Lymphocytes: a practical approach
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
Oxford
Oxford Univ. Pr.
2000
|
| Ausgabe: | 2. ed. |
| Schriftenreihe: | The practical approach series
220 |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XXIII, 352 S. graph. Darst. |
| ISBN: | 0199638179 0199638160 |
Internformat
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| 245 | 1 | 0 | |a Lymphocytes |b a practical approach |c ed. by Sarah L. Rowland-Jones ... |
| 250 | |a 2. ed. | ||
| 264 | 1 | |a Oxford |b Oxford Univ. Pr. |c 2000 | |
| 300 | |a XXIII, 352 S. |b graph. Darst. | ||
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| 490 | 1 | |a The practical approach series |v 220 | |
| 650 | 4 | |a Immunologic Techniques |v Laboratory Manuals | |
| 650 | 4 | |a Lymphocytes |v Laboratory manuals | |
| 650 | 4 | |a Lymphocytes |x immunology |v Laboratory Manuals | |
| 700 | 1 | |a Rowland-Jones, Sarah L. |e Sonstige |4 oth | |
| 830 | 0 | |a The practical approach series |v 220 |w (DE-604)BV007921321 |9 220 | |
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Datensatz im Suchindex
| _version_ | 1804127775012945920 |
|---|---|
| adam_text | Contents
List of Contributors xvii
Abbreviations xix
1. Preparation of lymphocytes and identification
of lymphocyte subpopulations 1
Magdalena Plebanski
1. Introduction 1
2. Sources of lymphoid and professional antigen presenting
cells 1
3. Methods of purification 3
Lymphoid organs 3
Body fluids 10
4. Principles of lymphocyte handling and culture 14
Short and long term storage (freezing) 14
Principles of long and short term culture 16
5. Methods of subfractionation for lymphocytes and
antigen presenting cells 18
Physical properties 19
Cell surface markers 20
Acknowledgement 26
2. Immunohistochemistry of lymphoid organs 27
Simon C. Biddolph and Kevin C. Gatter
1. Introduction . 27
2. Preparation of lymphoid cells and tissues for
immunohistochemistry 27
Slide adhesive 27
Choice of preparation 28
Transport of specimen 29
Cell smear and cytocentrifuge preparations 29
Cell imprints ( dabs or touch preparations ) 31
Frozen sections ( cryostat sections ) 31
Paraffin sections 32
3. Immunohistochemical staining 39
Immunohistochemical methods 39
Contents
Labels 41
Considerations in the choice of method and label 45
Multiple antibody methods 49
Quality control 52
The distribution of cell types in lymphoid tissue 53
Acknowledgements 53
References 53
3. Viral transformation of lymphocytes 55
Edgar Meinl and Helmut Fickenscher
1. Introduction 55
2. Transformation of T lymphocytes 56
Growth transformation of human T cells by herpesvirus saimiri 56
Immortalization of human T cells by human T cell leukaemia
virus type 1 °
3. B cell immortalization 69
EB V transf ormation of human B cells
Transformation of macaque B cells with herpesvirus papio 70
Acknowledgements 71
References 71
4. B and T cell hybridomas 75
Judy M. Bastin
1. Introduction 75
2. Methods for generating B and T cell hybridomas and the
principles of hybridoma culture 75
Tissue culture 76
Materials 76
Selection procedures 77
Immunizations 77
Fusion partners 79
Fusion protocol 79
Growth of hybridomas 81
Screening assays 81
Cloning hybridomas 82
Cryopreserving and thawing cells 83
Large scale production of hybridomas 84
Purification 84
Human hybridomas 85
Antibody production by chemical and genetic engineering 86
x
Contents
Disadvantages of hybridomas 87
Conclusions 87
3. Applications of hybridoma technology 88
B cell hybridomas 88
T cell hybridomas 90
References 91
5. Murine T cell culture 95
Kingston H. G. Mills
1. Introduction 95
T cell subtypes and their role in protective immunity 95
Strategies for the induction of distinct T cell subtypes 96
Techniques for detecting T cell responses 98
2. Tissue culture conditions, growth factors, and cell viability 99
Culture medium and serum 99
Cytokines and growth factors for culturing T cells 101
Viable cell count 102
3. Preparation of T cells and APCs 103
Preparation of cells from lymphoid and non lymphoid organs 103
