Immunology of Infection, 2nd Edition, edited by two leading experts in the field, presents the most appropriate up-to-date experimental approaches in the detail required for modern microbiological research. Focusing on the methods most useful for the Microbiologist interested in analysing host-pathogen relationships, this volume will be essential reading for all researchers working in microbiology, immunology, virology, mycology and parasitology. This new edition of Immunology of Infection provides ready-to-use "recipes", and the latest emerging techniques as well as novel approaches to the tried and tested, established methods included in the successful first edition. Methods in Microbiologyis the most prestigious series devoted to techniques and methodology in the field. Established for over 30 years, Methods in Microbiology will continue to provide you with tried and tested, cutting edge protocols to directly benefit your research.
Key Features: * Includes techniques for genome-wide expression profiling of both the pathogen and host and of the host response to infection * Cytometric analysis of cytokine secretion by immune cells * Describes tetramer technology for the quantitative analysis of antigen specific T cell responses * analysis of host cells and pathogens involved in the host-microbe interplay * Covers techniques useful for the analysis of human and murine systems * Includes techniques for the prediction and determination of MHC ligands and T cell epitopes * Covers the fundamentals and practice of DNA vaccines * Describes methods for the isolation and propagation of human dendritic cells
Contributors Preface to the 2nd edition Preface to the 1st edition Introduction The immune response to infectious agents General Methods Phenotyping and separation of leukocyte populations based on affinity labelling. Cytometric cytokine secretion assay: detection and isolation of cytokine secreting T cells. Measurement of cellular proliferation. Use of bioinformatics to predict MHC ligands and T-cell epitopes: application to epitope driven vaccine design. The use of tetramers in the quantitative analysis of T cell responses. Using microarrays for studying the host transcriptional response to microbial infection. Genome-wide expression profiling of intracelllular bacteria: the interaction of Mycobacterium tuberculosis with macrophages. Murine Models Management of immuno-compromised and infected animals. In Vitro Analysis Isolation and preparation of lymphocytes from infected animals for in vitro analysis. Isolation and development of murine T-lymphocyte hybridomas and clonal cell lines. Killer cell assays. Quantitation of T Cell Cytokine Responses by ELISA, ELISPOT, Flow Cytometry and Reverse Transcriptase-PCR Methods. Isolation of and measuring the function of professional phagocytes: murine macrophages. In vivo Analysis The immune response in mice challenged with bacterial infections. Measuring immune responses in vivo. Specific Models Murine model of tuberculosis. The leishmaniasis model. Animal models: murine cytomegalovirus. DNA vaccines: fundamentals and practice. Preparation and use of adjuvants. Human Systems Isolation and propagation of human dendritic cells. Isolation of T cells and establishment of T-cell lines and clones. Growth-transformation of human T cells. Generation and characterisation of human killer cells. Measuring human cytokine responses. Measuring immune responses in situ: immunofluorescent and immunoenzymatic techniques. Isolation, characterisation and cultivation of human monocytes and macrophages.