocytes and Platelets in Reaginic Allergy

    D. Assay of Pharmacologically Active Materials

    1. Assay of Histamine

    2. Assay of Serotonin

    3. Biological Assay of Slow Reacting Substances—SRS-A, Bradykinin, Prostaglandins,

    4. Immunoassay for Bradykinin and Kininogen

    Chapter 20. The Arthus and Related Reactions

    A. Cutaneous Arthus Reactions

    Active, Direct Passive, Reversed Passive Arthus Reactions

    Local Passive Arthus Reactions

    Reactions Induced by Soluble Complexes

    Tissues Employed in Arthus Studies

    Elicitation Requirement for Precipitating Antibody and for Complement Fixation

    B. Methods of Modifying Arthus Reactions

    Elimination of Polymorphonuclear Leukocytes

    Depletion of Serum Complement

    Preparation of Cobra Venom Factor



    C. Arthus Phenomenon in Disease Processes

    Serum Sickness Arteritis

    Acute Nephrotoxic Nephritis

    Cutaneous Reaction to Anti-Kidney Basement Membrane Antibody

    Chapter 21. Analysis of Antigen-Antibody Reactions in Cornea


    Local Antigen-Antibody Reactions in the Cornea

    Wessely Phenomenon

    Anaphylactic Keratitis

    Local Intracorneal

    Injections in Rabbits with Both Antigen and Antibody

    Chapter 22. Immunological Tolerance

    A. Introduction

    B. Tolerance to Heterologous Serum Proteins and Related Substances

    1. Induction in Neonatal Animals, 188. Rabbits

    2. Induction of Tolerance in Normal Adult Animals

    3. Induction of Tolerance in Irradiated Normal Animals

    C. Tolerance to Homologous and Heterologous Erythrocytes




    D. Induction of Tolerance to Chemical Haptens

    Induction by Feeding

    By Injection

    E. Induction of Tolerance in Vitro


    Polymerized Flagellin


    Chapter 23. Immune Suppression and Induction of Tolerance with Chemical Agents

    A. Introduction

    B. Biochemical and Pharmacological Properties of the Agent



    5-Fluoro-2-Deoxyuridine (FUDR)


    Actinomycin D

    C. Immunosuppressive Properties of the Agents

    1. 6-Mercaptopurine

    2. Cyclophosphamide

    3. Methotrexate

    4. 5-Fluoro-2-deoxyuridine (FUDR)

    5. Actinomycin D

    Chapter 24. Methods of Applications of Radiation in Immunological Research

    A. Radiation Measurements

    Characteristics of the X-Ray Beam


    Roentgen Units

    Importance of Rate of Exposure

    Problems in Determining Absorbed Dose

    Electronic Equilibrium

    B. Representative Effects of Radiation

    1. In Vitro Radiation Effects on Serum

    2. Total Body Irradiation

    3. Low Level Protracted Semicontinuous Radiation

    4. Partial Body Radiation: Selective Shielding

    C. Concluding Remarks


    Depression or Enhancement of Antibody Synthesis by X-Ray Irradiation

    Chimeras Produced by Irradiation

    Late Effects of Radiation

    Chapter 25. Phagocytosis

    A. Phagocytosis

    1. Preparation of Phagocytic Cells

    2. Media and Cell Counting

    3. Direct Evaluation of Ingestion

    4. Evaluation of Phagocytosis by Measuring Phagocytic Metabolism

    B. Clearance in Vivo


    Antigen-Antibody Complexes

    Bacterial Clearance

    Clearance of Carbon from the Blood Stream

    Effect of RES Stimulation

    Clearance of Bacteria from Serous Cavities

    C. Elimination Of 51Cr-Labeled Erythrocytes


    Studies of Normal Erythrocyte Lifespan

    Assessment of Anti-Erythrocyte Antibodies

    Patterns of Elimination

    Labeling and Sampling Methods


    Chromium Dose Calculation

    Blood Collections

    Elimination Rates

    Immediate Elimination by Natural Antibodies or by RES

    Immune Clearance

    Onset of Antibody Formation, Slow Immune Clearance, Calculations

    Sensitivity of the 51Cr-Elimination Method

    Chapter 26. Antibody Synthesis in Vitro

    A. Tissue Culture Methods for Antibody Production in Vitro

    1. Introduction

    2. Technique of Lymph Node Cultures

    3. Culture of Spleen Cells for Primary Response

    4. Measurement of Synthesized Antibody

    B. Feeder Cell Techniques For Antibody Formation in Vitro

    General Considerations

    Immunization by Diphtheria Toxoid in Rats in Rabbits

    Stimulation of Immune Rat Lymph Node Fragments by Diffusible Materials Produced by Nurse Cells in a Parabiotic Chamber

    Stimulation of Immune Rabbit Lymph Node Fragments by Rabbit Thymus Preparations in Miniature Parabiotic Chamber, or by Thymosin

    C. Single-Cell Techniques in Antibody Production


    Micromanipulation Equipment

    Advance Preparation of the Cover Slip in the Micromanipulation Chamber

    Cell Depot Droplets for Selection of Single Cells

    Handling of Mammalian Cells

    Isolation and Washing Procedures

    Single Cell Isolation

    Special Bicarbonate-Free Nutrient Medium

    Detection of Antibody to Bacterial Antigen Synthesized by Single Cells

    Bacterial Adherence to Single Cells

    Immobilization of Motile Bacteria

    Observations on Bacterial H and O Agglutination

    Quantitation of Antibody in Microdroplets by Serial Dilution

    Determination of Mercaptoethanol Sensitivity of Antibody

    Handling of Plaque-Forming Cells (PFC)

