Cellular Immunology LabFax

Contributors

Abbreviations


1. Laboratory Animals

Welfare of Laboratory Animals

Health Monitoring

Anesthesia and Analgesia

Pre-Anesthesia

Anesthesia

Analgesia

Animal Models in Immunology

Induced Animal Models

Spontaneous Animal Models

Transplantation Immunology

Graft versus Host Reaction

Immunization of Animals for Antibody Production

Polyclonal Antibody Production

Monoclonal Antibody Production

Antigen Purity and Dose

Adjuvants

Routes of Administration

A Standard Procedure for the Production of Monoclonal Antibodies

A Standard Procedure for the Production of Polyclonal Antibodies

Bleeding

References


2. Tissues and Cells of the Immune System

Primary Lymphoid Tissues

Thymus

Bursa of Fabricius

Mammalian B Lymphopoiesis

Secondary (Peripheral) Lymphoid Tissues

Lymph Nodes

Spleen

Mucosa-Associated Lymphoid Tissue (MALT)

Lymphocyte Recirculation and Homing

Cells of the Immune Response

Lymphocytes

Nonlymphoid Cells

References


3. Leukocyte Development

Hematopoiesis

Lymphocyte Development

B Cells

T Cells

Markers of Cellular Activation

Leukocyte Lifespans

Age Dependence of Lymphocyte Subpopulations

Methods of Studying Lymphocyte Development

References


4. Tissue and Cell Culture

General Information

Biosafety Cabinets

Other Required Equipment

Methods of Sterilization

Biosafety Levels

Tissue Culture Media

Media Formulations

Supplements and Media Modifications

Antibiotics

Organ Culture

Long-Term Bone Marrow Cultures

Thymic Organ Cultures

Splenic Fragment Cultures

Cell Isolation

Methods of Tissue Dissociation

Enumeration of Cell Number

Washing and Removal of Dead Cells

Cell Separation Techniques

Dyes Used in FACS Analysis and Cell Sorting

Cell Culture

Cell Density

Use of Feeder Cells

Inactivation of Cells/Cell Substitutes

Use of Culture Supernatants

Polyclonal Activators

Cell and Tissue Preservation

Fixation

Cryopreservation

References


5. Cell Lines and Hybridomas

Cell Lines

Viral- and Gene-Mediated Transformation

Stromal Cell-Dependent B-Cell and T-Cell Lines

Antigen-Specific T-Cell Lines

Murine Helper T-Cell Lines

Human Helper T-Cell Lines

B-Cell Hybridomas

General Aspects of Hybridoma Production

Myelomas

Normal Lymphocytes

Cell Fusion

Selection of Hybridomas

Screening

Culture of Hybridoma Clones

Application of Monoclonal Antibodies

T-Cell Hybridomas

T-Lymphoma Lines

Normal T Lymphocytes

Fusion and Selection

Screening

Cloning

Definition and Purpose

Procedures

Verification of Clonality

Cell-Line Suppliers

References


6. Assays of Cellular Activity

Measurement of Early Activation

Phagocytosis and Microbicidal Activity

Chemotaxis and Chemokinesis

Proliferation

Cytokine Production

Antibody Secretion

Cytotoxicity

Helper Function

TH Lymphokine Secretion

Supernatant Helper Function

Limiting Dilution

Apoptosis

References


7. Cell-Surface Antigens

Introduction

References


8. Antigens Receptors

Structure

General Features

Immunoglobulin Heavy Chains

Immunoglobulin Light Chains

Class Differences between Ig Molecules

T-Cell Receptor Chains

Antigen Binding

Genes Encoding Antigen Receptors

Organization

Ig Heavy Chain Locus

The Ig Light-Chain Gene Loci

The TCR Gene Loci

Rearrangement of Antigen-Receptor Genes

Joining Signals and the 12/23 Rule

Mechanisms of Joining

Imprecision and Nucleotide Addition at Coding Joints

Components of the Rearrangement Machinery

Order of Rearrangement

Allelic Exclusion

Generation of Diversity

Combinatorial and Junctional Diversity

Somatic Hypermutation of Ig Genes

Expression of Ig and TCR Genes

Transcriptional Regulation of Ig and TCR Genes

Isotype Switching in IgH Genes

The Expressed V-Region Repertoire of TCR and Ig

The Ig V-Region Repertoire

The TCR V-Region Repertoire

Assembly and Cell-Surface Transport of Antigen Receptors

Components of the TCR Complex

Assembly and Surface Transport of CD3/TCR Complexes

Components of the Membrane-Bound Ig Complex

Assembly and Surface Transport of Membrane-Bound Ig

References


9. Antigen Processing and Presentation

Genetics

The Antigens

Antigen-Presenting Cells

The Interdigitating Dendritic Cell

The Macrophage

The B Cell

Inhibitors of Antigen Processing

The Antigen-Processing Pathway

The Molecules

Conclusions

References


10. The Major Histocompatibility Complex

Molecules Encoded within the MHC

Structure of Class I and class II Molecules

Maps of H-2 and HLA Genes (and Other Species)

Nomenclature of H-2 and HLA Alleles and Haplotypes

Typing Methods

References


11. Soluble Immunoregulatory Molecules

Introduction

Cytokines

Interleukins

Tumor Necrosis Factor-α and Lymphotoxin

Transforming Growth Factors

Interferons

Colony-Stimulating Factors

Soluble Cytokine Antagonists

Soluble Adhesion Molecules

Other Soluble Immune Regulatory Molecules

Immunoglobulins

References


12. Cellular Activation

Ligands and Receptors Involved in Immune-Cell Activation

Cell Lines Useful for Studying Signal Transduction

Tyrosine Kinase Activation by Receptors on Cells of the Immune System

PTK Substrates

Tyrosine Kinase-Regulated Signal Transduction Pathways

The Phospholipase C Pathway

The Phosphatidylinositol 3-Kinase Pathway

The p21ras Pathway

The MAP Kinase Pathway

Activation of Phospholipase C by G Protein-Linked Receptors

Regulation of cAMP Levels

Phosphatidylcholine Hydrolysis

Glycosyl-Phosphatidylinositol (GPI) Hydrolysis

Methods Used in Studying Signal Transduction

References

Index