Cellular Immunology

?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-a 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