Human Hybridomas and Monoclonal Antibodies

I B-Cell Lines, Hybridomas, and Monoclonal Antibodies.- A. Background and General Strategies.- 1 Human Hybridomas and Monoclonal Antibodies: The Biology of Cell Fusion.- I. Introduction.- II. Historical Development of Hybridomas.- III. Problems with Conventional Immunization.- IV. Advantages of the Monoclonal Antibody Technology.- V. Difficulties and Technical Problems in the Production of Monoclonal Antibodies.- VI. Human Hybridomas and Monoclonal Antibodies.- VII. Methods for Making Human Monoclonal Antibodies.- VIII. Limitations in Making Human Monoclonal Antibodies.- IX. Enrichment for Sensitized Lymphocytes and in Vitro Stimulation of Lymphocytes.- X. Fusion Variables.- XI. Selection of Hybrids.- XII. Freezing and Thawing Hybridomas.- XIII. Chromosome Stability in Hybridomas.- References.- 2 Fusion Partners for Production of Human Monoclonal Antibodies.- I. Murine Plasmacytomas as Fusion Partners.- II. Human Plasmacytomas as Fusion Partners.- III. Human Lymphoblastoid Cell Lines as Fusion Partners.- IV. Lymphoblastoid versus Myeloma Cells as Fusion Partners.- V. Search for Better Fusion Partners.- References.- 3 Production of Human Monoclonal Antibodies Using Epstein-Barr Virus.- I. Introduction.- II. The Biological Properties of Epstein-Barr Virus.- III. Human Monoclonal Antibody Production.- IV. Future Perspectives.- References.- 4 The Epstein-Barr Virus-Hybridoma Technique.- I. Epstein-Barr Virus Technology.- II. The Epstein-Barr Virus-Hybridoma Technique.- III. Anti-Tetanus Monoclonals from in Vitro Immunized Cultures.- IV. Anti-Mycobacterium leprae Monoclonals.- V. Human Monoclonal Antibodies against Lung Carcinoma Cell Lines.- VI. Human Monoclonal Antibodies against Myelin-Associated Glycoprotein.- VII. New Fusion Partners.- VIII. Ascites Growth of Human Hybridomas.- IX. Epstein-Barr Virus-Hybridoma Protocol.- X. The Advantages of Human Monoclonal Antibodies.- XI. Possible Limitations.- XII. Conclusions.- References.- 5 Strategies for Stable Human Monoclonal Antibody Production: Construction of Heteromyelomas, in Vitro Sensitization, and Molecular Cloning of Human Immunoglobin Genes.- I. Introduction.- II. General Properties of Mouse-Human Heteromyeloma and Heteromyelolymphoma Cell Lines.- III. Optimal Antigen-Primed Human B-Lymphocyte Fusion Partners.- IV. Permanent Preservation of Immunoglobulin Production.- References.- B. Applications to Infectious Diseases.- 6 Production and Characterization of Human Monoclonal Antibodies against Gram-Negative Bacteria.- I. Introduction.- II. Human Hybridomas Secreting Antibody to Escherichia coli.- III. Human Monoclonal Antibody Production in Nude Mice.- IV. Characterization of Antibody Specificity.- V. Discussion.- VI. Summary.- References.- 7 Human Monoclonal Antibodies to Defined Antigens: Toward Clinical Applications.- I. Production of Monoclonal Antibodies for Diagnostic and Therapeutic Use.- II. Peripheral Blood as a Source of Antigen-Primed Lymphocytes.- III. A Comparison of Mouse and Human Lymphoid Fusion Partners.- IV. Human Antibodies to Specific Antigens.- V. Clinical Applications.- VI. Summary.- References.- 8 Production of Human Monoclonal Antibodies Using a Human—Mouse Fusion Partner.- I. Introduction.- II. Generation of a Human-Mouse Fusion Partner.- III. Human Anti-A Red Blood Cell Antibody.- IV. Human Monoclonal Antibodies to Varicella Zoster Virus.- V. An Anti-Epstein-Barr Virus IgM Human Monoclonal Antibody.- VI. Human Monoclonal Antibodies to Mycobacterium leprae.- VII. Concluding Comments.- References.- 9 In Vitro Expansion of Human B Cells for the Production of Human Monoclonal Antibodies.