Isolation Characterization, and Utilization of T Lymphocyte Clones - 1st Edition - ISBN: 9780122499203, 9780323145374

Isolation Characterization, and Utilization of T Lymphocyte Clones

1st Edition

Editors: C Fathman
eBook ISBN: 9780323145374
Imprint: Academic Press
Published Date: 28th January 1982
Page Count: 580
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Isolation, Characterization, and Utilization of T Lymphocyte Clones is a summary of information regarding T lymphocyte clones, including their usefulness. Organized into nine parts, the book begins with discussions on the soluble factors that can influence the growth of cloned T cells and the utilization of T cell hybridomas for analysis of T cell functions, emphasizing the biochemical and functional properties of helper and suppressor factors. The book then looks into the analysis of T cell clones and hybridomas using techniques of somatic cell genetics. The clonal analysis by limiting dilution, the characteristics of murine T cell clones reactive with alloantigens and soluble antigens, and the human T cell clones are described as well. This volume is valuable to those interested in the field of cloning of immunocompetent T cells.

Table of Contents



1 Introduction and Historical Overview



2 Differentiation within the Immune System: The Importance of Cloning

I. Comparison with the Nervous System

II. Cloning Methods

III. Questions of the Day


I. IL-1 (Lymphocyte Activating Factor) and IL-2 (T Cell Growth Factor)

3 Biochemical Characterization of Interleukin-2 (T Cell Growth Factor)

I. IL-2 Microassay

II. Purification of Human and Murine IL-2

III. Temperature, Chemical, and Enzymatic Treatments of IL-2

IV. Improved Cellular Sources for IL-2 Production and Characterization

V. Polyacrylamide Gel Electrophoresis of IL-2

VI. Molecular Prospects for the Future

VII. Conclusions


4 Production and Assay of Interleukin-1 (IL-1)

I. Production of IL-1

II. IL-1 Assay

III. Conclusions


5 Production and Properties of Human IL-2

I. Introduction

II. Culture Conditions for Generation of Human IL-2

III. Testing of Culture Supernatant for IL-2 Content

IV. Removal of PHA from IL-2-Containing Culture Supernatants

V. Long-Term Growth of Cytotoxic T Lymphocytes in IL-2

VI. Use of IL-2-Containing Preparations for Activation of Cytolytic Lymphocytes: IL-2 Providing the Proliferative Stimulus Necessary for Allogeneic IVS