Purification of mononuclear cells and removal of dead cells 106
Purification of T cells, B cells, and T cell subpopulations 107
Preparation of macrophages and APCs 110
4. Generation and detection of antigen specific CD4+ T cells 111
In vivo priming of CD4+ T cells 111
Generation of CD4+ T cell lines and clones 112
Antigen specific T cell proliferation 115
Detection of Thl/ThO/Th2 responses by cytokine production 119
Assay of helper function of T cells for antibody production by B cells 124
5. Generation and detection of antigen specific CD8+ CTLs 127
In vivo priming of CD8+CTLs 127
Generation of CD8+ T cell lines and clones 127
Cytotoxic T cell assay 128
6. MHC restriction analysis 130
Acknowledgements 131
References 132
6. Human CD4 culture 135
Anna Vyakamam, B. Vyas, M. Vukmanovic Stejic,
P. Gorak Stolinska, D. Wallace, A. Noble, and D. M. Kemeny
1. Introduction 135
xi
Contents
2. General apparatus, media, and reagents for human CD4 culture 136
Apparatus 136
Media 137
Reagents 137
3. The isolation and culture of CD4 T cell subsets from peripheral
blood 137
Principle 140
The generation of CD4+ Thl, Th2, and ThO effectors 140
Generation of antigen specific and random CD4 clones 142
4. Assays for CD4 function 147
Principle of proliferation assay 147
Activation induced cell death 149
Measuring cytokine production in culture supernatants by ELISA 151
Enumerating cytokihe producing cells by flow cytometry:
intracytoplasmic cytokine staining 153
References 158
7. Human cytotoxic T lymphocyte (GTL) studies i6i
Sarah L. Rowland Jones
1. Introduction 161
2. General principles of CTL culture 162
Tissue culture reagents and conditions 162
What persuades memory CTLs to grow in vitro? 163
3. Generation of antiviral CTL 163
Influenza specific CTLs 164
HIV specific CTLs 165
Using peptides to stimulate CTLs 166
Generating virus specific CTL clones 167
4. Measuring CTL activity 169
The CTL lysis assay 169
Measuring the antigen specific release of cytokines by CTLs 172
How do different methods of quantifying CTLs compare? 175
5. Conclusions 176
References 176
8. Limiting dilution analysis for the quantitation
of antigen specific T cells 179
Andrew Carmichael
1. Theoretical considerations 179
Quantitation at the clonal level using limiting dilution analysis (LDA) 179
The Poisson distribution 180
xii
Contents
Advantages of LDA in addition to quantitation 183
Limitations of LDA 183
2. Methodology 183
Responder cells 184
Stimulator cells 187
3. Calculation and interpretation of results 190
Calculation of cytotoxic activity in individual microcultures 190
Classification of positive and negative wells 190
Estimation of precursor frequency 191
Interpretation 194
References 194
9. HLA peptide tetrameric complexes 197
Graham S. Ogg
1. Introduction 197
2. Cloning of modified HLA heavy chain 198
3. Protein expression 199
Principle 199
Comments 200
4. Inclusion body purification 200
Principle 200
5. Refolding by dilution 201
Principle 201
Comments 202
6. Enzymatic biotinylation 202
Principle 202
Comments 203
7. Purification of refolded protein 203
Principle 203
8. ELISA 204
Principle 204
Comments 206
9. Formation of tetramers 206
Principle 206
Comments 206
10. Flow cytometry 207
Principle 207
Comments 207
11. Conclusions 207
References 208
xiii
Contents
10. Expansion of human cytotoxic T lymphocytes
for immunotherapy 209
Rusung Tan
1. Introduction 209
Adoptive transfer of CTLs in humans 210
2. Peptide recognition by CTLs 211
Peptide binding motifs 211
3. Establishing CTL lines 213
Establishing CTL clones 213
Expansion of CTL clones 215
Ensuring the safety of CTL infusions 217
References 218
11. HLA typing methods 221
Pete Krausa and Michael Browning
1. Introduction 221
2. The HLA system 223
Polymorphism in relation to structure and function 224
Nomenclature 226
HLA in populations 227
3. Methods of HLA typing 230
4. Serological HLA typing 230
Preparation of components 232
The micro lymphocytotoxicity assay 237
Discussion on serology 238