- I. Introduction.- II. Methods.- III. In Vitro B-Cell Expansion with EBV.- IV. Class Switching of Human Hybridomas.- V. In Vitro Antigen-Specific Expansion of B Cells.- VI. Conclusions.- References.- 10 Cell-Driven Viral Transformation.- I. Introduction.- II. Derivation of the Driving Cell Line.- III. Application of the Cell-Driven Viral Transformation Process to the Generation of Specific Human Monoclonal Antibodies.- IV. Discussion.- References.- C. Applications to Cancer.- 11 The Generation of Human Monoclonal Antibodies and Their Use in the Analysis of the Humoral Immune Response to Cancer.- I. Introduction.- II. Properties of Drug-Marked Fusion Partners.- III. Human Monoclonal Antibodies Directed against Cellular Antigens.- IV. Summary.- References.- 12 Design and Production of Human Monoclonal Antibodies to Human Cancers.- I. Introduction.- II. Development and Characterization of UC 729–6.- III. Enzyme Immunoassay to Qualitate and Quantitate Human Immunoglobulin.- IV. Use of Regional Draining Lymph Nodes of Cancer Patients as a Source of Antitumor Antibodies.- V. Detection of Cell-Bound Antigens.- VI. Concluding Remarks.- References.- 13 Human-Human Hybridoma Technology: Five Years of Technical Improvements, and Its Application in Cancer Biology.- I. Introduction.- II. Improvement of Malignant Fusion Partners.- III. In Vitro Antigen Priming.- IV. Fusion Procedures, Cultivation of Human-Human Hybrids, and Screening for Immunoglobulin Production.- V. Human Monoclonal Antibodies against Tumor-Associated Antigens.- VI Conclusion.- References.- D. Applications to Autoimmunity.- 14 The Production of Monoclonal Antibodies by Human-Human Hybridomas: Their Application to Studies of Autoimmune Diseases.- I. Introduction.- II. Preparation of Human-Human Hybridomas.- III. Technical Considerations.- IV. Monoclonal Human Lupus Autoantibodies.- References.- 15 Human Monoclonal Autoantibodies Reactive with Multiple Organs.- I. Introduction.- II. Establishment of Continuous Cell Lines Producing Autoantibodies.- III. Comparison of Methods for Generating Monoclonal Autoantibodies.- IV. Methods of Screening for Autoantibodies.- V. Evaluation of Tissue Reactivity of Monoclonal Autoantibodies.- VI. Purification of Human Autoantigens by Immunoaffinity Chromatography.- VII. Discussion.- References.- 16 Principles of in Vitro Immunization of Human B Lymphocytes.- I. Introduction.- II. Principles That Govern the Activation of B Lymphocytes. Helper Requirements.- III. Antigens.- IV. Antigen-Specific Induction of Antibody Synthesis in Culture of Human Lymphoid Cells.- V. The Role of Suppressor Cells in the Generation of Antibody-Forming Cells in Vitro.- VI. Measures to Amplify Antibody Production.- VII. In Vitro Activation of B Cells Applied to the Production of Human Monoclonal Antibody.- References.- E. Special Topics.- 17 Human-Human Hybridomas in the Study of Immunodeficiencies.- I. Introduction.- II. Immunodeficiency Patients Used in Hybridoma Formation.- III. Lymphocyte Activation and Hybridization.- IV. Characterization of the Hybridomas Obtained from the Peripheral Blood B Cells of Immunodeficiency Patients.- V. Immunoglobulin Gene Analysis of the Hybridomas Derived from Immunodeficiency Patients.- VI. RNA Processing of µ Heavy Chain Message in the Hybridomas Derived from Immunodeficiency Patients.- VII. Discussion.- References.- II Human T-T Hybridomas.- 18 Human T-Lymphocyte Subsets and T-T Hybridomas: An Overview.- I. Introduction.- II. Functional T-Cell Subsets in Man.- III. The Role of Lymphokines in Immunoregulation.- IV. Human T-T Hybridomas.- References.