Methods for Studying Mononuclear Phagocytes

1st Edition - December 28, 1981
Editor: Dolph Adams
Language: English
eBook ISBN:
9 7 8 - 0 - 3 2 3 - 1 4 0 6 9 - 0

Methods for Studying Mononuclear Phagocytes is a practical guide to the study of mononuclear phagocytes that brings together various well-established and useful methods for… Read more

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Methods for Studying Mononuclear Phagocytes is a practical guide to the study of mononuclear phagocytes that brings together various well-established and useful methods for examining these cells. The technical protocols have been made detailed, specific, practical, and inclusive of the necessary mystique for immediate and direct application in the laboratory. The book is divided into 11 parts arranged according to the sequence of steps that would generally be followed to study a given population of mononuclear phagocytes: (I) methods for obtaining and culturing populations of human and animal mononuclear phagocytes; (II) methods for separating populations of leukocytes to enrich or deplete their content of mononuclear phagocytes; (II) criteria and techniques for identifying mononuclear phagocytes; (IV) methods for quantifying the number of mononuclear phagocytes; (V) techniques for studying the morphology of mononuclear phagocytes; (VI) methods for quantifying the biochemical constituents of mononuclear phagocytes; (VII) methods of quantifying phagocytosis, pinocytosis, and chemotaxis; (VIII) methods for quantifying the secretory products of mononuclear phagocytes; (IX) procedures for quantifying the destruction of tumor cells and of microorganisms by mononuclear phagocytes; (X) methods for studying the cell biology of mononuclear phagocytes; and (XI) techniques for studying mononuclear phagocytes in vivo, including procedures for estimating their kinetics, accumulation, identification, and microbicidal properties.

Contributors

Preface

I. Obtaining and Culturing Mononuclear Phagocytes

1 The Culture of Mononuclear Phagocytes: A Brief Overview

2 Murine Mononuclear Phagocytes from Bone Marrow

3 Obtaining and Culturing Murine Monocytes

4 Obtaining and Culturing Human Monocytes

5 Isolation of Human Monocytes

6 Culture of Human Monocytes in Microplates and Enzymatic Assays for Following Their Maturation

7 Peritoneal Mononuclear Phagocytes from Small Animals

8 Obtaining and Culturing Human and Animal Alveolar Macrophages

9 Obtaining Human Mononuclear Phagocytes from Colostrum

10 Obtaining Adherent Cells from Spleen

11 Obtaining Kupffer Cells

12 Obtaining Mononuclear Phagocytes from Disaggregated Neoplasms

13 Obtaining Mononuclear Phagocytes from Granulomas

14 Teflon Film as a Substrate for the Culture of Mononuclear Phagocytes

15 Effects of Husbandry and Mouse Strains on Mononuclear Phagocytes

16 Endotoxin Contamination and in Vitro Monocyte-Macrophage Function: Methods of Detecting, Detoxifying, and Eliminating Endotoxin

17 Continuous Cell Lines with Properties of Mononuclear Phagocytes

II. Separation of Mononuclear Phagocytes for Enrichment or Depletion

18 Separation of Mononuclear Phagocytes for Enrichment and for Depletion: An Overview

19 Separation of Murine Macrophages by Adherence to Solid Substrates

20 Separation of Human Monocytes and Guinea Pig Macrophages by Density Gradients of Metrizamide

21 Separation of Murine Mononuclear Phagocytes by Density Gradients of Percoll

22 Separation of Human Monocytes by Density Gradients of Percoll

23 Separation of Murine Mononuclear Phagocytes by Velocity Sedimentation at Unit Gravity

24 Use of Lidocaine for Detachment of Adherent Mononuclear Phagocytes

25 Use of Sephadex Columns to Deplete Mononuclear Phagocytes

26 Depletion of Mononuclear Phagocytes: Pitfalls in the Use of Carbonyl Iron, Carrageenan, Silica, Trypan Blue, or Anti-Mononuclear Phagocytes Serum

III. Identification of Mononuclear Phagocytes

27 Identification of Mononuclear Phagocytes: Overview and Definitions

28 Characteristics of Mononuclear Phagocytes from Different Tissues

29 Fc and C3 Receptors

30 Identification of Mononuclear Phagocytes by Ingestion of Particulate Materials, such as Erythrocytes, Carbon, Zymosan, or Latex

