Advanced Methods in Cellular Immunology: 1st Edition (Paperback) book cover

Advanced Methods in Cellular Immunology

1st Edition

By Rafael Fernandez-Botran, Vaclav Vetvicka

CRC Press

192 pages

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Paperback: 9780849321252
pub: 2000-05-26
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Description

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.

Table of Contents

SOURCES

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.

Subject Categories

BISAC Subject Codes/Headings:
MED044000
MEDICAL / Immunology
SCI017000
SCIENCE / Life Sciences / Cytology