    Liquid Monolayer Technique and Inversion of Micromanipulation System

    Carboxymethyl Cellulose Plaque Technique

    Serial Transfers of PFC

    Micromanipulation and Autoradiography of Cells Labeled in Vitro

    Chromosomal Analysis of Antibody-Forming Cells Arrested in Metaphase

    Deoxycholate Method of Rupturing Cells

    Microadaptations of the Ford Technique

    Microradioiodination of PFC-Secretions

    Electrophoresis on Cellulose Acetate


    Microdensitometric Analysis

    D. Plaque Techniques for Recognizing Individual Antibodyforming Cells

    1. Standard Techniques

    2. Indirect Techniques

    3. The Thin Layer Technique

    4. Modified Techniques

    5. Hemolytic Plaques to Detect Synthesis of Antibodies to Diverse Antigens

    6. Plating Error and Efficiency of Plating

    E. Rosette-Plaque Methods for Detection Of Allotype and Hapten Receptors In Plaque-Forming Cells


    Lymphocyte Receptor for Sheep Erythrocytes vs. Direct Plaque Technique

    Expression of Allotype Heterozygosity

    Detection of Individual Hapten Binding and Anti-Hapten Secreting Cells

    Preparation of Lymph Node Cells, Types of Target Cells, Rosette Plaque Assay

    Rosette Formation vs. Plaquing: Lymphocytes from NIP-Immunized Rabbits Passively Sensitized by Anti-Rabbit Allotype Antibody

    Chapter 27. Immunohistological Methods

    A. Preparation of Tissue for Study With Labeled Antibody and Light-Microscopy

    1. Introduction

    2. Theory and Application

    3. Preparative Methods

    B. Fluorescent Antibody as Specific Cytochemical Reagents

    1. Preparation of Fluorescein-Coupled Globulins

    2. Tissue Preparation for Immunofluorescence Staining

    3. "Staining" with Labeled Antibody

    4. Controls on Specificity of Fluorochrome Staining

    5. Microscopic Equipment for Observing Fluorescence

    6. Photography of Preparations Stained with Fluorescent Antibody

    7. Tetramethylrhodamine Isothiocyanate (TRITC) Conjugates

    8. Visualization of Antigens by Fluorochrome Reagents of Contrasting Color

    9. Staining with FITC-Coupled and Ferritin-Coupled Univalent Antibody Fragments ("Hybrid Antibodies")

    C. Fluorescent Proteins for the Determination of Cell Death


    Preparation of FITC-Labeled Bovine Albumin

    Detection of Dead Cells: in Organ Cultures

    In Tissues in Vivo

    Fixation and Sectioning of Tissues

    Differentiation of Phagocytosed FIBSA and Uptake by Dead Cells

    D. Staining of Histones in Nuclei of Lymphoid Cells

    Applicability: Lysine-Rich Histones in Smears, Frozen Cryostat Sections of Mammals, Aves, Plants, Squash Preparations, Electropherograms of Isolated Histones on Cellulose Acetate


    Formalin Fixation, Ammoniacal Silver, Formalin Developer

    Cytophotometry for Relative Density of Stain

    Selective Staining of: Normal Lymphoid Cells vs. Antigen-Stimulated Animals,

    Nuclear Histones of Cancer Cells

    Comments: Noninterference by Prior or Subsequent Feulgen Staining

    Characterization of Cell Types by Altered Prestaining with Formalin

    Staining of Arginine-Rich Histones by Fast Green after DNA-Extraction

    Chapter 28. Application of Electron Microscopy to Problems in Immunology

    A. Introduction

    Study of Cells during Antibody Synthesis, Distribution of Antigens, Cell-Specific Antigenic Markers. Cytopathology, Antibody as Cytologic Stain, Requirement for Electron-Dense Antibody Molecules

    Complexing of Antibody with Ferritin, with Enzymes, with Viruses

    B. Immune Labeling for Electron Microscopy

    1. Ferritin-Conjugated Antibody

    2. Hybrid Antibody Markers for Electron Microscopy

    3. Enzyme-Labeled Antibody Markers for Electron Microscopy

    C. Electron Microscopy of Immunoglobulins and Immune Complexes

    General Considerations for Visualizing Soluble Proteins by Negative Contrast,

    Support Films: Carbon Covered Nitrocellulose Layers or Adhesive-Glue Grids,

    Negative-Staining Heavy-Metal Salts

    Properties of Protein Molecules Capable of Visualization

    Staining Procedures

    Sequential Drop Method Allowing Reactions on Grids before Staining

    One-Step Direct Staining

    Carbon Film Transfer Method from Mica

    Immunological Applications, 502: Rabbit IgG Linked by Bivalent Hapten for Structures of IgG, Fc, Fab

    Complement Fixation by Antigen-Antibody Tetramers

    Human IgA Dimer

    IgM Ultrastructure

    Clq Ultrastructure

    C3 Component of Complement

    Author Index

    Subject Index

Product details

  • No. of pages: 586
  • Language: English
  • Copyright: © Academic Press 1976
  • Published: December 28, 1976
  • Imprint: Academic Press
  • eBook ISBN: 9781483220604

About the Editors

Curtis A. Williams

Merrill W. Chase

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