VII. Conclusions


6 Signal Requirement for T Lymphocyte Activation

I. T Lymphocyte Propagation

II. T Lymphocyte Activation


II. T Cell Hybridomas and Their Products

7 An Overview

I. A Catalog

II. Problems

III. Lessons

IV. Conclusions


8 Factors, Receptors, and Their Ligands: Studies with H-2 Restricted Helper Hybridoma Clones

I. la-Associated Antigen Complex

II. Isolation of T Hybridomas

III. Characterization of the Hybridoma Clones

IV. Analysis of the Altered Self-Dual Recognition Problem

V. Isolation of Helper Factors


9 Properties of Antigen-Specific H-2 Restricted T Cell Hybridomas

I. Factor Production by T Cell Hybridomas

II. Use of T Cell Hybridomas to Examine Properties of the T Cell Receptor

III. T Cell Hybridomas Do Not Express Unexpected H-2 Spécificités

IV. Karyotypic Analysis of T Cell Hybridomas


10 Studies on an Antigen-Specific Suppressor Factor Produced by a T Hybrid Line

I. Derivation and Specificity of the Al Supressor Line

II B Cells as the Targets for Al Factor

III. Genetic Control of Suppression

IV. Interaction between Suppressor Factor and Monoclonal Antibodies in Regulation of the Immune Response

V. Conclusion


11 Characterization of Antigen-Specific Suppressor Factors from T Cell Hybridomas

I. Experimental Systems Used to Identify Suppressor T Cells and Their Products

II. Characterization of GAT- and GT-Specific TsF

III. Comparison of GAT-TsF and GT-TsF to Other Suppressor T Cell Factors


12 Suppression of Antibody Responses by a T Cell Hybridoma-Derived Haplotype-Specific Suppressor Factor

I. Properties of TsF-H

II. Mechanism of Action of TsF-H

III. Conclusions


13 Soluble Immune Response Suppressor (SIRS) Derived from T Cell Hybridomas

I. Characteristics of SIRS

II. Speculations


III. The Somatic Cell Genetic Analysis of Cytolytic T Lymphocyte Functions

14 An Overview

I. CTL Lines

II. Mutants

III. Somatic Cell Hybrids

IV. Outlook


15 Karyotype Evolution of Cytolytic T Cell Lines

I. Karyotype Analysis of Functional T Cell Lines

II. Discussion


16 Growth Regulation of Cytolytic T Cell Lines by Interleukin-2

I. Mechanisms of Action of Growth Factors

II. Mechanisms of Action of TCGF on a Cloned CTL Line


17 Correlation between Cytolytic Activity, Growth Factor Dependence, and Lectin Resistance in Cytolytic T Cell Hybrids

I. Introduction

II. Origin of CTL Hybrids

III. CTL Activity, VV Resistance, and CS Dependence of Hybrids

IV. Selection of C S - Variants from Cloned C S + Cytolytic Hybrids

V. Conclusions


IV. Clonal Analysis by Limiting Dilution

18 An Overview

I. Theory of Limiting Dilution

II. Experimental Conditions Required for Limiting Dilution Analysis of CTL-P Frequencies

III. The CTL-P Frequency Problem


19 Clonal Analysis of Helper and Cytolytic T Cells: Multiple, Independently Regulated

Precursor Sets at Frequencies Suggesting a Limited Repertoire

I. The Assay

II. Multiple Populations of T Precursor Cells

III. Independent Regulation of Each Precursor Population

IV. Lyt-Phenotypic Differences between T Precursor Populations

V. Conclusions


20 Frequency, Regulation, and H-2 Epitope Specificity of Alloreactive and H-2 Restricted

CTL Clones

I. Topographic Arrangement of Alloantigenic Determinants on the H-2Kk Molecule

II. Distinct CTL Subpopulations with Different Precursor Frequencies Detected by Limiting Dilution Analysis

III. Target Inhibition by mc Anti-H-2 of CTL Clones Generated in the Limiting Dilution System

IV. Target Inhibition of H-2Kk-Restricted, TNP-Specific CTL Clones



21 Production of Lymphokines by Murine T Cells Grown in Limiting Dilution and Long-Term Cultures

I. Introduction

II. Analysis of Lymphokine Release in Limiting Dilution Microcultures

III. Analysis of Lymphokine Production from Long-Term T Cell Clones and Lines

IV. Conclusions


V. Murine T Cell Clones Reactive with Alloantigens

22 An Overview

I. Conditions for Deriving T Cell Clones

II. Cytolytic T Cell Clones

III. Noncytolytic T Cell Clones

IV. Factors Produced by Alloreactive T Cell Clones

V. Conclusions


23 Cloned Continuous Lines of H-2 -Restricted Influenza Virus-Specific CTL: Probes of T

Lymphocyte Specificity and Heterogeneity

I. Properties of Influenza-Specific CTL Clones

II. Conclusions


24 Cytolytic T Lymphocyte Clones Recognizing Murine Sarcoma Virus-Induced Tumor


I. Optimal Microculture Conditions for the Generation of MoLV-Specific CTL Clones

II. Frequency Determination of MoLV-Specific CTL Precursors

III. Isolation and Maintenance of MoLV-Specific CTL Clones

IV. Specificity of MoLV-Specific CTL Clones

V. Surface Phenotype of MoLV-Specific CTL Clones

VI. Conclusions


25 The Specificity Repertoire of Cytolytic T Lymphocytes

I. Analysis of Receptor Specificity

II. CTL Receptor Repertoire of the B10.D2 Anti-H-2Kb Response

III. Specificity Repertoire of the bmll Anti-H-2Kb Response

IV. The C57BL/6 Anti-bmll CTL Response: A Model for Determinant Recognition

V. The Influence of MHC on Receptor Repertoire


26 Alloreactive T Cell Clones Which Recognize Hybrid Determinants



27 Anti-H-2 Reactivity of Mis-Specific T Cell Clones

I. H-2 Specificity of Uncloned Mis-Reactive T Cell Lines

II. Anti-H-2 and Anti-Mis Reactivity of T Cell Clones

III. Characteristics of Cloned Mis-Reactive T Cell Lines

IV. Conclusions


28 Lymphokine Production by Cytolytic and Noncytolytic Alloreactive T Cell Clones

I. Analysis of Lymphokine Production by a Noncytolytic T Cell Clone and Its Variant upon Alloantigenic Stimulation

II. Analysis of Lymphokine Production by Noncytolytic and Cytolytic Alloreactive T Cell Clones after Stimulation with Mitogen

