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Over the past 15 yr, the use of transgenic mice has led to significant advances in our understanding of immunological tolerance. In a normal repertoire the number of B cells with a single antigen receptor specificity is very small, making the study of their fate difficult. In contrast, animals that carry transgenes encoding rearranged immunoglobulin genes generate large numbers of B cells that, by the process of allelic exclusion, have an identical specificity. Exploitation of this effect has enabled the mechanisms involved in B-cell tolerance to be explored in some detail. In this review we use the hen egg lysozyme (HEL) model system to illustrate the generation and preparation of a transgene. In our example, we describe the generation of mice expressing HEL as a systemic, intracellular, membrane-bound self-antigen. The same principles and methods apply to immunoglobulin transgenes. We briefly discuss the techniques that could be used to explore mechanisms of tolerance to systemic intracellular antigens in these mice.

Original publication

DOI

10.1385/1-59259-796-3:239

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2004

Volume

271

Pages

239 - 260

Keywords

Animals, B-Lymphocytes, Blotting, Southern, Female, Immune Tolerance, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Microinjections, Models, Immunological, Muramidase, Receptors, Antigen, B-Cell, Transgenes, Tumor Cells, Cultured