1st Edition
Advanced Methods in Cellular Immunology
Immunologists as well investigators in other disciplines may often use protocols involving the isolation, cultures and characterization of different types of leukocytes. Advanced Methods in Cellular Immunology is a collection of techniques in an easy-to-use format.
Each chapter provides readers with related program information, a step-by-step description of the methodology, alternative techniques, pertinent references, and information about commercial sources for materials and regents. In addition to leukocytes, the authors guide readers through the processes of cell culture as well as inflammation and autoimmunity in a variety of animal models.
Covering topics such as PCR and Apoptosis, this book will serve a guide to commonly used procedures in cellular immunology while utilizing both human and murine models.
Preparation of Cell Suspensions
Isolation of Cells
Mitomycin C Treatment
Preparation of Mycobacteria Suspension
Preparation of Listeria Suspension
ELISA
Incorporation of TritiatedThymidine into DNA
MTT Assay
Cr Labeling
ISOLATION AND CHARACTERIZATION OF NEUTROPHILS
Percoll Sedimentation and Ficoll-Paque Centrifugation
Magnetic Cell Sorting with Anti-CD15 Beads
Functional Assessment
ISOLATION AND CHARACTERIZATION OF EOSINOPHILS
Discontinuous Density Percoll Gradients
Magnetic Cell Sorting with Anti-CD16 Beads
Functional Assessment
ENRICHMENT OF HUMAN BASOPHILS
MACROPHAGES
Isolation of Macrophages
Activation of Macrophages
Production of Oxidative Metabolism Intermediates
Phagocytosis
Killing of Bacteria
Killing of Bacteria - Colorimetric Evaluation
Microbicidal Assay
B LYMPHOCYTE CLONING
Murine B Lymphocyte Cloning Assay
Proliferation of Pre-B Cell Colonies in Methycellulose
Clonal Assay for Murine Lymphohematopoietic Progenitors
Clonal Assay for Fetal Cells
Long-Term Bone Marrow Cultures
CELL CULTIVATION IN-VITRO
Tissue Culture Laboratory
Aseptic Techniques
Safety Concerns
Substrate
Basic Principles
Primary Cultures
Freezing of Cell Lines
ANALYSIS OF CYTOKINE OR CYTOKINE RECEPTOR mRNA
Introduction
Total RNA Isolation
Reverse Transcription-Polymerase Chain Reaction (RT-PCR)
INTRACELLULAR CYTOKINE STAINING
Cell Activation
Staining
CELL CYTOTOXICITY
Preparation of Mouse Cytotoxic T Cells
Whole Blood Assay
DNA Fragmentation
Apoptosis
Quantitation of Apoptosis Using Fluorescent Dyes
Quantitation of DNA Fragmentation
Target Cell Detachment Assay
Cytolysin Hemolytic Activity
Isolation of Cytoplasmic Granules
Esterase Assay
Activation of Antibodies
Activation by Phorbol Myristate Acetate and Ionophore
MIXED LYMPHOCYTE REACTION
Unidirectional MLR
ANIMAL MODELS OF AUTOIMMUNE DISEASES
Diabetes Mellitus
Nonobese Mice
Chemically Induced Diabetes
Experimental Autoimmune Encephalomyelitis
Rheumatoid Arthritis
BUFFERS AND MEDIA
COMMERCIAL SOURCES
INDEX.
Biography
Rafael Fernandez-Botran, Vaclav Vetvicka
