Tumour immunobiology : a practical approach :

목차 (Table of Contents)

• CONTENTS
• List of contributors = xxiv
• List of abbreviations = xxix
• 1. Preparation of tumour cell-lines / A. P Wilson = 1
• 1. Introduction = 1
• 2. Materials required for the preparation of tumour cell-lines = 2
• 3. Collection of clinical material = 4
• 4. Disaggregation of solid tumours = 4
• Mechanical disaggregation of tumours = 5
• Disaggregation of tumours by enzyme digesstion = 6
• 5. Effusions = 8
• 6. Clean-up procedures = 8
• Removal of red blood cells and dcad cells = 8
• Preliminary separation of epithelial and stromal cells = 9
• 7. Setting up primary cultures = 9
• 8. Characterization = 11
• 9. Pitfalls and trouble-shooting = 13
• Cross-line contamination = 13
• Mycoplasma contamination = 13
• Genotypic and phenotypic drift = 13
• Poor success rate for establishing lines = 14
• References = 14
• 2. Assessment of the immunogenicity of tumour models / Fred R. Miller ; Bonnie E. Miller ; Gloria H. Heppner = 17
• 1. Introduction = 17
• 2. Immunogenicity and antigenicity = 18
• 3. Statistical considerations in assessment of immunity = 19
• 4. Tumour cell source for immunization or challenge = 20
• 5. Host irradiation = 24
• 6. Elimination of suppressor cells = 25
• 7. Summary = 25
• Acknowledgements = 25
• References = 25
• 3. Flow cytometry of lymphocytes and tumour cells / John Lawry = 27
• 1. Introduction = 27
• 2. Sample considerations = 30
• Natural sources of cell suspensions = 30
• Solid tumours = 34
• Sample storage and fixation = 41
• 3. Standardization = 45
• The choice of controls = 45
• Alignment and calibration = 46
• 4. Current applications of flow cytometry = 47
• Haematogenous and lymphoid tumours = 47
• Clinical applications of the analysis of solid tumours = 49
• Further reading = 55
• Acknowledgements = 56
• References = 56
• 4. (Glyco-) protein antigens and peptide epitopes of tumours / Michael R. Price = 59
• 1.Introduction = 59
• 2.The tumour cell surface = 60
• 3. Anti-tumour antibodies = 61
• Antibody storage and stability = 61
• Antibody purification = 61
• 4. Immunoassays = 63
• Radioisotopic antig1obulin assay = 64
• Competitive inhibition of antibody binding = 65
• Sandwich or double-determinant immunoassay = 66
• 5. Tumour homogenization and sub-cellular fractionation = 68
• Homogenization = 68
• Sub-cellular fractionation = 69
• 6. Antigen degradation studies = 69
• 7. Antigen solubilization = 70
• 8. Antigen purification = 70
• Strategies = 70
• Lectin affinity chromatography of glycoproteins = 71
• Antibody affinity chromatography = 71
• 9. Radiolabelling cell-surface proteins and glycoproteins = 72
• 10. Imlnunoprecipitation = 73
• 11. Immunoblotting = 75
• 12. Epitope mappillg = 75
• 13. Conclusions = 77
• Acknowledgement = 77
• References = 77
• 5. Isolation and characterization of human natural killer cells / Tuomo Timonen ; Anna Maenpaa ; Panu Koanen (with additional material by U. P. Thorgeirsson and A. R. Mackay) = 81
• 1. Introduction = 81
• 2. Characteristics of NK cells = 82
• Morphology and physical characteristics = 82
• Surface phenotype = 82
• Regulation of NK activity = 84
• 3. Preparation of peripheral blood mononuclear cells (PBMC) = 84
• 4. Purification of human natural killer cells = 84
• Emichment of large granular Iymphocytes from peripheral blood = 84
• Depletion of non-NK cells from LGL-enriched fractions = 86
• 5. Proliferation of NK cells = 86
• Cultures of human NK cells = 86
• 6. Natural killer cell cytotoxicity assays = 87
• Human NK cell activity = 87
• Murine natural killer (NK) cell assay = 88
• References = 89
• 6. Isolation and characterization of mononuclear phagocytes / L. J. Partridge ; I. Dransfield = 9l
• 1. Introduction = 91
• 2. Reagents and equipment required for studying mononuclear phagocyte = 91
• 3. Isolation of mononuclear phagocytes = 92
• Density-gradient techniques = 92
• Separation of mononuclear phagocytes by adherence = 94
• Counterflow centrifugal clutiation (CC) = 97
• Immunomagnetic isolation of monocytes = 98
• Isolation of mononuclear phagocytes from solid tumours = 99