31 Heteroantisera Raised against Mononuclear Phagocytes

32 Monoclonal Antibodies as Tools for the Study of Mononuclear Phagocytes

33 Antisera against Ia Antigens

IV. Quantitation of Number of Mononuclear Phagocytes

34 Quantitation of Adherent Mononuclear Phagocytes by Inverted Phase Microscopy

35 Quantitation of DNA in Mononuclear Phagocytes

36 Lowry and Bradford Assays for Protein

V. Morphology of Mononuclear Phagocytes

37 Use of Wrights Strain and Cytocentrifuge Preparations

38 Use of Phase Contrast Microscopy

39 Use of Peroxidase Stain by the Kaplow Method

40 Use of Nonspecific Esterase Stain

41 Histochemical Stains for Macrophages in Cell Smears and Tissue Sections: β-Galactosidase, Acid Phosphatase, Nonspecific Esterase, Succinic Dehydrogenase, and Cytochrome Oxidase

42 Use of Transmission Electron Microscopy

43 Preparative Techniques for Scanning Electron Microscopy

44 Use of Ultrastructural Histochemistry

VI. Biochemical Constituents of Mononuclear Phagocytes

45 Lysosomal Enzymes

46 Microsomal Heme Oxygenase

47 Histamine O-Methyltransferase

48 5'-Nucleotidase Assay

49 Alkaline Phosphodiesterase I

50 Quantitation of Leucine Aminopeptidase of Mononuclear Phagocytes

51 Hexose Monophosphate Shunt Activity and Oxygen Uptake

52 Secretion of Superoxide Anion

53 Release of Hydrogen Peroxide

VII. General Functions of Mononuclear Phagocytes

54 Antibody-Dependent and Antibody-Independent Phagocytosis

55 Pinocytic Rate Using Horseradish Peroxidase

56 Chemotaxis of Human and Murine Mononuclear Phagocytes

VIII. Secretion by Mononuclear Phagocytes

57 Secretory Functions of Mononuclear Phagocytes: Overview and Methods for Preparing Conditioned Supernatants

58 Characterization and Classification of Macrophage Proteinases and Proteinase Inhibitors

59 Growth of Macrophage on Collagen, Elastin, and Glycoprotein-Coated Plates as a Tool for Investigating Macrophage Proteinases

60 Neutral Proteases by 3H-Labeled Casein

61 Plasminogen Activators by Use of 3H-Labeled Casein Substrate

62 Elastinolytic Enzymes

63 Microtiter Assay for Antiviral Effects of Human and Murine Interferon Utilizing a Vertical Light Path Photometer for Quantitation

64 Endogenous Pyrogen

65 Prostaglandins

66 Quantitation of Selected Complement Components

67 Lysozyme

IX. Destruction by Mononuclear Phagocytes

68 Destruction of Listeria monocytogenes in Vitro

69 Ingestion and Destruction of Candida albicans

70 Quantitation of Destruction of Toxoplasma

71 Destruction of Rickettsiae

72 Destruction of Leishmania

73 Destruction of Viruses

74 Cytostasis of Tumor and Nontumor Cells

75 Cytolysis of Tumor Cells by Release of [3H] Thymidine

76 Quantification of Cytolysis of Neoplastic Cells by Release of Chromium-51

77 Assessment of Cytolysis of Tumor Cells by Release of [125I] Iododeoxyuridine

78 Antibody-Dependent Cellular Cytotoxicity (ADCC) of Erythroid and Tumor Cells

X. Mononuclear Phagocytes as Tools in Cell Biology

79 Overview: The Macrophage in Cell Biology

80 Biosynthetic Radiolabeling of Cellular and Secreted Proteins of Mononuclear Phagocytes

81 Extraction, Identification, and Quantitation of Lipids

82 Synthesis, Cellular Turnover, and Mass of Cholesterol

83 Solute Uptake and Membrane Transport by Mononuclear Phagocytes

84 Use of Lactoperoxidase for Labeling Membrane Proteins

85 Binding of Synthetic Chemotactic Peptides as a Model of Ligand-Receptor Interaction

XI. Mononuclear Phagocytes In Vivo

86 Isolation of Phagosomes from Mouse Peritoneal Macrophages

87 Study of Mononuclear Macrophages in Vivo: Overview

88 Quantitation of the Inflammatory Accumulation of Mononuclear Phagocytes in Vivo

89 Systemic Labeling of Mononuclear Phagocytes

90 Labeling of Mononuclear Phagocytes in Granulomas and Inflammatory Sites

91 Identification of Fc and Complement Receptors in Tissue Sections

92 Determination of Macrophage-Mediated Antibacterial Resistance

Supplement to Table of Contents

Index