- 19 Production of Human T-Cell Hybridomas by Electrofusion.- I. Introduction.- II. Fusion Partners in Polyethylene Glycol Fusion and Electrofusion.- III. Polyethylene Glycol Fusion.- IV. Electrofusion.- V. Characterization of Fusion Products Obtained by Polyethylene Glycol and Electrofusion.- VI. Fusion Protocols.- VII. Discussion.- References.- 20 Human B-Cell Growth and Differentiation Factors Secreted by T-Cell Hybridomas.- I. Introduction.- II. Assay Systems for Human B-Cell Proliferation and Differentiation.- III. Hybridization of Mitogen-Activated Human Peripheral Blood T Lymphocytes.- IV. Development of a Human T-Cell Hybridoma Secreting B-Cell Growth Factor.- V. Development of a Human T-Cell Hybridoma Secreting Both B-Cell Growth Factor and B-Cell Differentiation Factor.- VI. Application of T-Cell Hybridoma-Derived B-Cell Growth Factor and B-Cell Differentiation Factor to the Study of Human B-Cell Immunoregulation.- VII. Summary and Conclusions.- References.- 21 Generation and Characterization of Human T-Cell Hybridomas That Constitutively Produce Immune Interferon.- I. Introduction.- II. Isolation of a Parent Cell Line Deficient in Hypoxanthine Phosphoribosyltransferase.- III. Derivation and Characterization of Human T-Cell Hybridomas.- IV. Production of Immune Interferon by Hybridomas.- V. Application of Hybridomas to Structure-Function Studies of Immune Interferon.- VI, Summary.- References.- 22 Selection of Human T-Cell Hybridomas That Produce Inflammatory Lymphokines by the Emetine-Actinomycin D Method.- I. Introduction.- II. Emetine-Actinomycin D Selection Method.- III. Migration-Inhibitory Factor and Macrophage-Activating Factor Activities in Culture Supernatants of Hybridomas.- IV. Functional Stability of Human T-Cell Hybridomas.- References.- 23 Human T-T Hybridomas Specific for Epstein-Barr Virus: Generation and Function.- I. Introduction.- II. Choice and Construction of the Malignant T Lymphoblast.- III. Selection and Preparation of Epstein-Barr Virus-Specific Normal T-Cell Parent.- IV. Hybridization.- V. Proof of Hybridization.- VI. Testing for Antigen Specificity.- VII. HLA-DR Restriction of Recognition.- VIII. General Considerations.- IX. Summary.- References.- 24 Factors Generated by Human T-Cell Hybridomas Regulate B-Cell Activation, Polyclonal Differentiation, and Isotype Expression.- I. Introduction.- II. Methodology for Human T-Cell Hybridomas.- III. Factor Secretion by Human T-Cell Hybridomas.- IV. Model of B-Cell Maturation.- References.- 1. Human T- and B-Cell Lines.- 2. Preparation of Mutant Cell Lines.- 3. Freezing Human Cell Lines.- 4. Mycoplasma Testing.- 5. Peripheral Blood Lymphocyte Separation from Whole Blood or Buffy Coats.- 6. Separation of Human T and Non-T Lymphocytes from Peripheral Blood.- 7. Panning for Human T-Lymphocyte Subpopulations.- 8. Human Hybridoma Tube Fusion Protocol.- 9. Plate Fusion Technique for Nonadherent Cells.- 10. Identification of Human Chromosomes in Mouse-Human Lymphocyte Hybrids.- 11. Epstein-Barr Virus Transformation.- 12. Growth of Human Cell Lines in Mice.- 13. Methods of Large-Scale Tissue Culture.- 16. Soft Agar Cloning Protocol.- 17. Cloning by Limiting Dilution.- 18. Reverse Plaque Assay.- 19. Assay of Interleukin 1 (IL-1).- 20. Assay of Interleukin 2 (IL-2).- 21. Assay of B-Cell Growth and Differentiation Factors.- 22. Indirect Immunoglobulin ELISA Protocol.- 23. Purification of Human Immunoglobulins.- 24. A Nitrocellulose Strip Method for Isotyping Monoclonal Antibodies.- 25. Immunoprecipitation of Antigens Using Polystyrene Balls.- 26. Immunostaining Using Monoclonal Antibodies.- 27. Immunoblotting.- Manufacturers and Distributors.