• 4. Monocyte subsets = 99
• 5. Ideiltification of mononuclear phagocytes = 100
• Histochcmical staining = 100
• Monoclonal antibody staning = 102
• 6. Functional characterization of mononuclear phagocytes = 108
• Tumour cell killing = 109
• Production of reactive oxygen intermediates = 112
• Chemotaxis = 113
• Phagocytosis = 114
• Antigen presentation = l16
• Cytokine production = 116
• References = 116
• 7. T cells : proliferative and cytotoxic responses / R. Adrian Robins = 119
• 1. Introduction = 119
• 2. Media for lymphocyte preparation and assay = 119
• Salt base, buffer, and nutrients = 119
• Serum or protein supplements = 120
• 3. Dissociation and separation of lymphocytes = 120
• Sources for lymphocyte isolation = 121
• 4. Purification of lymphocytes and T cells = 123
• Density gradients = 123
• Magnetic beads = 124
• 5. Cryopreservation of lymphocytes = 124
• 6. Proliferation assays = 125
• 7. Cytotoxicity assays = 127
• $$\ ^{51}$$Cr release = 127
• Post-label cytotoxicity assays = 128
• References = 129
• 8. Cloning and propagation of human Tlymphocytes / Graham Pawelec = 131
• 1. Introduction = 131
• 2. Tissue-culture medium and growth factors = 132
• 3. Source of cells for cloning, and their activation = 134
• T-cell source = l34
• Activation = 134
• 4. Cloning and propagation of human T lymphocytes = l36
• 5. Cryopreservation = 137
• 6. Assurance of monoclonality = 138
• Acknowledgements = 140
• References = 140
• 9. Association of TCR expression with MHC recognition / Susan L. Hand ; Bruce Lee Hall ; Olivera J. Finn = 143
• 1. Introduction = 143
• 2. Growth of human alloreactive T-cell cultures = 145
• Renal allograft-derived T-cell cultures = 145
• Mixed lymphocyte cultures = 145
• 3. Preparation of total cellular RNA from T-cell cultures = 146
• 4. Synthesis of cDNA from total celluler RNA = 147
• 5. Vβ specific, semiquantitative PCR analysis = 148
• 6. Gel electrophoresis and quantitation of PCR products = 151
• 7. Practical and theoretical considerations of the technique = 152
• Primer selection = 153
• Reaction conditions = 154
• Precision and accuracy = 155
• 8. Representativeness of the message pool = 156
• Acknowledgements = 157
• References = 157
• 10. Cytokines : identification and measurement of gene activation / F. S. di Giovine ; S. Stones ; D. Wojtacha ; G. W. Duff = 159
• 1. Introduction = 159
• 2. Measurement of steady-state mRNA levels = 160
• Northern analysis = 160
• Stot／dot-blot analysis = 168
• Nuclcase protection assay = 168
• Reverse-transcription PCR analysis (RTPCR) = 170
• In sim hybridization = 172
• 3. Transcription rate analysis (nuclear run-on assay) = 174
• Preparation of nuclei = 174
• Labelling of nassent RNA : preparation of probe = 175
• Membrane hybridization = 176
• 4. mRNA transport into cytoplasm = 176
• 5. Cytoplasnlic mRNA half-life = 177
• Ackllowledgements = 177
• References = 178
• 11. Cytokine bioassay / F. P. Winstanley = 179
• 1. Introduction = 179
• 2. Cell proliferation bioassays = 181
• 3. Cytotoxicity bioassays = 183
• 4. Enzyme release bioassays = 185
• 5. Data analysis = 186
• Raw data = 186
• Straightening the curve = 187
• Non-linear fitting of raw response data = 188
• 6. Standards and quality control = l89
• Acknowledgements = 190
• References = 190
• 12. Strategies for the production of human monoclonal antibodies / M. Kurpisz ; G. Wilson = 191
• 1. Introduction = 191
• 2. EBV infection of human B lymphocytes : a practical approach = 193
• Sourcc and preparation of B lymphocytes = 193
• Source of EBV = 196
• In vitro culture systems to improve effciency of EBV transformation = 196
• EBV transformation of B lymphocytes = 200
• Screening for immunoglobulin production = 201
• Maintenance of EBV-transformed cell-lines = 202
• Limiting cell dilution = 204
• Fusion of EBV-transformed cell-lincs = 205
• 3. Clinical use of antibody from EBV-transformed cell-lines = 205
• 4. Conclusions = 205
• References = 206
• 13. Radiolabelling antibodies for imaging and targeting / M. V. Pimm ; S. J. Gribben = 209
• 1. Introduction = 209
• Choice of radioisotopes for antibody labelling = 210
