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Table of Contents:

Contributors
Preface to the First Edition
Preface to the Second Edition
Introduction: The immune response to infectious agents 1
I General Methods
1 Phenotyping and separation of leukocyte populations based on affinity labelling 23
2 Cytometric cytokine secretion assay: detection and isolation of cytokine-secreting T cells 59
3 Measurement of cellular proliferation 77
4 Use of bioinformatics to predict MHC ligands and T-cell epitopes: application to epitope-driven vaccine design 99
5 The use of tetramers in the quantitative analysis of T-cell responses 125
6 Using microarrays for studying the host transcriptional response to microbial infection 157
7 Genome-wide expression profiling of intracellular bacteria: the interaction of Mycobacterium tuberculosis with macrophages 169
II Murine Models
1 Management of immunocompromised and infected animals 183
2 In vitro analysis
2.1 Isolation and preparation of lymphocytes from infected animals for in vitro analysis 233
2.2 Isolation and development of murine T-lymphocyte hybridomas and clonal cell lines 249
2.3 Killer cell assays 281
2.4 Quantitation of T-cell cytokine responses by ELISA, ELISPOT, flow cytometry and reverse transcriptase-PCR methods 301
2.5 Isolation of and measuring the function of professional phagocytes: murine macrophages 331
3 In vivo analysis
3.1 The immune response in mice challenged with bacterial infections 359
3.2 Measuring immune responses in vivo 403
4 Specific Models
4.1 Murine models of tuberculosis 433
4.2 The leishmaniasis model 463
4.3 Animal models: murine cytomegalovirus 493
4.4 DNA vaccines: fundamentals and practice 527
4.5 Preparation and use of adjuvants 551
III Human Systems
1 Isolation and propagation of human dendritic cells 591
2 Isolation of T cells and establishment of T-cell lines and clones 621
3 Growth transformation of human T cells 657
4 Generation and characterization of human killer cells 693
5 Measuring human cytokine responses 707
6 Measuring immune responses in situ: immunofluorescent and immunoenzymatic techniques 751
7 Isolation, characterization and cultivation of human monocytes and macrophages 767
Index 787