III. Biological Separation of Lymphokine Activities Using Different T Cell Clones

IV. Heterogeneity of Cell Clones Producing Lymphokines

V. Conclusions


VI. Murine T Cell Clones Reactive with Soluble Antigens

29 An Overview



30 An Analysis of T Cell Antigen Recognition Utilizing T Cell Clones

I. Experimental Design

II. T Cell Antigen Specificity

III. Conformational Requirements of T Cell Antigen Recognition

IV. Antigen-Presenting Cell Function

V. Ia Restriction and Ir Gene Function

VI. Conclusions


31 Alloreactivity of Antigen-Specific T Cell Clones



32 Mechanism of B Cell Activation by Monoclonal T Helper Cell Populations

I. KLH-Specific Cloned T Cells Provide H-2 Restricted Help for Antibody Responses to TNP-KLH

II. Cloned Th Cells Can Be H-2 Restricted in Their Recognition of Accessory Cells but Not B Cells

III. After Specific Activation T Cell Clones Can Provide Antigen-Nonspecific Helper Activity

IV. Cloned Th Cell Supernatant Mediates Antigen- Nonspecific Help

V. Cloned Th Cells Can Function through a Pathway Requiring Lyb-5 + B Cells

VI. The Same Cloned Th Cells Function through Pathways Which Are either Restricted or Unrestricted for T h Cell Recognition of B Cell MHC Determinants 391

VII. The Same Cloned Th Cells Function through Pathways Which Activate Different B Cell Subpopulations

VIII. Conclusions


33 T Cell Lines and T Cell Clones Bearing Cross- Reactive Idiotype

I. Functional Characterization of the Helper Activity of Cell Line L.14

II. L.14 Cells Expressing Idiotypic-Like Determinants

III. Effect of Anti-Idiotypic Serum B658 on the Activity of Clone C.14.14

IV. Conclusions


34 Specific Regulation of Immune Responses by Products of T Cell Clones



VII. Human T Cell Clones

35 Cloning of T Lymphocytes in Man

I. PLT-Reactive Clones

II. Cytotoxic (CTL) Clones

III. Conclusions


36 Human T Cell Clones: Function, Specificity, and Cell Surface Markers

I. T Cell Clones Obtained Following Sensitization in Vitro

II. T Cell Clones following in Vivo Immunization

III. Surface Markers of Human T Cell Clones: Monoclonal Antibodies (mAB) Interacting with the Cytolytic Function

IV. Conclusions


37 Human T Cell Clones Reactive with Soluble Antigens: Methodology, Specificity, and MHC Restriction

I. Antigen Specificity of Soft Agar Colonies of Human T Cells

II. Genetic Requirements for Antigen Stimulation


VIII. Future Perspectives in the Utilization of T Cell Clones

38 Potential Use of Expanded T Lymphoid Cells and T Cell Clones for the Immunotherapy of Cancer

I. Traffic in Vivo of Cells Grown in T Cell Growth Factor

II. Effect in Vivo of Adoptively Transferred Cells Expanded in T Cell Growth Factor

III. Isolation of T Lymphocyte Clones Specifically Reactive with Tumor

IV. The Lysis of Fresh Syngeneic or Autologous Tumor Cells by Lymphoid Cells Expanded in T Cell Growth Factor

V. Conclusions


39 Cloned T Cells as a Tool for Molecular Geneticists: Approaches to Cloning Genes Which Encode T Cell Antigen Receptors

I. Immunoglobulin Expression in Cloned T Cells

II. Cloning Nonimmunoglobulin Genes from T Cells

III. Conclusions


IX. Appendix

I. Factors which Influence the Growth of T Lymphocyte Clones

A. Assay for Interleukin-1 (IL-1)

B. T Cell Lymphoma Model for the Analysis of Interleukin-1 (IL-1)

C. Preparation of TCGF from Rat Spleen Cells

D. Preparation of TCGF from a T Cell Tumor (EL-4)

E. Production of Human TCGF

F. Preparation of TCGF from Human Spleen Cells

G. IL-2 Microassay

H. Purification and Characterization of IL-2

II. Cell Fusion as a Technique for Generation of T Cell Clones

A. Production of Antigen-Specific, H-2 Restricted T Cell Hybridomas

III. Clonal Analysis by Limiting Dilution

A. Measurement and Calculation of CTL-P Frequencies

B. Limit Dilution Analysis of Functional T Cell Precursors

C. Limit Dilution Analysis of T Cells Releasing Lymphokines

IV. Murine T Cell Clones Reactive with Alloantigens

A. Clones of Noncytolytic Alloreactive Murine T Cells

B. Cloning of Alloreactive Murine T Cells

C. Isolation of Virus-Specific CTL Clones

V. Murine T Cell Clones Reactive with Soluble Antigens

A. Immunization and Long-Term Culture of Murine Immune Lymph Node Cells

B. Cloning of Soluble Antigen-Reactive Murine T Inducer (Th) Cells

C. Soft Agar Technique for Obtaining Antigen-Specific T Cell Clones

VI. Human T Cell Clones Reactive with Alloantigens and Soluble Antigens

A. Cloning of Alloreactive Human T Cells

B. Cloning and Expansion of Human Alloreactive T Lymphocytes

C. Establishment of Colonies of Antigen-Reactive Human T Cells



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© Academic Press 1982
Academic Press
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About the Editor

C Fathman

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