• 2. Handling and measurement of radioisotopes = 211
• Legal responsibilities and safety in handling radioactive materials = 211
• Detection and measurement of gamma-emitting isotopes = 212
• Specific activity of radiolabelled preparations = 212
• 3. Labelling with radioiodine = 213
• Oxidative methods of radioiodination = 213
• Non-oxidative labelling = 216
• Causes of poor labelling = 216
• Detection of free radioiodine in labelled preparations = 217
• 4. Labelling with radioindium = 217
• Causes of poor labelling = 220
• Detection of free radioindium in preparations = 220
• 5. Labelling with technetium = 220
• Causes of poor labelling = 222
• Detection of free $$\ ^{99m}$$Tc in preparations = 222
• 6. Materials and suppliers = 222
• Acknowledgement = 223
• References = 223
• 14. Preparation of multispeciflc F$$\{(ab)}_2$$ and F$${(ab)}_3 $$antibody derivatives / Martin J. Glennie ; Alison L. Tutt ; John Greenman = 225
• 1. Introduction = 225
• 2. Buffer used in the preparation of multispecific derivatives = 226
• 3. Chromatography equipment = 228
• 4. Preparation of mouse lgG and its F$$\{(ab)}_2$$ fragments = 230
• Preparation of mouse lgG = 230
• Preparation of F$$\{(ab)}_2$$ = 231
• 5. F$$\{(ab)}_2$$ and F$$\{(ab)}_3$$ multispecific andtibody derivatives = 233
• Background = 233
• Preparation of multispecific derivatives = 237
• Analysis of antibody fragments on HPLC = 240
• Checking and removing contaminating Fc$$\gamma $$ = 242
• References = 243
• 15. Construction and expression of Ig fusion proteins / Martha S. Hayden ; H. Perry Fell ; Nicholas F. Landolfi = 245
• 1.Introduction = 245
• 2. Design and construction of an immunoligand = 248
• 3. Expression = 248
• Transient mammalian expression of fusion proteins = 248
• Transfection for stable expression = 251
• 4. Detection and characterization of immunoligands = 252
• ELISA = 252
• Biochemical characterization by immunoprecipitation = 253
• Biochemical characterization by Western immunoprecipitation = 254
• Purification of immunoligands by affinity chromatography = 255
• Analysis of the functional activity of the immunoligand = 256
• 5. Antibody fusions = 257
• 6. Conclusion = 260
• References = 260
• 16. Re-shaping human monoclonal antibodies for in vivo diagnosis and therapy / M.E. Verhoeyen = 263
• 1. Introduction = 263
• 2. General strategy for re-shaping human antibodies = 264
• 3. cDNA cloning and sequence determination of antibody variable regions = 265
• 4. Choosing human framework regions for CDR grafting = 266
• 5. Grafting of rodent binding sites on to human framework regions = 268
• CDR grafting by site-directied mutagenesis with large oligonucleotides = 268
• CDR grafting by de novo gene synthesis = 272
• 6. Assembly of re-shaped human V-region genes in functional expression vectors = 273
• 7. Expression of re-shaped human antibody genes in myeloma cells = 273
• 8. Characterization of transfectomas = 275
• References = 277
• 17. Single-chain Fvs / Marc Whitlow ; David Filpula = 279
• 1. Introduction = 279
• 2. Design of the linker between the $$V_L $$ and $$V_H $$ domains = 290
• 3. Genetic construction of a single-chain Fv = 281
• Genetic construction of a single-chain Fv from hybridoma cell-lines = 281
• Genetic construction of a single-chain Fv from combinatiorial libraries = 284
• Expression of a single-chain Fv = 285
• 4. Fermentation and purification of a single-chain Fv = 287
• 5. Design of single-chain Fv fusion proteins = 290
• References = 290
• 18. Engineering cytokine-secreting tumour Cells / Poulam M. Patel ; Claudia L. Flemming ; Suzanne A. Eccles ; Mary K. L. Collins = 293
• 1. Introduction = 293
• 2. Vectors for cytokine gene expression = 294
• 3. Packaging cells = 295
• Care of packaging cells = 296
• Transfecting packaging cells = 296
• Titration of virus production = 298
• Assaying for helper virus = 299
• 4. Cytokine-secreting tumours = 299
• Tumourigenicity assays = 300
• Asscssing cytokine secretion of explanted tumours = 301
• References = 303
• 19. Ex vivo activation of effector cells / K. E. Platts = 305
• 1. Introduction = 305
• 2. Isolation of effector populations = 308