5. Molecular biological approaches to tissue typing 239
6. The PCR reaction 240
Reagent composition 240
The PCR 242
7. Detection by oligonucleotide probing (PCR SSOP) 243
PCR SSOP protocols 245
PCR SSOP discussion 248
8. HLA typing by PCR SSP 249
Five locus determination PCR SSP—phototyping 249
High resolution PCR SSP typing 256
PCR SSP discussion 257
9. Sequence based typing 258
SBT strategies 259
HLA SBT protocol 261
xiv
Contents
j 10. Discussion 264
: Key references 267
Commercial suppliers 267
References 268
12. Biochemical characterization of lymphocyte
surface antigens 273
Antony Symons and Marion H. Brown
1. Introduction 273
2. Immunoprecipitation 274
Introduction 274
Labelling cellular antigens 274
Solubilization of membrane proteins 280
Immunoprecipitation 281
3. SDS polyacrylamide gel electrophoresis (PAGE) 285
Introduction 285
Detection of proteins 288
4. Immunoblotting 289
Introduction 289
Preparation of protein samples 290
Blotting proteins 291
Detection of antigen 294
5. Strategies for the identification of new molecules 296
Affinity chromatography 296
Ligand hunting 297
References 297
13. Measurement of cells undergoing apoptosis 299
Y. Furukawa and C. R. M. Bangham
1. Introduction 299
Definition of apoptosis 299
Importance of apoptosis in lymphocyte biology 299
Recognition of apoptotic cells by phagocytes 300
Recent advances in our understanding of apoptosis 300
2. Methods available to assay apoptosis 303
DNA laddering 305
Analysis of light scatter by flow cytometry (size and granularity) 305
xv
Contents
Terminal deoxynucleotidyl transferase mediated dUTP nick
end labelling (TUNEL) 305
Annexin V 308
CMXRos and related dyes 309
References 313
14. Thymic organ culture 315
John J. T. Owen and Deirdre E. J. McLoughlin
1. Introduction 315
2. Microdissection and organ culture of the murine fetal
thymus 316
The production of alymphoid thymus lobes 321
3. Newborn thymic slice culture 322
4. Reaggregate thymus organ culture 324
References 329
Appendices
Al List of suppliers 331
A2 Commonly used lymphoma lines 337
Edgar Meinl and Helmut Fickenscher
Index 343
xvi
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| indexdate | 2024-07-09T18:38:42Z |
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| publisher | Oxford Univ. Pr. |
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| series | The practical approach series |
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| spelling | Lymphocytes a practical approach ed. by Sarah L. Rowland-Jones ... 2. ed. Oxford Oxford Univ. Pr. 2000 XXIII, 352 S. graph. Darst. txt rdacontent n rdamedia nc rdacarrier The practical approach series 220 Immunologic Techniques Laboratory Manuals Lymphocytes Laboratory manuals Lymphocytes immunology Laboratory Manuals Rowland-Jones, Sarah L. Sonstige oth The practical approach series 220 (DE-604)BV007921321 220 HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008909604&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Lymphocytes a practical approach The practical approach series Immunologic Techniques Laboratory Manuals Lymphocytes Laboratory manuals Lymphocytes immunology Laboratory Manuals |
| title | Lymphocytes a practical approach |
| title_auth | Lymphocytes a practical approach |
| title_exact_search | Lymphocytes a practical approach |
| title_full | Lymphocytes a practical approach ed. by Sarah L. Rowland-Jones ... |
| title_fullStr | Lymphocytes a practical approach ed. by Sarah L. Rowland-Jones ... |
| title_full_unstemmed | Lymphocytes a practical approach ed. by Sarah L. Rowland-Jones ... |
| title_short | Lymphocytes |
| title_sort | lymphocytes a practical approach |
| title_sub | a practical approach |
| topic | Immunologic Techniques Laboratory Manuals Lymphocytes Laboratory manuals Lymphocytes immunology Laboratory Manuals |
| topic_facet | Immunologic Techniques Laboratory Manuals Lymphocytes Laboratory manuals Lymphocytes immunology Laboratory Manuals |
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