• Peripheral blood mononuclear cells (PBMC) = 308
• Tumour-infiltrating lymphocytes (TIL) = 309
• 3. Separation techniques to isolate enriched or purified populations of effector cells = 310
• 4. Activation of effector populations = 310
• Gcneration of IL-2-activatcd human peripheral blood mononuclear cells = 311
• Generation of IL-2-activated human TIL = 312
• Modifications to thc procedures for the generation of human effector cell populations = 313
• 5. Assessment of activation = 318
• Cytotoxicity assays = 318
• Proliferation = 318
• Cytokine production = 318
• Cytokine gene expression = 318
• Acknowledgement = 319
• References = 319
• 20. Genetic immortalization of human lymphocytes using retroviral vectors / Chris Darnbrough = 321
• 1. Introduction = 321
• 2. Immortalization mechanisms = 322
• Oncogenes = 322
• Autocrine loops = 323
• Imponderables = 324
• Immortalization strategies = 324
• 3. Introducing genes = 324
• Retroviral vectors = 325
• Retroviral vector systems = 325
• Transfection of packaging cells = 329
• HIV-based vectors = 335
• 4. Primary cells = 335
• Sources = 335
• The cell cycle = 336
• Stimulation of primary lymphocytes = 337
• Infection of primary cells = 338
• 5. Conclusion = 341
• Acknowledgements = 341
• References = 342
• 21. Modifying the specificity of T cells using chimeric lg／TCR genes / Zelig Eshhar ; Gideon Gross ; Jonathan Treisman = 345
• 1. Introduction = 345
• 2. Design and construrtion of the cTCR genes = 349
• Antibody selection = 349
• The chimeric gene construct = 350
• Expression vector systcms = 350
• Cloning of cTCR into mammalian expression vectors = 351
• 3. Transfection and selection of cells expressing the cTCR genes = 355
• Choosing cells for transfection = 355
• Sequential transfcction versus co-transfection = 356
• Methods of DNA transfection = 356
• Selection of cells using antibiotic resistance = 358
• 4. Selection of cells expressing the cTCR = 360
• Molccular analysis of transfectants = 360
• Functional analysis of transfectants = 364
• Acknowledgement = 367
• References = 367
• 22. The use of pre-clinical rodent models for cancer immunotherapy / Robert H. Wiltrout = 369
• 1. Introduction = 369
• 2. General types of rodent tumour models = 369
• Transplantable primary tumours = 370
• Transpl metastatic tumours = 371
• Primary autochthonous tumours = 372
• 3. Application and description of a transplantable murine renal cancer model = 373
• Intrarenal Renca as a model for treatment of a primary tumour = 373
• The use of Renca for the formation of experimental pulmonary metastases = 376
• The use of Renca for the formation of experimental hepatic metastases = 377
• Summary of pre-clinicla immunotherapy results obtained using the Tenca tumour model = 378
• Relationship between clinical results and pre-chenical predictions = 379
• 4. General survey of widely used pre-chanical rodent models = 380
• 5. Implications of biological variability that exists between rodent tumour models = 380
• 6. Summary = 382
• References = 382
• 23. Use of the tumour spheroid model in immunotherapy / Irma E. Garcia de Palazzo ; Michele Holmes ; Katherine Alpaugh ; Louis M. Weiner = 385
• 1. Introduction = 385
• 2. Methods and experimental design = 386
• MHTS generation and growth = 386
• Preparation of IL-2-activated lymphocytes = 387
• Antibodies = 397
• Co-culture of MHTS with LAK cells and antibodies = 388
• Histology and immunohistochemistry = 388
• 3. Results = 389
• Morphology and differentiation characteristics of SW948 = 389
• Histological and immunohistochemical charactcrization of LAK cell infiltration of MHTS = 390
• 4. Concluding remarks = 395
• References = 396
• 24. Characterization of metastatic tumour cells / U. P. Thorgeirsson ; A. R. Mackay = 399
• 1. Introduction = 399
• 2. Animal models = 399
• 3. In vivo metastasis assays = 401
• Tumour cell preparation = 401
• Routes of injection = 401
• 4. In vitro assay to study the metastatic phenotype = 404
• Tumour invasion assays = 404
• Amnion invasion assay = 405
• 5. Proteolytic activity = 406
• 6. Tumour cell surface = 409
• References = 409
• Index = 413